2d-and-3d-matrices-to-study-linear-invadosome-formation-and-activity

2D and 3D Matrices to Study Linear Invadosome Formation and Activity

Cell adhesion, migration, and invasion are involved in many physiological and pathological processes. For example, during metastasis formation, tumor ...

Tisch Cancer Institute

We report a highly specific, sensitive, and robust method for analyzing fluorescence resonance energy transfer (FRET) based on spectral laser scanning confocal microscopy imaging. The lambda FRET (lambdaFRET) algorithm comprises imaging of a FRET sample at multiple emission wavelengths rendering a FRET spectrum, which is separated into its donor and acceptor components to obtain a pixel-based calculation of FRET efficiency. The method uses a novel off-line precalibration procedure for spectral bleed-through correction based on the acquisition of reference reflection images, which simplifies the method and reduces variability. LambdaFRET method was validated using structurally characterized FRET standards with variable linker lengths and stoichiometries designed for this purpose. LambdaFRET performed better than other well-established methods, such as acceptor photobleaching and sensitized emission-based methods, in terms of specificity, reproducibility, and sensitivity to distance variations. Moreover, lambdaFRET analysis was unaffected by high fluorochrome spectral overlap and cellular autofluorescence. The lambdaFRET method demonstrated outstanding performance in intra- and intermolecular FRET analysis in both fixed and live cell imaging studies.

Novel lambda FRET spectral confocal microscopy imaging method.

Invadopodia &#x662F;&#x57FA;&#x8D28;&#x964D;&#x89E3;&#x819C;&#x7A81;&#x8D77;&#x4E2D;&#x6D78;&#x6DA6;&#x764C;&#x7EC6;&#x80DE;&#x4E2D;&#x8C03;&#x8282;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x805A;&#x5408;&#x7684;&#x86CB;&#x767D;&#x8D28;&#x4E30;&#x5BCC;&#x3002;&#x89C4;&#x7BA1;&#x5F00;&#x5173;&#x542F;&#x52A8;&#x5728; invadopodia &#x4E2D;&#x7684;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x805A;&#x5408;&#x7684;&#x9700;&#x8981;&#x78F7;&#x9178;&#x5316;&#x7684;&#x916A;&#x6C28;&#x9178;&#x6B8B;&#x57FA; 421&#x3001; 466&#x3001; &#x548C; 482 &#x4E0A;&#x76AE;&#x5C42;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x5B83;&#x662F;&#x672A;&#x77E5;&#x7684;&#x8FD9;&#x4E9B;&#x76AE;&#x5C42;&#x916A;&#x6C28;&#x9178;&#x78F7;&#x9178;&#x5316;&#x7AD9;&#x70B9;&#x63A7;&#x5236;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x7684;&#x805A;&#x5408;&#x3002;&#x6211;&#x4EEC;&#x8C03;&#x67E5;&#x4E2A;&#x522B;&#x916A;&#x6C28;&#x9178;&#x78F7;&#x9178;&#x5316;&#x7F51;&#x7AD9; (421&#x3001; 466&#x3001; &#x548C; 482) &#x7684;&#x8D21;&#x732E;&#x4E0A;&#x76AE;&#x5C42;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x805A;&#x5408; invadopodia &#x5728;&#x89C4;&#x7BA1;&#x3002;&#x6211;&#x4EEC;&#x63D0;&#x4F9B;&#x8BC1;&#x636E;&#x8868;&#x660E;&#xFF0C;&#x78F7;&#x9178;&#x5316;&#x7684;&#x916A;&#x6C28;&#x9178; 421 &#x548C; 466&#xFF0C;&#x4F46;&#x4E0D;&#x662F; 482&#xFF0C;&#x662F;&#x9700;&#x8981;&#x5728; invadopodia &#x514D;&#x8D39;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x523A;&#x4E24;&#x7AEF;&#x7684;&#x4E00;&#x4EE3;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x91CD;&#x8981;&#x7684;&#x662F;&#x901A;&#x8FC7;&#x5176; SH2 &#x57DF;&#xFF0C;&#x7528;&#x4E8E;&#x76F4;&#x63A5;&#x7ED1;&#x5B9A;&#x5230;&#x76AE;&#x5C42;&#x5728;&#x8BD5;&#x7BA1;&#x91CC;&#xFF0C;Nck1 &#x7684;&#x548C;&#x4E3A; Nck1 &#x548C; invadopodia &#x7684;&#x76AE;&#x5C42;&#x4E4B;&#x95F4;&#x7684;&#x76F8;&#x4E92;&#x4F5C;&#x7528;&#x97F3;&#x54C1; invadopodia Nck1 &#x62DB;&#x52DF;&#x8FD9;&#x4E9B;&#x76F8;&#x540C;&#x7684; phosphotyrosines&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x5728;&#x7EC6;&#x80DE;&#x8868;&#x8FBE;&#x4E00;&#x79CD;&#x7A81;&#x53D8;&#x916A;&#x6C28;&#x9178; 421 &#x6216; 466 &#x5927;&#x5927;&#x53D7;&#x5230;&#x6291;&#x5236;&#x77E9;&#x9635;&#x4F9D;&#x8D56;&#x86CB;&#x767D;&#x8D28;&#x964D;&#x89E3;&#x7684;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x4FB5;&#x88AD;&#x3002;&#x5728;&#x4E00;&#x8D77;&#xFF0C;&#x8FD9;&#x4E9B;&#x7ED3;&#x679C;&#x4F5C;&#x4E3A;&#x8C03;&#x8282; invadopodia Nck1 &#x4F9D;&#x8D56;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x805A;&#x5408;&#x7684;&#x5173;&#x952E;&#x6B8B;&#x6E23;&#x8BC6;&#x522B;&#x78F7;&#x9178;&#x5316;&#x916A;&#x6C28;&#x9178; 421 &#x548C; 466 &#x4E0A;&#x76AE;&#x5C42;&#x548C;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x5165;&#x4FB5;&#xFF0C;&#x5E76;&#x5EFA;&#x8BAE;&#x4E13;&#x95E8;&#x963B;&#x6B62;&#x4EFB;&#x4E00; 421 &#x6216; 466 &#x7684;&#x916A;&#x6C28;&#x9178;&#x78F7;&#x9178;&#x5316;&#x53EF;&#x80FD;&#x4F1A;&#x6709;&#x6548;&#x5730;&#x6291;&#x5236;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x5165;&#x4FB5;&#x4F53;&#x5185;&#x3002;

&#x5177;&#x4F53;&#x7684;&#x916A;&#x6C28;&#x9178;&#x78F7;&#x9178;&#x5316;&#x7AD9;&#x70B9;&#x4E0A;&#x76AE;&#x5C42;&#x89C4;&#x7BA1; invadopodia Nck1 &#x4F9D;&#x8D56;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x805A;&#x5408;&#x3002;

Invadopodia are matrix-degrading membrane protrusions in invasive carcinoma cells enriched in proteins that regulate actin polymerization. The on-off regulatory switch that initiates actin polymerization in invadopodia requires phosphorylation of tyrosine residues 421, 466, and 482 on cortactin. However, it is unknown which of these cortactin tyrosine phosphorylation sites control actin polymerization. We investigated the contribution of individual tyrosine phosphorylation sites (421, 466, and 482) on cortactin to the regulation of actin polymerization in invadopodia. We provide evidence that the phosphorylation of tyrosines 421 and 466, but not 482, is required for the generation of free actin barbed ends in invadopodia. In addition, these same phosphotyrosines are important for Nck1 recruitment to invadopodia via its SH2 domain, for the direct binding of Nck1 to cortactin in vitro, and for the FRET interaction between Nck1 and cortactin in invadopodia. Furthermore, matrix proteolysis-dependent tumor cell invasion is dramatically inhibited in cells expressing a mutation in phosphotyrosine 421 or 466. Together, these results identify phosphorylation of tyrosines 421 and 466 on cortactin as the crucial residues that regulate Nck1-dependent actin polymerization in invadopodia and tumor cell invasion, and suggest that specifically blocking either tyrosine 421 or 466 phosphorylation might be effective at inhibiting tumor cell invasion in vivo.

Specific tyrosine phosphorylation sites on cortactin regulate Nck1-dependent actin polymerization in invadopodia.

Invadopodia sont capables de d&eacute;grader matrice protub&eacute;rances membranaires dans les cellules de carcinome invasif enrichis en prot&eacute;ines qui r&eacute;gulent la polym&eacute;risation de l&#39;actine. Le commutateur marche-arr&ecirc;t r&eacute;glementaire qui initie la polym&eacute;risation de l&#39;actine dans invadopodia exige la phosphorylation des r&eacute;sidus tyrosine 421, 466 et 482 sur cortactin. Toutefois, il est inconnu, lequel de ces sites de phosphorylation de tyrosine cortactin contr&ocirc;ler la polym&eacute;risation de l&#39;actine. Nous avons &eacute;tudi&eacute; la contribution des sites de phosphorylation de tyrosine individuels (421, 466 et 482) sur cortactin pour le r&egrave;glement de la polym&eacute;risation de l&#39;actine dans invadopodia. Nous fournissons la preuve que la phosphorylation des tyrosines 421 et 466, mais pas de 482, est requis pour la g&eacute;n&eacute;ration des extr&eacute;mit&eacute;s de l&#39;actine libre barb&eacute;e d&#39;invadopodia. En outre, ces m&ecirc;mes phosphotyrosines sont importants de recrutement Nck1 invadopodia via son domaine SH2, pour la liaison directe de Nck1 &agrave; cortactin in vitro et pour l&#39;interaction frette entre Nck1 et cortactin dans invadopodia. En outre, invasion des cellules tumorales de prot&eacute;olyse d&eacute;pendante matrice est consid&eacute;rablement inhib&eacute;e dans les cellules exprimant une mutation de la phosphotyrosine 421 ou 466. Ensemble, ces r&eacute;sultats identifient la phosphorylation des tyrosines 421 et 466 sur cortactin comme les r&eacute;sidus cruciaux qui r&eacute;gulent la polym&eacute;risation de l&#39;actine dans invadopodia Nck1-d&eacute;pendante et invasion des cellules tumorales et sugg&egrave;rent que sp&eacute;cifiquement bloquant soit la phosphorylation de tyrosine 421 ou 466 pourrait &ecirc;tre efficace pour inhiber l&#39;invasion des cellules tumorales in vivo.

Sites de phosphorylation de tyrosine sp&eacute;cifique sur cortactin r&eacute;glementent la polym&eacute;risation de l&#39;actine Nck1-d&eacute;pendante dans invadopodia.

Invadopodia sind Matrix-erniedrigende Membran Vorspr&uuml;nge in invasive Karzinom-Zellen angereichert in Proteinen, die Aktin-Polymerisation zu regulieren. Der auf-regulatorische Ausschalter, die Aktin-Polymerisation in Invadopodia initiiert erfordert Phosphorylierung von Tyrosin R&uuml;ckst&auml;nde 421, 466 und 482 auf Cortactin. Es ist jedoch unbekannt, die dieser Cortactin Tyrosin-Phosphorylierung-Sites Aktin-Polymerisation zu steuern. Wir untersuchten den Beitrag einzelner Tyrosin Phosphorylierung Sites (421, 466 und 482) auf Cortactin zur Regulierung der Aktin-Polymerisation in Invadopodia. Wir nachweisen, dass die Phosphorylierung von Tyrosines 421 und 466, aber nicht 482, wird ben&ouml;tigt zur Erzeugung von freien Aktin Stacheldraht endet in Invadopodia. Dar&uuml;ber hinaus sind diese gleichen Phosphotyrosines wichtig f&uuml;r Nck1 Rekrutierung von Invadopodia &uuml;ber seine SH2-Dom&auml;ne, f&uuml;r die direkte Bindung des Nck1 an Cortactin in-vitro- und f&uuml;r die Bund-Interaktion zwischen Nck1 und Cortactin in Invadopodia. Dar&uuml;ber hinaus wird die Matrix Proteolyse-abh&auml;ngige Zelle Tumorinvasion dramatisch in Zellen, die Ausdruck einer Mutation im Phosphotyrosine 421 oder 466 gehemmt. Zusammen diese Ergebnisse ermitteln Phosphorylierung von Tyrosines 421 und 466 auf Cortactin als die entscheidende R&uuml;ckst&auml;nde, die Nck1-abh&auml;ngige Aktin-Polymerisation in Invadopodia Regeln und Tumor Zellen-Einmarsch und Vermutung nahe, dass speziell blockieren entweder 421 oder 466 Tyrosin-Phosphorylierung effektiv bei der Hemmung der Zelle Tumorinvasion in-vivo.

Spezifische Tyrosin-Phosphorylierung-Sites auf Cortactin Regeln Nck1-abh&auml;ngige Aktin-Polymerisation in Invadopodia.

Invadopodia sono degradanti matrix protrusioni di membrana in cellule di carcinoma invasivo arricchite in proteine che regolano la polimerizzazione dell&#39;actina. L&#39;interruttore on-off normativo che avvia la polimerizzazione dell&#39;actina in invadopodia richiede la fosforilazione dei residui di tirosina 421, 466 e 482 su cortactin. Tuttavia, &egrave; ignoto, quale di questi siti di fosforilazione della tirosina cortactin controllare la polimerizzazione dell&#39;actina. Abbiamo studiato il contributo dei siti di fosforilazione della tirosina individuali (421, 466 e 482) su cortactin per la regolazione della polimerizzazione dell&#39;actina in invadopodia. Forniamo la prova che la fosforilazione delle tirosine 421 e 466, ma non 482, &egrave; richiesto per la generazione delle estremit&agrave; libera dell&#39;actina spinato in invadopodia. Inoltre, queste stesse phosphotyrosines sono importanti per l&#39;assunzione di Nck1 a invadopodia attraverso il dominio SH2, per il binding diretto di Nck1 di cortactin in vitro e per il FRET interazione tra Nck1 e cortactin in invadopodia. Inoltre, invasione di matrice proteolisi dipendente dal tumore delle cellule &egrave; drammaticamente inibita nelle cellule che esprimono una mutazione phosphotyrosine 421 o 466. Insieme, questi risultati identificano la fosforilazione delle tirosine 421 e 466 su cortactin come i residui fondamentali che regolano la polimerizzazione dell&#39;actina Nck1-dipendente in invadopodia e tumore invasione di cella e suggeriscono che specificamente bloccando o fosforilazione della tirosina 421 o 466 potrebbe essere efficacia nell&#39;inibire l&#39;invasione di cellule tumorali in vivo.

Siti fosforilazione della tirosina specifico su cortactin regolano la polimerizzazione dell&#39;actina Nck1-dipendente in invadopodia.

Invadopodia &#x306F;&#x30DE;&#x30C8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x5206;&#x89E3;&#x6D78;&#x6F64;&#x764C;&#x7D30;&#x80DE;&#x306E;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x91CD;&#x5408;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA;&#x306E;&#x6FC3;&#x7E2E;&#x306B;&#x304A;&#x3051;&#x308B;&#x819C;&#x7A81;&#x8D77;&#x3002;Invadopodia &#x3067;&#x306E;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x91CD;&#x5408;&#x3092;&#x958B;&#x59CB;&#x3059;&#x308B;&#x30AA;&#x30F3;-&#x30AA;&#x30D5;&#x898F;&#x5236;&#x30B9;&#x30A4;&#x30C3;&#x30C1; &#x30C1;&#x30ED;&#x30B7;&#x30F3;&#x6B8B;&#x57FA; 421, 466, &#x3068; 482 cortactin &#x4E0A;&#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;&#x3002;&#x305F;&#x3060;&#x3057;&#x3001;&#x3053;&#x308C;&#x3089; cortactin &#x30C1;&#x30ED;&#x30B7;&#x30F3;&#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x306E;&#x30B5;&#x30A4;&#x30C8;&#x306E;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x91CD;&#x5408;&#x5236;&#x5FA1;&#x672A;&#x77E5;&#x3067;&#x3059;&#x3002;Cortactin invadopodia &#x306E;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x91CD;&#x5408;&#x306E;&#x898F;&#x5236;&#x306B;&#x500B;&#x3005; &#x306E;&#x30C1;&#x30ED;&#x30B7;&#x30F3; &#x30EA;&#x30F3;&#x9178;&#x5316;&#x30B5;&#x30A4;&#x30C8; &#xFF08;421, 466, &#x3068; 482&#xFF09; &#x306E;&#x5BC4;&#x4E0E;&#x3092;&#x8ABF;&#x67FB;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x6211;&#x3005; &#x304C;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x8A3C;&#x62E0;&#x306F;&#x3001;&#x30EA;&#x30F3;&#x9178;&#x5316; tyrosines &#x306E; 421 &#x3068; 466&#x3001;&#x306A;&#x304F;&#x306A;&#x3044; 482 &#x5FC5;&#x8981;&#x3067;&#x3059; invadopodia &#x306E;&#x7121;&#x6599;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x306E;&#x6709;&#x523A;&#x9244;&#x7DDA;&#x306E;&#x4E21;&#x7AEF;&#x306E;&#x4E16;&#x4EE3;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x540C;&#x3058; phosphotyrosines Nck1 invadopodia Nck1 cortactin in vitro &#x3078;&#x306E;&#x76F4;&#x63A5;&#x30D0;&#x30A4;&#x30F3;&#x30C9;&#x304A;&#x3088;&#x3073; Nck1 &#x3068; invadopodia &#x3067; cortactin &#x306E;&#x30D5;&#x30EC;&#x30C3;&#x30C8; &#x30A4;&#x30F3;&#x30BF;&#x30E9;&#x30AF;&#x30B7;&#x30E7;&#x30F3;&#x305D;&#x306E; SH2 &#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x3092;&#x4ECB;&#x3057;&#x3066;&#x52DF;&#x96C6;&#x306B;&#x3068;&#x3063;&#x3066;&#x91CD;&#x8981;&#x3067;&#x3059;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x30DE;&#x30C8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x5206;&#x89E3;&#x306B;&#x4F9D;&#x5B58;&#x3059;&#x308B;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x306F;&#x30DB;&#x30B9;&#x30DB;&#x30C1;&#x30ED;&#x30B7;&#x30F3; 421 &#x307E;&#x305F;&#x306F; 466 &#x306E;&#x7A81;&#x7136;&#x5909;&#x7570;&#x3092;&#x767A;&#x73FE;&#x3059;&#x308B;&#x7D30;&#x80DE;&#x306E;&#x5287;&#x7684;&#x306B;&#x6291;&#x5236;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x4E00;&#x7DD2;&#x306B;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x7D50;&#x679C;&#x304B;&#x3089; tyrosines 421 &#x3068; 466 cortactin &#x4E0A;&#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316; invadopodia &#x3067; Nck1 &#x4F9D;&#x5B58;&#x6027;&#x306E;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x306E;&#x91CD;&#x5408;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x91CD;&#x8981;&#x306A;&#x6B8B;&#x57FA;&#x3068;&#x3057;&#x3066;&#x8B58;&#x5225;&#x304A;&#x3088;&#x3073;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x306E;&#x6D78;&#x6F64;&#x3068;&#x5177;&#x4F53;&#x7684;&#x306B;&#x3069;&#x3061;&#x3089;&#x304B; 421 &#x307E;&#x305F;&#x306F; 466 &#x306E;&#x30C1;&#x30ED;&#x30B7;&#x30F3;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x3092;&#x30D6;&#x30ED;&#x30C3;&#x30AF; in vivo &#x816B;&#x760D;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x3092;&#x6291;&#x5236;&#x3059;&#x308B;&#x3053;&#x3068;&#x3067;&#x52B9;&#x679C;&#x7684;&#x306A;&#x3053;&#x3068;&#x304C;&#x793A;&#x5506;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;

Cortactin &#x4E0A;&#x306E;&#x7279;&#x5B9A;&#x306E;&#x30C1;&#x30ED;&#x30B7;&#x30F3;&#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x306E;&#x30B5;&#x30A4;&#x30C8; invadopodia &#x3067; Nck1 &#x4F9D;&#x5B58;&#x6027;&#x306E;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x306E;&#x91CD;&#x5408;&#x3092;&#x8ABF;&#x7BC0;&#x3057;&#x307E;&#x3059;&#x3002;

Invadopodia&#xB294; &#xD589;&#xB82C;&#xC744; &#xC800;&#xD558; &#xC2DC; &#xD0A4; &#xC9C0; &#xB9D0;&#xB77C; &#xD569;&#xC744; &#xC870;&#xC808; &#xD558;&#xB294; &#xB2E8;&#xBC31;&#xC9C8;&#xC5D0; &#xD48D;&#xBD80; &#xD558; &#xAC8C; &#xCE68; &#xC724; &#xC131; &#xC554; &#xC885; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xB9C9; &#xB3CC;&#xCD9C;. &#xC628;-&#xC624;&#xD504; &#xADDC;&#xC815; &#xC2A4;&#xC704;&#xCE58; invadopodia&#xC5D0;&#xC11C; &#xB9D0;&#xB77C; &#xC911; &#xD569;&#xC744; &#xC2DC;&#xC791; &#xD558;&#xB294; &#xD2F0;&#xB85C;&#xC2E0; &#xC794;&#xB958;&#xBB3C; 421, 466, &#xBC0F; cortactin&#xC5D0; 482&#xC758; &#xC778; &#xC0B0;&#xD654;&#xAC00; &#xD544;&#xC694; &#xD569;&#xB2C8;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;, &#xADF8;&#xAC83;&#xC740; &#xC54C; &#xC218; &#xC5C6;&#xB294;&#xC774; cortactin &#xD2F0;&#xB85C;&#xC2E0; &#xC778; &#xC0B0;&#xD654; &#xC704;&#xCE58;&#xC758; &#xB9D0;&#xB77C; &#xD569;&#xC744; &#xC81C;&#xC5B4; &#xD558;&#xB294;. &#xC6B0;&#xB9AC; cortactin invadopodia&#xC5D0;&#xC11C; &#xB9D0;&#xB77C; &#xD569; &#xADDC;&#xC815;&#xC5D0; &#xAC1C;&#xBCC4; &#xD2F0;&#xB85C;&#xC2E0; &#xC778; &#xC0B0;&#xD654; &#xC0AC;&#xC774;&#xD2B8; (421, 466, &#xBC0F; 482)&#xC758; &#xAE30;&#xC5EC;&#xB3C4; &#xC870;&#xC0AC; &#xD588;&#xB2E4;. &#xADF8; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5;&#xB294; &#xC778; &#xC0B0;&#xD654; tyrosines&#xC758; 421 466, &#xD558;&#xC9C0;&#xB9CC; &#xD558;&#xC9C0; 482&#xB294; &#xD544;&#xC694;&#xD55C; invadopodia&#xC5D0;&#xC11C; &#xBB34;&#xB8CC; &#xB9D0;&#xB77C; &#xAC00;&#xC2DC; &#xB05D;&#xC758; &#xC138;&#xB300;&#xB97C; &#xC704;&#xD55C;. &#xB610;&#xD55C;,&#xC774; &#xAC19;&#xC740; phosphotyrosines Nck1 invadopodia Nck1 cortactin &#xCCB4; &#xC678;, &#xD558;&#xC758; &#xC9C1;&#xC811; &#xBC14;&#xC778;&#xB529; &#xBC0F; nck1&#xC640; invadopodia&#xC5D0;&#xC11C; cortactin &#xC0AC;&#xC774; &#xBB34;&#xC11C; &#xC6CC; &#xC0C1;&#xD638; &#xC791;&#xC6A9;&#xC758; SH2 &#xB3C4;&#xBA54;&#xC778;&#xC744; &#xD1B5;&#xD574; &#xCC44;&#xC6A9;&#xC5D0; &#xB300; &#xD55C; &#xC911;&#xC694; &#xD558;&#xB2E4;. &#xB610;&#xD55C;, &#xB9E4;&#xD2B8;&#xB9AD;&#xC2A4; &#xBCA0;&#xC774;&#xC2A4; &#xC885;&#xC18D;&#xC801;&#xC778; &#xC885;&#xC591; &#xC138;&#xD3EC; &#xCE68;&#xACF5;&#xC740; &#xADF9;&#xC801;&#xC73C;&#xB85C; 421 &#xB610;&#xB294; 466 phosphotyrosine&#xC5D0; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xD45C;&#xD604; &#xD558;&#xB294; &#xC140;&#xC5D0; &#xC800;&#xD574; &#xB429;&#xB2C8;&#xB2E4;. &#xD568;&#xAED8;, &#xC774;&#xB7EC;&#xD55C; &#xACB0;&#xACFC; invadopodia&#xC5D0; Nck1 &#xC885;&#xC18D; &#xB9D0;&#xB77C; &#xD569;&#xC744; &#xC870;&#xC808; &#xD558;&#xB294; &#xC911;&#xC694; &#xD55C; &#xC794;&#xB958;&#xBB3C;&#xC73C;&#xB85C; 421 &#xBC0F; 466 cortactin&#xC5D0; tyrosines&#xC758; &#xC778; &#xC0B0;&#xD654;&#xB97C; &#xC2DD;&#xBCC4; &#xD558; &#xACE0; &#xC885;&#xC591; &#xC138;&#xD3EC; &#xCE68;&#xC785;&#xACFC;&#xB294; &#xD6A8;&#xACFC;&#xC801;&#xC73C;&#xB85C; &#xC885;&#xC591; &#xC138;&#xD3EC; &#xCE68;&#xACF5; vivo&#xC5D0;&#xC11C; &#xC5B5;&#xC81C; &#xB420; &#xC218; &#xC788;&#xC744; &#xD2B9;&#xBCC4;&#xD788; &#xC5B4;&#xB290; 421 &#xB610;&#xB294; 466 &#xD2F0;&#xB85C;&#xC2E0; &#xC778; &#xC0B0;&#xD654;&#xB97C; &#xCC28;&#xB2E8;.

&#xD2B9;&#xC815; &#xD2F0;&#xB85C;&#xC2E0; &#xC778; &#xC0B0;&#xD654; &#xC0AC;&#xC774;&#xD2B8; cortactin&#xC5D0; invadopodia&#xC5D0; Nck1 &#xC885;&#xC18D; &#xB9D0;&#xB77C; &#xD569; &#xC870;&#xC808;.

Invadopodios son degradantes de matriz protrusiones de membrana en las c&eacute;lulas del carcinoma invasor enriquecidas en prote&iacute;nas que regulan la polimerizaci&oacute;n de la actina. El encendido y apagado interruptor regulador que inicia la polimerizaci&oacute;n de la actina en invadopodios requiere fosforilaci&oacute;n de residuos de tirosina 421, 466 y 482 en cortactina. Sin embargo, es desconocido que de estos sitios de fosforilaci&oacute;n de tirosina cortactina controlan la polimerizaci&oacute;n de la actina. Investigamos la contribuci&oacute;n de los sitios de fosforilaci&oacute;n de tirosina individuales (421, 466 y 482) en cortactina a la regulaci&oacute;n de la polimerizaci&oacute;n de la actina en invadopodios. Proporcionamos evidencia que la fosforilaci&oacute;n de tirosinas 421 y 466, pero no 482, se requiere para la generaci&oacute;n de extremos libres actina p&uacute;as en invadopodios. Adem&aacute;s, estas mismas phosphotyrosines son importantes para el reclutamiento de Nck1 a invadopodios a trav&eacute;s de su dominio SH2, para el enlace directo de Nck1 a cortactina in vitro y para la interacci&oacute;n de traste entre Nck1 y cortactina en invadopodios. Adem&aacute;s, la invasi&oacute;n de c&eacute;lulas de tumor de prote&oacute;lisis dependiente matriz dram&aacute;ticamente es inhibida en c&eacute;lulas que expresaban una mutaci&oacute;n en phosphotyrosine 421 o 466. Juntos, estos resultados identifican fosforilaci&oacute;n de tirosinas 421 y 466 en cortactina como los residuos cruciales que regulan la polimerizaci&oacute;n de la actina Nck1-dependiente en invadopodios y tumor invasi&oacute;n de la c&eacute;lula y sugieren que puede ser eficaz en la inhibici&oacute;n in vivo de la invasi&oacute;n de la c&eacute;lula del tumor espec&iacute;ficamente bloqueando cualquier fosforilaci&oacute;n de la tirosina 421 o 466.

Sitios de fosforilaci&oacute;n de tirosina espec&iacute;fica en cortactina regulan la polimerizaci&oacute;n de la actina Nck1-dependiente en invadopodios.

&#x6D78;&#x6DA6;&#x764C;&#x7684;&#x5355;&#x5143;&#x683C;&#x4F7F;&#x7528;&#x4E13;&#x95E8;&#x7684;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x805A;&#x5408;&#x9A71;&#x52A8;&#x7A81;&#x8D77;&#x79F0;&#x4E3A; invadopodia &#x964D;&#x4F4E;&#xFF0C;&#x53EF;&#x80FD;&#x4FB5;&#x5165;&#x7EC6;&#x80DE;&#x5916;&#x57FA;&#x8D28; (ECM) &#x901A;&#x8FC7;&#x8F6C;&#x79FB;&#x3002;&#x78F7;&#x9178;&#x5316;&#x7684; invadopodium &#x86CB;&#x767D;&#x76AE;&#x5C42;&#x662F;&#x4E3B;&#x4EA4;&#x6362;&#x673A;&#xFF0C;&#x6FC0;&#x6D3B; invadopodium &#x6210;&#x719F;&#x53CA;&#x529F;&#x80FD;&#x3002;&#x76AE;&#x5C42;&#x6700;&#x521D;&#x88AB;&#x8BA4;&#x5B9A;&#x4E3A; hyperphosphorylated &#x86CB;&#x767D;&#x5728; v Src &#x8F6C;&#x6362;&#x7684;&#x5355;&#x5143;&#x683C;&#xFF0C;&#x4F46;&#x6FC0;&#x9176;&#x6216;&#x76F4;&#x63A5;&#x8D1F;&#x8D23;&#x76AE;&#x5C42;&#x78F7;&#x9178;&#x5316;&#x5728; invadopodia &#x4E2D;&#x7684;&#x6FC0;&#x9176;&#x4ECD;&#x7136;&#x672A;&#x77E5;&#x3002;&#x5728;&#x6B64;&#x7814;&#x7A76;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x63D0;&#x4F9B;&#x8BC1;&#x636E; Abl &#x76F8;&#x5173; nonreceptor &#x916A;&#x6C28;&#x9178;&#x6FC0;&#x9176; Arg &#x4ECB;&#x5BFC;&#x8868;&#x76AE;&#x751F;&#x957F;&#x56E0;&#x5B50; (EGF)-&#x8BF1;&#x5BFC;&#x76AE;&#x5C42;&#x78F7;&#x9178;&#x5316;&#xFF0C;&#x89E6;&#x53D1; invadopodia&#x3001; ECM &#x964D;&#x89E3;&#x548C;&#x77E9;&#x9635;&#x4F9D;&#x8D56;&#x86CB;&#x767D;&#x8D28;&#x964D;&#x89E3;&#x7684;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x4FB5;&#x88AD;&#x7684;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x805A;&#x5408;&#x3002;Src &#x548C; Arg &#x672C;&#x5730;&#x5316;&#x7684; invadopodia &#x548C;&#x6240;&#x9700;&#x7684;&#x8868;&#x76AE;&#x751F;&#x957F;&#x56E0;&#x5B50;&#x8BF1;&#x5BFC;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x7684;&#x805A;&#x5408;&#x3002;&#x503C;&#x5F97;&#x6CE8;&#x610F;&#x7684;&#x662F;&#xFF0C;Arg Src &#x51FB;&#x5012;&#x7EC6;&#x80DE;&#x4E2D;&#x7684;&#x9AD8;&#x6548;&#x8868;&#x8FBE;&#x90E8;&#x5206;&#x53EF;&#x4EE5;&#x62EF;&#x6551;&#x5728; invadopodia &#x4E2D;&#x7684;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x805A;&#x5408;&#x867D;&#x7136; Src &#x8868;&#x8FBE;&#x4E0D;&#x80FD;&#x8865;&#x507F;&#x635F;&#x5931;&#x7684; Arg&#xFF0C;&#x4E89;&#x8BBA; Src &#x95F4;&#x63A5;&#x8C03;&#x8282;&#x901A;&#x8FC7; Arg &#x6FC0;&#x6D3B; invadopodium &#x6210;&#x719F;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x8868;&#x660E;&#x5176;&#x4E2D; EGFR-Src-Arg-&#x76AE;&#x5C42;&#x901A;&#x8DEF;&#x4ECB;&#x5BFC;&#x7684; invadopodia &#x548C;&#x4E73;&#x817A;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x4FB5;&#x88AD;&#x7684;&#x529F;&#x80FD;&#x6210;&#x719F;&#x7684;&#x65B0;&#x673A;&#x5236;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x4ED6;&#x4EEC;&#x786E;&#x5B9A; Arg &#x4E3A; invadopodia &#x529F;&#x80FD;&#x7684;&#x65B0;&#x578B;&#x8C03;&#x89E3;&#x4EBA;&#x548C;&#x5019;&#x9009;&#x4EBA;&#x7684;&#x9776;&#x70B9;&#x6291;&#x5236;&#x80BF;&#x7624;&#x5165;&#x4FB5;&#x4F53;&#x5185;&#x3002;

&#x8868;&#x76AE;&#x751F;&#x957F;&#x56E0;&#x5B50;-Src-Arg-&#x76AE;&#x5C42;&#x901A;&#x8DEF;&#x4ECB;&#x5BFC; invadopodia &#x548C;&#x4E73;&#x817A;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x4FB5;&#x88AD;&#x7684;&#x529F;&#x80FD;&#x7684;&#x6210;&#x719F;&#x3002;

Invasive carcinoma cells use specialized actin polymerization-driven protrusions called invadopodia to degrade and possibly invade through the extracellular matrix (ECM) during metastasis. Phosphorylation of the invadopodium protein cortactin is a master switch that activates invadopodium maturation and function. Cortactin was originally identified as a hyperphosphorylated protein in v-Src-transformed cells, but the kinase or kinases that are directly responsible for cortactin phosphorylation in invadopodia remain unknown. In this study, we provide evidence that the Abl-related nonreceptor tyrosine kinase Arg mediates epidermal growth factor (EGF)-induced cortactin phosphorylation, triggering actin polymerization in invadopodia, ECM degradation, and matrix proteolysis-dependent tumor cell invasion. Both Src and Arg localize to invadopodia and are required for EGF-induced actin polymerization. Notably, Arg overexpression in Src knockdown cells can partially rescue actin polymerization in invadopodia while Src overexpression cannot compensate for loss of Arg, arguing that Src indirectly regulates invadopodium maturation through Arg activation. Our findings suggest a novel mechanism by which an EGFR-Src-Arg-cortactin pathway mediates functional maturation of invadopodia and breast cancer cell invasion. Furthermore, they identify Arg as a novel mediator of invadopodia function and a candidate therapeutic target to inhibit tumor invasion in vivo.

An EGFR-Src-Arg-cortactin pathway mediates functional maturation of invadopodia and breast cancer cell invasion.

Les cellules de carcinome invasif utilisent des protub&eacute;rances ax&eacute;e sur la polym&eacute;risation d&#39;actine sp&eacute;cialis&eacute; appel&eacute;s invadopodia pour d&eacute;grader et &eacute;ventuellement envahir par l&#39;interm&eacute;diaire de la matrice extracellulaire (ECM) au cours de la m&eacute;tastase. La phosphorylation de la cortactin de prot&eacute;ine d&#39;invadopodium est un interrupteur g&eacute;n&eacute;ral qui active la fonction et la maturation de l&#39;invadopodium. Cortactin a &eacute;t&eacute; initialement identifi&eacute;e comme &eacute;tant une prot&eacute;ine hyperphosphoryl&eacute;e dans les cellules transform&eacute;es par v-Src, mais la kinase ou kinases qui sont directement responsables de la phosphorylation de cortactin en invadopodia demeurent inconnus. Dans cette &eacute;tude, nous fournir la preuve que la tyrosine-kinase Abl li&eacute;es non r&eacute;cepteur Arg intervient dans le facteur de croissance &eacute;pidermique (EGF)-induit la phosphorylation de cortactin, d&eacute;clenchant la polym&eacute;risation de l&#39;actine dans invadopodia, d&eacute;gradation de l&#39;ECM et invasion des cellules tumorales de prot&eacute;olyse d&eacute;pendante matrice. Les Src et Arg localiser &agrave; invadopodia et sont n&eacute;cessaires pour la polym&eacute;risation de l&#39;actine induite par l&#39;EGF. Notamment, Arg surexpression dans les cellules knockdown Src peut sauver partiellement de polym&eacute;risation de l&#39;actine en invadopodia tandis que la surexpression de la Src ne peut compenser la perte de Arg, arguant que la Src r&eacute;gule indirectement invadopodium maturation gr&acirc;ce &agrave; l&#39;activation de l&#39;Arg. Nos r&eacute;sultats sugg&egrave;rent un nouveau m&eacute;canisme par lequel une voie EGFR-Src-Arg-cortactin intervient dans la maturation fonctionnelle d&#39;invasion des cellules de cancer invadopodia et cancer du sein. En outre, ils identifient Arg comme nouveau m&eacute;diateur de la fonction invadopodia et une cible th&eacute;rapeutique de candidat pour inhiber l&#39;invasion tumorale in vivo.

Une voie d&#39;EGFR-Src-Arg-cortactin intervient dans la maturation fonctionnelle d&#39;invasion des cellules de cancer invadopodia et cancer du sein.

Invasive Karzinom-Zellen verwenden spezielle Aktin Polymerisation-gesteuerte Vorspr&uuml;nge genannt Invadopodia beeintr&auml;chtigen und m&ouml;glicherweise dringen durch die extrazellul&auml;re Matrix (ECM) w&auml;hrend der Metastasierung. Phosphorylierung von der Invadopodium-Protein-Cortactin ist ein Hauptschalter, der Invadopodium-Reifung und Funktion aktiviert. Cortactin wurde urspr&uuml;nglich als Hyperphosphorylated Protein in V-Src-transformierten Zellen identifiziert, aber die Kinase oder Kinasen, die direkt verantwortlich f&uuml;r die Phosphorylierung von Cortactin in Invadopodia sind noch unbekannt. In dieser Studie wir nachweisen, dass die Tyrosinkinase Abl-bezogenen Nonreceptor Arg epidermalen Wachstumsfaktor (EGF) vermittelt-induzierte Cortactin Phosphorylierung, Aktin-Polymerisation in Invadopodia, ECM Abbau- und Matrix Proteolyse-abh&auml;ngige Tumorinvasion Zelle ausl&ouml;sen. Src und Arg zu Invadopodia zu lokalisieren und f&uuml;r EGF-induzierte Aktin-Polymerisation erforderlich sind. Insbesondere kann Arg &Uuml;berexpression in Src Niederschlag Zellen teilweise Aktin-Polymerisation in Invadopodia retten, w&auml;hrend Src &Uuml;berexpression nicht Verlust der Arg kompensieren kann, argumentierte, dass Src indirekt Invadopodium Reifung durch Arg Aktivierung reguliert. Unsere Ergebnisse weisen einen neuen Mechanismus, durch den ein EGFR-Src-Arg-Cortactin-Weg funktionelle Reifung Invadopodia und Brustkrebs Krebs Zelle Invasion vermittelt. Dar&uuml;ber hinaus identifizieren sie Arg als neuartige Vermittler der Invadopodia-Funktion und ein Kandidat Therapeutisches Zielmolek&uuml;l Tumorinvasion in-vivo zu hemmen.

Ein EGFR-Src-Arg-Cortactin-Weg vermittelt die funktionelle Reifung von Invadopodia und Brust-Krebs-Zelle-Invasion.

Cellule di carcinoma invasivo utilizzano sporgenze guidata da polimerizzazione actina specializzato chiamati invadopodia a degradare e possibilmente invadere attraverso la matrice extracellulare (ECM) durante la metastasi. La fosforilazione dei invadopodium della proteina cortactin &egrave; un interruttore che attiva la funzione e la maturazione di invadopodium. Cortactin originalmente &egrave; stato identificato come una proteina iperfosforilata nelle cellule v-Src-trasformata, ma la chinasi o chinasi che sono direttamente responsabili della fosforilazione di cortactin in invadopodia rimangono sconosciute. In questo studio, noi di fornire la prova che la tirosina chinasi Abl-correlati nonreceptor Arg da intermediario epidermal growth factor (EGF)-indotta cortactin fosforilazione, innescando la polimerizzazione dell&#39;actina in invadopodia, degradazione ECM e invasione delle cellule di matrice proteolisi dipendente dal tumore. Src sia Arg localizzare a invadopodia e sono necessari per la polimerizzazione dell&#39;actina EGF-indotta. In particolare, Arg sovraespressione in celle di colpo Src pu&ograve; salvare parzialmente polimerizzazione dell&#39;actina in invadopodia mentre Src sovraespressione non pu&ograve; compensare la perdita di Arg, sostenendo che Src regola indirettamente invadopodium maturazione attraverso l&#39;attivazione di Arg. I nostri risultati suggeriscono un romanzo meccanismo mediante il quale un pathway EGFR-Src-Arg-cortactin media maturazione funzionale dell&#39;invasione di cellule cancro seno e invadopodia. Inoltre, essi identificano Arg come un romanzo mediatore della funzione invadopodia e un obiettivo terapeutico candidato per inibire l&#39;invasione tumorale in vivo.

Un pathway EGFR-Src-Arg-cortactin media maturazione funzionale dell&#39;invasione di cellule cancro seno e invadopodia.

&#x6D78;&#x6F64;&#x764C;&#x7D30;&#x80DE; invadopodia &#x3068;&#x547C;&#x3070;&#x308C;&#x308B;&#x7279;&#x6B8A;&#x306A;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x91CD;&#x5408;&#x99C6;&#x52D5;&#x7A81;&#x8D77;&#x304C;&#x4F4E;&#x4E0B;&#x3059;&#x308B;&#x53EF;&#x80FD;&#x6027;&#x304C;&#x3042;&#x308B;&#x7D30;&#x80DE;&#x5916;&#x30DE;&#x30C8;&#x30EA;&#x30C3;&#x30AF;&#x30B9; (ECM) &#x3092;&#x901A;&#x3058;&#x3066;&#x8EE2;&#x79FB;&#x4E2D;&#x306B;&#x4FB5;&#x5165;&#x306B;&#x4F7F;&#x7528;&#x3057;&#x307E;&#x3059;&#x3002;&#x672A;&#x3060;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA; cortactin &#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x306F;&#x672A;&#x3060;&#x6210;&#x719F;&#x3068;&#x6A5F;&#x80FD;&#x3092;&#x30A2;&#x30AF;&#x30C6;&#x30A3;&#x30D6;&#x306B;&#x3059;&#x308B;&#x30DE;&#x30B9;&#x30BF;&#x30FC; &#x30B9;&#x30A4;&#x30C3;&#x30C1;&#x3067;&#x3059;&#x3002;Cortactin &#x306F;&#x3082;&#x3068;&#x3082;&#x3068;&#x904E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA; v Src &#x5909;&#x63DB;&#x7D30;&#x80DE;&#x3068;&#x3057;&#x3066;&#x8B58;&#x5225;&#x3055;&#x308C;&#x305F;&#x304C;&#x3001;&#x30AD;&#x30CA;&#x30FC;&#x30BC;&#x307E;&#x305F;&#x306F;&#x76F4;&#x63A5; cortactin &#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316; invadopodia &#x306E;&#x305F;&#x3081;&#x306B;&#x8CAC;&#x4EFB;&#x304C;&#x3042;&#x308B;&#x30AD;&#x30CA;&#x30FC;&#x30BC;&#x306F;&#x4E0D;&#x660E;&#x306E;&#x307E;&#x307E;&#x3002;&#x672C;&#x7814;&#x7A76;&#x3067;&#x306F;, Abl &#x95A2;&#x9023; nonreceptor &#x306E;&#x30C1;&#x30ED;&#x30B7;&#x30F3;&#x306E;&#x30AD;&#x30CA;&#x30FC;&#x30BC; Arg &#x4E0A;&#x76AE;&#x6210;&#x9577;&#x56E0;&#x5B50; (EGF) &#x3092;&#x4EF2;&#x4ECB;&#x3059;&#x308B;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;-&#x306E; invadopodia&#x3001;ECM &#x306E;&#x52A3;&#x5316;&#x3001;&#x30DE;&#x30C8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x5206;&#x89E3;&#x306B;&#x4F9D;&#x5B58;&#x3059;&#x308B;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x306E;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x91CD;&#x5408;&#x3092;&#x30C8;&#x30EA;&#x30AC;&#x30FC;&#x3001;cortactin &#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x3092;&#x8A98;&#x5C0E;&#x3002;Src &#x3068; Arg &#x306B; invadopodia &#x3092;&#x30ED;&#x30FC;&#x30AB;&#x30E9;&#x30A4;&#x30BA;&#x3057;&#x3001;EGF &#x8A98;&#x8D77;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x91CD;&#x5408;&#x7528;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;&#x3002;&#x7279;&#x306B;&#x3001;Arg overexpression Src &#x30CE;&#x30C3;&#x30AF;&#x30C0;&#x30A6;&#x30F3;&#x7D30;&#x80DE;&#x3067;&#x306F;&#x90E8;&#x5206;&#x7684;&#x306B; Src overexpression Src &#x306F;&#x3001;&#x76F4;&#x63A5; Arg &#x306E;&#x6D3B;&#x6027;&#x5316;&#x3092;&#x4ECB;&#x3057;&#x3066;&#x672A;&#x3060;&#x6210;&#x719F;&#x3092;&#x8ABF;&#x7BC0;&#x3057;&#x306A;&#x3044;&#x3068;&#x4E3B;&#x5F35;&#x3001;Arg &#x306E;&#x640D;&#x5931;&#x88DC;&#x511F;&#x3059;&#x308B;&#x3053;&#x3068;&#x306F;&#x3067;&#x304D;&#x307E;&#x305B;&#x3093;&#x3057;&#x306A;&#x304C;&#x3089; invadopodia &#x306E;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x91CD;&#x5408;&#x3092;&#x6551;&#x3046;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005; &#x306E;&#x8ABF;&#x67FB;&#x7D50;&#x679C;&#x3067;&#x306F; invadopodia &#x3068;&#x4E73;&#x623F;&#x306E;&#x304C;&#x3093;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x306E;&#x6A5F;&#x80FD;&#x7684;&#x6210;&#x719F; EGFR Src Arg cortactin &#x7D4C;&#x8DEF;&#x3092;&#x4EF2;&#x4ECB;&#x3059;&#x308B;&#x65B0;&#x3057;&#x3044;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3092;&#x63D0;&#x6848;&#x3057;&#x307E;&#x3059;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x5F7C;&#x3089;&#x306F; in vivo &#x3067;&#x306E;&#x816B;&#x760D;&#x6D78;&#x6F64;&#x3092;&#x6291;&#x5236;&#x3059;&#x308B; invadopodia &#x95A2;&#x6570;&#x306E;&#x5C0F;&#x8AAC;&#x30E1;&#x30C7;&#x30A3;&#x30A8;&#x30FC;&#x30BF;&#x30FC;&#x3068;&#x5019;&#x88DC;&#x8005;&#x306E;&#x6CBB;&#x7642;&#x4E0A;&#x306E;&#x30BF;&#x30FC;&#x30B2;&#x30C3;&#x30C8;&#x3068;&#x3057;&#x3066;&#x306E; Arg &#x3092;&#x8B58;&#x5225;&#x3057;&#x307E;&#x3059;&#x3002;

EGFR Src Arg cortactin &#x7D4C;&#x8DEF; invadopodia &#x3068;&#x4E73;&#x623F;&#x306E;&#x304C;&#x3093;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x306E;&#x6A5F;&#x80FD;&#x7684;&#x6210;&#x719F;&#x3092;&#x4EF2;&#x4ECB;&#x3057;&#x307E;&#x3059;&#x3002;

&#xCE68; &#xC724; &#xC131; &#xC554; &#xC885; &#xC138;&#xD3EC; invadopodia &#xB77C;&#xB294; &#xD2B9;&#xC218; &#xB9D0;&#xB77C; &#xD569; &#xAE30;&#xBC18; &#xB3CC;&#xCD9C;&#xC744; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC800;&#xD558; &#xB418; &#xACE0; &#xC804;&#xC774; &#xB3D9;&#xC548; &#xAE30;&#xC9C8; (ECM)&#xC744; &#xD1B5;&#xD574; &#xAC00;&#xB2A5; &#xD558; &#xAC8C; &#xCE68;&#xACF5;. Invadopodium &#xB2E8;&#xBC31;&#xC9C8; cortactin&#xC758; &#xC778; &#xC0B0;&#xD654;&#xB294; &#xB9C8;&#xC2A4;&#xD130; &#xC2A4;&#xC704;&#xCE58; invadopodium &#xC131;&#xC219; &#xBC0F; &#xAE30;&#xB2A5; &#xD65C;&#xC131;&#xD654;&#xC785;&#xB2C8;&#xB2E4;. Cortactin&#xAC00; &#xC6D0;&#xB798; v Src &#xBCC0;&#xD615; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; hyperphosphorylated &#xB2E8;&#xBC31;&#xC9C8;&#xC73C;&#xB85C; &#xC2DD;&#xBCC4; &#xD558;&#xC9C0;&#xB9CC; &#xD0A4; &#xB610;&#xB294; invadopodia&#xC5D0; cortactin &#xC778; &#xC0B0;&#xD654;&#xC5D0; &#xB300; &#xD55C; &#xC9C1;&#xC811; &#xCC45;&#xC784;&#xC774; kinases &#xC54C;&#xB824;&#xC9C0;&#xC9C0; &#xC54A;&#xC740; &#xB0A8;&#xC544; &#xC788;&#xB2E4;. &#xC774; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; &#xC6B0;&#xB9AC;&#xAC00; Abl &#xAD00;&#xB828; nonreceptor &#xD2F0;&#xB85C;&#xC2E0; &#xD0A4; &#xB2C8; &#xC544; &#xC81C; Arg &#xD45C; &#xD53C; &#xC131;&#xC7A5; &#xC778;&#xC790; (EGF)&#xB97C; &#xC911;&#xC7AC; &#xD558;&#xB294; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5;-invadopodia, ECM &#xC800;&#xD558;&#xC640; &#xB9E4;&#xD2B8;&#xB9AD;&#xC2A4; &#xBCA0;&#xC774;&#xC2A4; &#xC885;&#xC18D;&#xC801;&#xC778; &#xC885;&#xC591; &#xC138;&#xD3EC; &#xCE68;&#xACF5; &#xB9D0;&#xB77C; &#xD569; &#xD2B8;&#xB9AC;&#xAC70;&#xB9C1; cortactin &#xC778; &#xC0B0;&#xD654;&#xB97C; &#xC720;&#xB3C4; &#xD569;&#xB2C8;&#xB2E4;. Src&#xC640; Arg invadopodia&#xB97C; &#xC9C0;&#xC5ED;&#xD654; &#xD558; &#xACE0; EGF &#xC720;&#xB3C4; &#xB9D0;&#xB77C; &#xD569;&#xC5D0; &#xD544;&#xC694;&#xD55C;. &#xD2B9;&#xD788;, Src &#xCD5C;&#xC800;&#xC758; &#xC140;&#xC5D0; Arg overexpression Src overexpression Arg&#xB294; Src &#xC9C1;&#xC811; &#xC870;&#xC808; &#xD558;&#xC9C0; invadopodium &#xC131;&#xC219; Arg &#xD65C;&#xC131;&#xD654;&#xB97C; &#xD1B5;&#xD574; &#xB17C;&#xC7C1;&#xC758; &#xC190;&#xC2E4;&#xC5D0; &#xB300; &#xD55C; &#xBCF4;&#xC0C1; &#xC218; &#xC5C6;&#xC2B5;&#xB2C8;&#xB2E4; &#xD558;&#xB294; &#xB3D9;&#xC548; invadopodia&#xC5D0;&#xC11C; &#xB9D0;&#xB77C; &#xD569;&#xC744; &#xAD6C;&#xC870; &#xBD80;&#xBD84;&#xC801;&#xC73C;&#xB85C; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xC5F0;&#xAD6C; &#xACB0;&#xACFC;&#xB294; EGFR Src Arg cortactin &#xD1B5;&#xB85C; &#xC911;&#xC7AC; &#xAE30;&#xB2A5; &#xC131;&#xC219; invadopodia &#xBC0F; &#xC720; &#xBC29; &#xC554; &#xC138;&#xD3EC; &#xB0B4; &#xC2B5;&#xC758; &#xC0C8;&#xB85C;&#xC6B4; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC744; &#xC81C;&#xC548;. &#xB610;&#xD55C;, &#xADF8;&#xB4E4;&#xC740; &#xC885;&#xC591; &#xCE68;&#xC785; &#xC0DD;&#xCCB4; &#xC870;&#xAC74; &#xAE08;&#xC9C0; invadopodia &#xD568;&#xC218;&#xC758; &#xC18C;&#xC124; &#xC911;&#xC7AC;&#xC790; &#xD6C4;&#xBCF4; &#xCE58;&#xB8CC; &#xB300;&#xC0C1;&#xC73C;&#xB85C; Arg&#xB97C; &#xC2DD;&#xBCC4; &#xD569;&#xB2C8;&#xB2E4;.

EGFR Src Arg cortactin &#xD1B5;&#xB85C; &#xAE30;&#xB2A5; &#xC131;&#xC219; invadopodia &#xBC0F; &#xC720; &#xBC29; &#xC554; &#xC138;&#xD3EC; &#xCE68;&#xACF5;&#xC758; &#xC911;&#xC7AC;.

Las c&eacute;lulas del carcinoma invasor utilizan salientes de basadas en la polimerizaci&oacute;n de actina especializado llamados invadopodios para degradar y posiblemente invadir a trav&eacute;s de la matriz extracelular (MEC) durante la met&aacute;stasis. Fosforilaci&oacute;n de la cortactina de la prote&iacute;na de invadopodium es un interruptor que activa la funci&oacute;n y maduraci&oacute;n invadopodium. Cortactina fue identificado originalmente como una prote&iacute;na hiperfosforilada en c&eacute;lulas v-Src-transformado, pero desconocemos la cinasa o quinasas que son directamente responsables de la fosforilaci&oacute;n de cortactina en invadopodios. En este estudio, Proporcionamos evidencia que la tirosincinasa nonreceptor relacionados con Abl Arg media factor de crecimiento epid&eacute;rmico (EGF)-inducida por fosforilaci&oacute;n cortactina, desencadenando la polimerizaci&oacute;n de la actina en invadopodios, la degradaci&oacute;n de la ECM y la invasi&oacute;n de c&eacute;lulas de tumor de prote&oacute;lisis dependiente matriz. Fuente tanto Arg localizaci&oacute;n a invadopodios y se requieren para la polimerizaci&oacute;n de la actina EGF-inducida. En particular, sobreexpresi&oacute;n de Arg en c&eacute;lulas precipitaci&oacute;n fuente parcialmente puede rescatar la polimerizaci&oacute;n de la actina en invadopodios mientras que la sobreexpresi&oacute;n de la fuente no puede compensar p&eacute;rdida de Arg, argumentando que fuente indirectamente regula la maduraci&oacute;n de la invadopodium a trav&eacute;s de la activaci&oacute;n de Arg. Nuestros resultados sugieren un nuevo mecanismo por el cual un camino de EGFR-fuente-Arg-cortactina media maduraci&oacute;n funcional de la invasi&oacute;n de c&eacute;lulas de c&aacute;ncer invadopodios y mama. Adem&aacute;s, se identifica Arg como un mediador de la novela de invadopodios funci&oacute;n y una diana terap&eacute;utica del candidato para inhibir la invasi&oacute;n tumoral in vivo.

Una v&iacute;a de EGFR-fuente-Arg-cortactina media maduraci&oacute;n funcional de la invasi&oacute;n de c&eacute;lulas de c&aacute;ncer invadopodios y mama.

RhoGTPases haben bei der Regulierung der Krebsmetastase verwickelt. Invasive Karzinom-Zellen bilden Invadopodia, F-Aktin-reiche Matrix-erniedrigende Vorspr&uuml;nge, die man f&uuml;r Tumor wichtig sein Handy-Invasion und Intravasation. Regulierung des Aktin-Dynamik bei Invadopodial Vorspr&uuml;nge ist mit dem Einmarsch entscheidend. Dieser Prozess erfordert die ausgel&ouml;st Aktivit&auml;t von Cofilin Aktin-freie Stacheldraht enden zu generieren. Bisherige Arbeit zeigt, dass der Cofilin ausgel&ouml;st Aktivit&auml;t eng durch mehrere Mechanismen, einschlie&szlig;lich der Aufsicht &uuml;ber Cofilin Serin Phosphorylierung von Rho-GTPasen geregelt wird. Jedoch ist es nicht bekannt, die Rho GTPase ist beteiligt an der Regulierung von Cofilin Phosphorylierung Status bei Invadopodia.

Eine neuartige raumzeitliche RhoC Weg Aktivierung lokal reguliert Cofilin T&auml;tigkeit am Invadopodia.

RhoGTPases sono stati implicati nella regolazione delle metastasi del cancro. Cellule di carcinoma invasivo forma invadopodia, ricchi di F-actina protrusioni matrix-degradante che si pensiero siano importanti per il tumore delle cellule di invasione e intravasation. Regolamento delle dinamiche di actina a protrusioni invadopodial &egrave; cruciale per invasione di auto. Questo processo richiede la rottura attivit&agrave; di cofilin per generare estremit&agrave; spinata privo di actina. Lavoro precedente dimostra che attivit&agrave; di rottura di cofilin &egrave; strettamente regolamentata attraverso meccanismi multipli, tra cui regolazione della fosforilazione della serina cofilin da Rho GTPases. Tuttavia, non &egrave; noto quale GTPasi Rho &egrave; coinvolto nella regolazione dello stato di fosforilazione di cofilin a invadopodia.

Un romanzo spatiotemporal RhoC attivazione percorso localmente regola l&#39;attivit&agrave; di cofilin a invadopodia.

RhoGTPases &#x6709;&#x7275;&#x8FDE;&#x7684;&#x764C;&#x75C7;&#x8F6C;&#x79FB;&#x89C4;&#x7BA1;&#x3002;&#x4FB5;&#x88AD;&#x6027;&#x764C;&#x7EC6;&#x80DE;&#x5F62;&#x6210; invadopodia&#xFF0C;F-&#x808C;&#x52A8;&#x86CB;&#x767D;&#x5BCC;&#x77E9;&#x9635;&#x6709;&#x8FB1;&#x4EBA;&#x683C;&#x7684;&#x7A81;&#x8D77;&#xFF0C;&#x88AB;&#x8BA4;&#x4E3A;&#x662F;&#x91CD;&#x8981;&#x7684;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x5165;&#x4FB5;&#x548C;&#x5165;&#x4FB5;&#x3002;&#x5728; invadopodial &#x7A81;&#x8D77;&#x7684;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x52A8;&#x529B;&#x5B66;&#x7684;&#x8C03;&#x8282;&#x662F;&#x5173;&#x952E;&#x9A71;&#x52A8;&#x5668;&#x5165;&#x4FB5;&#x3002;&#x8FD9;&#x4E00;&#x8FC7;&#x7A0B;&#x8981;&#x6C42;&#x5207;&#x65AD; cofilin &#x751F;&#x6210;&#x65E0;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x7684;&#x5012;&#x523A;&#x7684;&#x7ED3;&#x675F;&#x7684;&#x6D3B;&#x52A8;&#x3002;&#x4EE5;&#x524D;&#x7684;&#x5DE5;&#x4F5C;&#x6F14;&#x793A; cofilin &#x7684;&#x5272;&#x65AD;&#x6D3B;&#x52A8;&#x4E25;&#x5BC6;&#x7684;&#x76D1;&#x7BA1;&#xFF0C;&#x901A;&#x8FC7;&#x591A;&#x4E2A;&#x673A;&#x5236;&#xFF0C;&#x5305;&#x62EC; cofilin &#x4E1D;&#x6C28;&#x9178;&#x78F7;&#x9178;&#x5316;&#x9176;&#x7684;&#x8C03;&#x8282;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x5B83;&#x4E0D;&#x77E5;&#x9053;&#x8FD9; Rho &#x86CB;&#x767D;&#x662F;&#x8D1F;&#x8D23;&#x89C4;&#x7BA1;&#x5728; invadopodia cofilin &#x7684;&#x78F7;&#x9178;&#x5316;&#x72B6;&#x6001;&#x3002;

&#x5C0F;&#x8BF4;&#x65F6;&#x7A7A; RhoC &#x6FC0;&#x6D3B;&#x901A;&#x8DEF;&#x672C;&#x5730;&#x89C4;&#x7BA1; invadopodia cofilin &#x6D3B;&#x52A8;&#x3002;

RhoGTPases have been implicated in the regulation of cancer metastasis. Invasive carcinoma cells form invadopodia, F-actin-rich matrix-degrading protrusions that are thought to be important for tumor cell invasion and intravasation. Regulation of actin dynamics at invadopodial protrusions is crucial to drive invasion. This process requires the severing activity of cofilin to generate actin-free barbed ends. Previous work demonstrates that cofilin's severing activity is tightly regulated through multiple mechanisms, including regulation of cofilin serine phosphorylation by Rho GTPases. However, it is not known which Rho GTPase is involved in regulating cofilin's phosphorylation status at invadopodia.

A novel spatiotemporal RhoC activation pathway locally regulates cofilin activity at invadopodia.

RhoGTPases ont &eacute;t&eacute; impliqu&eacute;s dans la r&eacute;gulation des m&eacute;tastases canc&eacute;reuses. Les cellules de carcinome invasif forment invadopodia, riches en actine-F d&eacute;gradant la matrice des protub&eacute;rances qui sont consid&eacute;r&eacute;s comme importants pour tumeur &agrave; cellules invasion et intravasation. R&egrave;glement des dynamiques de l&#39;actine au invadopodial saillies est cruciale pour l&#39;invasion en voiture. Ce processus requiert l&#39;activit&eacute; de sectionnement de cofilin pour g&eacute;n&eacute;rer les extr&eacute;mit&eacute;s barbel&eacute;es exempte d&#39;actine. Des travaux ant&eacute;rieurs montre que l&#39;activit&eacute; sectionnement de cofilin est &eacute;troitement contr&ocirc;l&eacute;e par le biais de plusieurs m&eacute;canismes, y compris la r&eacute;glementation de la phosphorylation de s&eacute;rine cofilin par GTPases Rho. Cependant, on ne sait pas qui est de la Rho GTPase impliqu&eacute;s dans la r&eacute;gulation des &Eacute;tat de phosphorylation de cofilin &agrave; invadopodia.

Une roman spatio-temporelle RhoC voie d&#39;activation localement r&eacute;glemente l&#39;activit&eacute; de cofilin &agrave; invadopodia.

RhoGTPases &#x306F;&#x3001;&#x764C;&#x306E;&#x8EE2;&#x79FB;&#x306E;&#x8ABF;&#x7BC0;&#x306B;&#x95A2;&#x4E0E;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x6D78;&#x6F64;&#x764C;&#x7D30;&#x80DE;&#x3092;&#x5F62;&#x6210;&#x3059;&#x308B; invadopodia&#x3001;F &#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x304C;&#x8C4A;&#x5BCC;&#x306A;&#x816B;&#x760D;&#x306B;&#x5BFE;&#x3057;&#x3066;&#x91CD;&#x8981;&#x3068;&#x8003;&#x3048;&#x3089;&#x308C;&#x308B;&#x30DE;&#x30C8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x5206;&#x89E3;&#x7A81;&#x8D77;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x3068; intravasation&#x3002;&#x8ABF;&#x7BC0;&#x30A2;&#x30AF;&#x30C1;&#x30F3; &#x30FB; &#x30C0;&#x30A4;&#x30CA;&#x30DF;&#x30AF;&#x30B9; invadopodial &#x7A81;&#x8D77;&#x3067;&#x306E;&#x30C9;&#x30E9;&#x30A4;&#x30D6;&#x306E;&#x4FB5;&#x7565;&#x306B;&#x91CD;&#x5927;&#x3067;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306F;&#x65AD;&#x7D76;&#x3057;&#x3088;&#x3046;&#x6D3B;&#x52D5;&#x30B3;&#x30D5;&#x30A3;&#x30EA;&#x30F3;&#x306F;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x7121;&#x6599;&#x30D0;&#x30FC;&#x30D6;&#x306E;&#x4E21;&#x7AEF;&#x3092;&#x751F;&#x6210;&#x3059;&#x308B;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;&#x3002;&#x524D;&#x306E;&#x4ED5;&#x4E8B;&#x306B;&#x304A;&#x3051;&#x308B; Rho gtp &#x30A2;&#x30FC;&#x30BC;&#x306B;&#x3088;&#x308B;&#x30B3;&#x30D5;&#x30A3;&#x30EA;&#x30F3;&#x30EA;&#x30F3; &#x30BB;&#x30EA;&#x30F3;&#x306E;&#x898F;&#x5236;&#x306A;&#x3069;&#x3001;&#x8907;&#x6570;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3092;&#x901A;&#x3057;&#x3066;&#x30B3;&#x30D5;&#x30A3;&#x30EA;&#x30F3;&#x306E;&#x65AD;&#x7D76;&#x3057;&#x3088;&#x3046;&#x6D3B;&#x52D5;&#x304C;&#x5805;&#x304F;&#x8ABF;&#x6574;&#x3055;&#x308C;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x3057;&#x304B;&#x3057;&#x3001;&#x305D;&#x308C;&#x306F; Rho GTPase &#x3092;&#x77E5;&#x3089;&#x308C;&#x3066;&#x3044;&#x306A;&#x3044;&#x30B3;&#x30D5;&#x30A3;&#x30EA;&#x30F3;&#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x72B6;&#x614B;&#x3067; invadopodia &#x306E;&#x8ABF;&#x7BC0;&#x306B;&#x95A2;&#x4E0E;&#x3057;&#x307E;&#x3059;&#x3002;

&#x6642;&#x7A7A;&#x9593; RhoC &#x6D3B;&#x6027;&#x5316;&#x7D4C;&#x8DEF;&#x30ED;&#x30FC;&#x30AB;&#x30EB; invadopodia &#x30B3;&#x30D5;&#x30A3;&#x30EA;&#x30F3;&#x6D3B;&#x52D5;&#x3092;&#x8ABF;&#x7BC0;&#x3057;&#x307E;&#x3059;&#x3002;

RhoGTPases &#xC554; &#xC804;&#xC774;&#xC758; &#xB808; &#xADE4; &#xB808;&#xC774; &#xC158;&#xC5D0; &#xC5F0;&#xB8E8; &#xB418;&#xC5B4; &#xC788;&#xB2E4;. &#xCE68; &#xC724; &#xC131; &#xC554; &#xC885; &#xC138;&#xD3EC; &#xD615;&#xC131; &#xD55C;&#xB2E4; invadopodia, F &#xB9D0;&#xB77C; &#xD48D;&#xBD80;&#xD55C; &#xC885;&#xC591;&#xC5D0; &#xB300; &#xD55C; &#xC911;&#xC694; &#xD55C; &#xAC83;&#xC73C;&#xB85C; &#xC0DD;&#xAC01; &#xB418;&#xB294; &#xB9E4;&#xD2B8;&#xB9AD;&#xC2A4; &#xC800;&#xD558; &#xB3CC;&#xCD9C; &#xC138;&#xD3EC; &#xCE68;&#xACF5; &#xBC0F; intravasation. Invadopodial &#xB3CC;&#xCD9C;&#xC5D0;&#xC11C; &#xB9D0;&#xB77C; &#xC5ED;&#xD559;&#xC758; &#xB808; &#xADE4; &#xB808;&#xC774; &#xC158; &#xB4DC;&#xB77C;&#xC774;&#xBE0C; &#xCE68;&#xB7B5;&#xC5D0; &#xACB0;&#xC815;&#xC801; &#xC774;&#xB2E4;. &#xC774; &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xC5D0;&#xC11C;&#xB294; &#xB9D0;&#xB77C; &#xD504;&#xB9AC; &#xAC00;&#xC2DC; &#xB05D;&#xC744; &#xC0DD;&#xC131; &#xD558;&#xB294; cofilin&#xC758; severing &#xD65C;&#xB3D9;. &#xC774;&#xC804; &#xC791;&#xC5C5; cofilin&#xC758; severing &#xD65C;&#xB3D9; cofilin &#xB5A0;&#xB4E4;&#xACE0; &#xC778; &#xC0B0;&#xD654;&#xB85C; Gtpases&#xC5D0; &#xC758;&#xD574; &#xADDC;&#xC81C;&#xB97C; &#xD3EC;&#xD568; &#xD558; &#xC5EC; &#xC5EC;&#xB7EC; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC744; &#xD1B5;&#xD574; &#xC5C4;&#xACA9; &#xD558; &#xAC8C; &#xADDC;&#xC81C; &#xD558;&#xB294; &#xBC29;&#xBC95;&#xC744; &#xBCF4;&#xC5EC; &#xC90D;&#xB2C8;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;, &#xBAA8;&#xB978;&#xB2E4;&#xB85C; Gtpase&#xC740; invadopodia&#xC5D0;&#xC11C; cofilin&#xC758; &#xC778; &#xC0B0;&#xD654; &#xC0C1;&#xD0DC; &#xC870;&#xC808;&#xC5D0; &#xAD00;&#xC5EC; &#xD569;&#xB2C8;&#xB2E4;.

&#xC18C;&#xC124; spatiotemporal RhoC &#xD65C;&#xC131;&#xD654; &#xD1B5;&#xB85C; &#xB85C;&#xCEEC; invadopodia&#xC5D0; cofilin &#xD65C;&#xB3D9;&#xC744; &#xC870;&#xC808;.

RhoGTPases han sido implicados en la regulaci&oacute;n de la met&aacute;stasis del c&aacute;ncer. Forma de las c&eacute;lulas de carcinoma invasor invadopodios, F-actina-rico protuberancias degradadoras de matriz que se cree que es importante para el tumor de c&eacute;lulas invasi&oacute;n e intravasaci&oacute;n. Regulaci&oacute;n de la din&aacute;mica de la actina en invadopodial salientes es crucial a la invasi&oacute;n de la unidad. Este proceso requiere la actividad seccionamiento de cofilin para generar extremos p&uacute;as libre de actina. Trabajo previo demuestra que actividad seccionamiento de cofilin es estrictamente regulado a trav&eacute;s de m&uacute;ltiples mecanismos, incluyendo la regulaci&oacute;n de la fosforilaci&oacute;n de serina cofilin por GTPasas Rho. Sin embargo, no se sabe que es Rho GTPasa implicada en la regulaci&oacute;n del estado de fosforilaci&oacute;n de cofilin en invadopodios.

Una novela spatiotemporal RhoC activaci&oacute;n v&iacute;a localmente regula la actividad de cofilin en invadopodios.

Invadopodia &#x662F;&#x552F;&#x4E00;&#x5728;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x4E2D;&#x53D1;&#x73B0;&#x7684;&#x86CB;&#x767D;&#x6C34;&#x89E3;&#x6D3B;&#x52A8;&#x4E0E;&#x521B;&#x7A81;&#x8D77;&#x3002;&#x76AE;&#x5C42;&#x78F7;&#x9178;&#x5316; invadopodia &#x6210;&#x719F;&#xFF0C;&#x8C03;&#x8282; Nck1 &#x7ED1;&#x5B9A;&#x548C; cofilin &#x6D3B;&#x52A8;&#x671F;&#x95F4;&#x7684;&#x4E00;&#x4E2A;&#x5173;&#x952E;&#x6B65;&#x9AA4;&#x3002;&#x4E0D;&#x5145;&#x5206;&#x7406;&#x89E3;&#x76AE;&#x5C42;&#x4F9D;&#x8D56; cofilin &#x8C03;&#x8282;&#x7684;&#x786E;&#x5207;&#x673A;&#x7406;&#x548C;&#x6B64;&#x8DEF;&#x5F84;&#x5728; invadopodia &#x6210;&#x719F;&#x548C;&#x7EC6;&#x80DE;&#x5165;&#x4FB5;&#x4E2D;&#x7684;&#x89D2;&#x8272;&#x3002;&#x6211;&#x4EEC;&#x63D0;&#x4F9B;&#x8BC1;&#x636E;&#xFF0C;&#x76AE;&#x5C42; cofilin &#x7ED1;&#x5B9A;&#x53D7;&#x5728; invadopodia &#x4ECB;&#x5BFC; NHE1 &#x94A0;&#x6C22;&#x6362;&#x70ED;&#x5668;&#x7684;&#x672C;&#x5730; ph &#x503C;&#x53D8;&#x5316;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x76AE;&#x5C42;&#x916A;&#x6C28;&#x9178;&#x78F7;&#x9178;&#x5316;&#x4ECB;&#x5BFC; invadopodium &#x8F66;&#x53A2;&#xFF0C;&#x672C;&#x5730;&#x589E;&#x52A0;&#x4E86; ph &#x503C;&#xFF0C;&#x5BFC;&#x81F4;&#x91CA;&#x653E;&#x7684; cofilin &#x4ECE;&#x76AE;&#x5C42;&#x7684; NHE1 &#x62DB;&#x52DF;&#x4EBA;&#x5458;&#x3002;&#x6211;&#x4EEC;&#x663E;&#x793A;&#x6D89;&#x53CA;&#x76AE;&#x5C42;&#x78F7;&#x9178;&#x5316;&#x3001; &#x5C40;&#x90E8; ph &#x503C;&#x589E;&#x52A0;&#x548C; cofilin &#x6FC0;&#x6D3B;&#x8FD9;&#x4E00;&#x673A;&#x5236;&#x8C03;&#x8282; invadopodium &#x690E;&#x95F4;&#x76D8;&#x7A81;&#x51FA;&#x548C;&#x9000;&#x7F29;&#x7684;&#x52A8;&#x6001;&#x5468;&#x671F;&#x548C;&#x5BF9;&#x4E8E; 3d &#x7EC6;&#x80DE;&#x5165;&#x4FB5;&#x81F3;&#x5173;&#x91CD;&#x8981;&#x3002;&#x5728;&#x4E00;&#x8D77;&#xFF0C;&#x8FD9;&#x4E9B;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x786E;&#x5B9A;&#x65B0;&#x578B; pH &#x4F9D;&#x8D56;&#x578B;&#x8C03;&#x63A7;&#x7684;&#x7EC6;&#x80DE;&#x5165;&#x4FB5;&#x3002;

&#x76AE;&#x5C42;&#x78F7;&#x9178;&#x5316;&#x8C03;&#x8282;&#x7EC6;&#x80DE;&#x5165;&#x4FB5; pH &#x4F9D;&#x8D56;&#x9014;&#x5F84;&#x3002;

Invadopodia are invasive protrusions with proteolytic activity uniquely found in tumor cells. Cortactin phosphorylation is a key step during invadopodia maturation, regulating Nck1 binding and cofilin activity. The precise mechanism of cortactin-dependent cofilin regulation and the roles of this pathway in invadopodia maturation and cell invasion are not fully understood. We provide evidence that cortactin-cofilin binding is regulated by local pH changes at invadopodia that are mediated by the sodium-hydrogen exchanger NHE1. Furthermore, cortactin tyrosine phosphorylation mediates the recruitment of NHE1 to the invadopodium compartment, where it locally increases the pH to cause the release of cofilin from cortactin. We show that this mechanism involving cortactin phosphorylation, local pH increase, and cofilin activation regulates the dynamic cycles of invadopodium protrusion and retraction and is essential for cell invasion in 3D. Together, these findings identify a novel pH-dependent regulation of cell invasion.

Cortactin phosphorylation regulates cell invasion through a pH-dependent pathway.

Invadopodia sont envahissantes saillies avec activit&eacute; prot&eacute;olytique unique trouv&eacute; dans les cellules tumorales. Cortactin la phosphorylation est une &eacute;tape cl&eacute; au cours de la maturation invadopodia, r&eacute;gulation de liaison Nck1 et activit&eacute; cofilin. Le m&eacute;canisme pr&eacute;cis de r&egrave;glement cofilin cortactin-d&eacute;pendante et les r&ocirc;les de cette voie en invadopodia invasion de maturation et les cellules ne sont pas totalement comprises. Nous d&eacute;montrons que la liaison cortactin-cofilin est r&eacute;gul&eacute;e par les changements de pH local &agrave; invadopodia qui sont v&eacute;hicul&eacute;es par l&#39;&eacute;changeur de sodium-hydrog&egrave;ne NHE1. En outre, la phosphorylation de tyrosine cortactin intervient dans le recrutement de NHE1 dans le compartiment d&#39;invadopodium, o&ugrave; il augmente localement le pH &agrave; provoquer la lib&eacute;ration de cofilin de cortactin. Nous montrons que ce m&eacute;canisme impliquant la phosphorylation de cortactin, augmentation du pH local et cofilin activation r&eacute;gule les cycles dynamiques de protrusion invadopodium et r&eacute;traction et qu&#39;il est essentiel pour l&#39;invasion de la cellule en 3D. Ces r&eacute;sultats identifient un nouveau r&egrave;glement d&eacute;pendante du pH d&#39;invasion des cellules.

Cortactin phosphorylation r&eacute;gule invasion des cellules par une voie d&eacute;pendante du pH.

Invadopodia sind invasive Vorspr&uuml;nge mit proteolytische Aktivit&auml;t eindeutig in Tumorzellen gefunden. Cortactin Phosphorylierung ist ein wichtiger Schritt, w&auml;hrend Invadopodia Reifung, Regulierung, Nck1 Bindung und Cofilin Aktivit&auml;t. Der genaue Mechanismus der Cortactin-abh&auml;ngige Cofilin-Verordnung und die Rollen von diesem Weg Invadopodia Reifung und Zelle Invasion sind nicht vollst&auml;ndig verstanden. Wir belegen, dass Cortactin-Cofilin Bindung von den lokalen pH-Wert-Ver&auml;nderungen im Invadopodia geregelt ist, die durch die Natrium-Wasserstoff-W&auml;rmetauscher NHE1 vermittelt werden. Dar&uuml;ber hinaus vermittelt die Cortactin Tyrosin-Phosphorylierung die Rekrutierung von NHE1 ins Invadopodium, wo es lokal den pH-Wert erh&ouml;ht, die Freisetzung von Cofilin aus Cortactin zu verursachen. Wir zeigen, dass dieser Mechanismus Cortactin Phosphorylierung, lokalen pH-Wert-Anstieg und Cofilin Aktivierung die dynamischen Zyklen der Invadopodium K&ouml;rper und Retraktion regelt und essentiell f&uuml;r Zelle Invasion in 3D. Gemeinsam kennzeichnen diese Erkenntnisse eine neuartige pH-Wert-abh&auml;ngige Verordnung Zelle Invasion.

Cortactin Phosphorylierung reguliert Zelle Invasion durch eine pH-Wert-abh&auml;ngige-Pathway.

Invadopodia sono invasivi protrusioni con attivit&agrave; proteolitica in modo univoco trovato nelle cellule tumorali. Cortactin fosforilazione &egrave; un passo fondamentale durante la maturazione invadopodia, associazione di Nck1 e cofilin attivit&agrave; di regolazione. Il meccanismo preciso di regolamento cofilin cortactin-dipendente e i ruoli di questa via metabolica in invadopodia invasione di maturazione e la cella non sono completamente chiari. Forniamo prove che cortactin-cofilin associazione &egrave; regolata da cambiamenti di pH locale a invadopodia che sono mediati dallo scambiatore NHE1 di sodio-idrogeno. Inoltre, la fosforilazione della tirosina cortactin media l&#39;assunzione di NHE1 per lo scomparto di invadopodium, dove localmente aumenta il pH a causare il rilascio di cofilin da cortactin. Mostriamo che questo meccanismo che coinvolge la fosforilazione cortactin, aumento del pH locale e attivazione cofilin regola i cicli dinamici di invadopodium protrusione e retrazione ed &egrave; essenziale per l&#39;invasione delle cellule in 3D. Insieme, questi risultati identificano un romanzo regolamento pH-dipendente dell&#39;invasione delle cellule.

Fosforilazione Cortactin regola invasione delle cellule attraverso un percorso di pH-dipendente.

Invadopodia &#x4FB5;&#x8972;&#x306E;&#x7A81;&#x8D77;&#x306E;&#x30D7;&#x30ED;&#x30C6;&#x30A2;&#x30FC;&#x30BC;&#x6D3B;&#x6027;&#x3092;&#x4E00;&#x610F;&#x306B;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x306E;&#x767A;&#x898B;&#x3067;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;Cortactin &#x30EA;&#x30F3;&#x9178;&#x5316;&#x306E;&#x91CD;&#x8981;&#x306A;&#x30B9;&#x30C6;&#x30C3;&#x30D7;&#x306F;&#x3001;Nck1 &#x30D0;&#x30A4;&#x30F3;&#x30C7;&#x30A3;&#x30F3;&#x30B0;&#x304A;&#x3088;&#x3073;&#x30B3;&#x30D5;&#x30A3;&#x30EA;&#x30F3;&#x6D3B;&#x52D5;&#x3092;&#x898F;&#x5236;&#x3059;&#x308B; invadopodia &#x6210;&#x719F;&#x904E;&#x7A0B;&#x3067;&#x3059;&#x3002;Cortactin &#x4F9D;&#x5B58;&#x30B3;&#x30D5;&#x30A3;&#x30EA;&#x30F3;&#x898F;&#x5236;&#x306E;&#x6B63;&#x78BA;&#x306A;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3068;&#x3053;&#x306E; invadopodia &#x6210;&#x719F;&#x3068;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x306B;&#x304A;&#x3051;&#x308B;&#x7D4C;&#x8DEF;&#x306E;&#x5F79;&#x5272;&#x306F;&#x5B8C;&#x5168;&#x306B;&#x308F;&#x304B;&#x308A;&#x307E;&#x305B;&#x3093;&#x3002;Cortactin &#x30B3;&#x30D5;&#x30A3;&#x30EA;&#x30F3;&#x7D50;&#x5408;&#x6C34;&#x7D20;&#x30CA;&#x30C8;&#x30EA;&#x30A6;&#x30E0; &#x30A8;&#x30AF;&#x30B9;&#x30C1;&#x30A7;&#x30F3; &#x30B8;&#x30E3;&#x30FC; NHE1 &#x306B;&#x3088;&#x3063;&#x3066;&#x4EF2;&#x4ECB;&#x3055;&#x308C;&#x308B;&#x30ED;&#x30FC;&#x30AB;&#x30EB; pH &#x5909;&#x5316;&#x3067; invadopodia &#x306B;&#x3088;&#x3063;&#x3066;&#x8ABF;&#x7BC0;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x3053;&#x3068;&#x306E;&#x8A3C;&#x62E0;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;cortactin &#x30C1;&#x30ED;&#x30B7;&#x30F3;&#x30EA;&#x30F3;&#x9178;&#x5316; NHE1 &#x306E;&#x52DF;&#x96C6;&#x305D;&#x308C;&#x30ED;&#x30FC;&#x30AB;&#x30EB; cortactin &#x304B;&#x3089;&#x30B3;&#x30D5;&#x30A3;&#x30EA;&#x30F3;&#x306E;&#x30EA;&#x30EA;&#x30FC;&#x30B9;&#x3092;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3059;&#x306B; pH &#x3092;&#x5897;&#x52A0;&#x3001;&#x672A;&#x3060;&#x30B3;&#x30F3;&#x30D1;&#x30FC;&#x30C8;&#x30E1;&#x30F3;&#x30C8;&#x3092;&#x4EF2;&#x4ECB;&#x3057;&#x307E;&#x3059;&#x3002;Cortactin &#x30EA;&#x30F3;&#x9178;&#x5316;&#x3001;&#x30ED;&#x30FC;&#x30AB;&#x30EB;&#x306E; pH &#x5897;&#x52A0;&#x30B3;&#x30D5;&#x30A3;&#x30EA;&#x30F3; &#x30A2;&#x30AF;&#x30C6;&#x30A3;&#x30D9;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x3092;&#x542B;&#x3080;&#x3053;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x306F;&#x672A;&#x3060;&#x7A81;&#x8D77;&#x3068;&#x64A4;&#x56DE;&#x306E;&#x52D5;&#x7684;&#x306A;&#x30B5;&#x30A4;&#x30AF;&#x30EB;&#x3092;&#x8ABF;&#x7BC0;&#x3057;&#x3066;&#x7D30;&#x80DE;&#x4FB5;&#x5165; 3 D &#x306E;&#x305F;&#x3081;&#x306B;&#x4E0D;&#x53EF;&#x6B20;&#x3067;&#x3059;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x4E00;&#x7DD2;&#x306B;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x77E5;&#x898B;&#x306F;&#x65B0;&#x898F;&#x306E; pH &#x4F9D;&#x5B58;&#x6027;&#x8ABF;&#x7BC0;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x3092;&#x8B58;&#x5225;&#x3057;&#x307E;&#x3059;&#x3002;

Cortactin &#x30EA;&#x30F3;&#x9178;&#x5316; pH &#x4F9D;&#x5B58;&#x6027;&#x7D4C;&#x8DEF;&#x3092;&#x4ECB;&#x3057;&#x3066;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x3092;&#x8ABF;&#x7BC0;&#x3057;&#x307E;&#x3059;&#x3002;

Invadopodia&#xB294; &#xB2E8;&#xBC31;&#xC9C8; &#xD65C;&#xB3D9; &#xACE0;&#xC720; &#xD558; &#xAC8C; &#xC885;&#xC591; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xBC1C;&#xACAC;&#xC73C;&#xB85C; &#xCE68;&#xB7B5; &#xC801; &#xB3CC;&#xCD9C;. Cortactin &#xC778; &#xC0B0;&#xD654; invadopodia &#xC131;&#xC219;, Nck1 &#xBC14;&#xC778;&#xB529; &#xBC0F; cofilin &#xD65C;&#xB3D9;&#xC744; &#xADDC;&#xC81C; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xC911;&#xC694; &#xD55C; &#xB2E8;&#xACC4;&#xC785;&#xB2C8;&#xB2E4;. Cortactin &#xC885;&#xC18D; cofilin &#xADDC;&#xC815;&#xC758; &#xC815;&#xD655;&#xD55C; &#xBA54;&#xCEE4;&#xB2C8;&#xC998; &#xBC0F; invadopodia &#xC131;&#xC219;&#xACFC; &#xC140; &#xCE68;&#xACF5;&#xC5D0;&#xC774; &#xD1B5;&#xB85C;&#xC758; &#xC5ED;&#xD560;&#xC740; &#xC644;&#xC804;&#xD788; &#xC774;&#xD574; &#xB418;&#xC9C0; &#xC54A;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC; cortactin cofilin &#xBC14;&#xC778;&#xB529;&#xC774; &#xB098;&#xD2B8;&#xB968; &#xC218;&#xC18C; &#xAD50;&#xD658;&#xAE30; nhe1&#xC5D0; &#xC758;&#xD574; &#xC911;&#xC7AC; invadopodia&#xC5D0; &#xB85C;&#xCEEC; pH &#xBCC0;&#xD654;&#xC5D0; &#xC758;&#xD574; &#xADDC;&#xC81C; &#xD558;&#xB294; &#xC99D;&#xAC70;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, cortactin &#xD2F0;&#xB85C;&#xC2E0; &#xC778; &#xC0B0;&#xD654; &#xC911;&#xC7AC; &#xC5B4;&#xB514; &#xB85C;&#xCEEC; cortactin&#xC5D0;&#xC11C; cofilin&#xC758; &#xCD9C;&#xC2DC;&#xB97C; pH &#xC99D;&#xAC00; invadopodium &#xAD6C;&#xD68D;&#xC744; nhe1&#xC758; &#xCC44;&#xC6A9;. &#xC6B0;&#xB9AC; cortactin &#xC778; &#xC0B0;&#xD654;, &#xB85C;&#xCEEC; pH &#xC99D;&#xAC00;, &#xADF8;&#xB9AC;&#xACE0; cofilin &#xD65C;&#xC131;&#xD654;&#xAC00;&#xC774; &#xBA54;&#xCEE4;&#xB2C8;&#xC998; invadopodium &#xB3CC;&#xCD9C; &#xBC0F; &#xCCA0;&#xD68C;&#xC758; &#xB3D9;&#xC801; &#xC0AC;&#xC774;&#xD074;&#xC744; &#xC870;&#xC808; &#xD558; &#xACE0; 3 &#xCC28;&#xC6D0;&#xC5D0;&#xC11C; &#xC140; &#xCE68;&#xACF5;&#xC744; &#xC704;&#xD574; &#xD544;&#xC218;&#xC801; &#xC774;&#xB2E4; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. &#xD568;&#xAED8;, &#xC774;&#xB7EC;&#xD55C; &#xC5F0;&#xAD6C; &#xACB0;&#xACFC; &#xC140; &#xCE68;&#xACF5;&#xC758; &#xC18C;&#xC124; pH &#xC885;&#xC18D; &#xADDC;&#xCE59; &#xC2DD;&#xBCC4;.

Cortactin &#xC778; &#xC0B0;&#xD654; &#xC870;&#xC808; pH &#xC885;&#xC18D; &#xD1B5;&#xB85C; &#xD1B5;&#xD574; &#xC140; &#xCE68;&#xACF5;.

Invadopodios son protuberancias invasivas con actividad proteol&iacute;tica que se encuentra &uacute;nicamente en las c&eacute;lulas tumorales. Fosforilaci&oacute;n de cortactina es un paso clave durante la maduraci&oacute;n de la invadopodios, regular Nck1 vinculante y actividad cofilin. El mecanismo exacto del Reglamento de cofilin de cortactina-dependiente y el papel de esta v&iacute;a en invasi&oacute;n de maduraci&oacute;n y de la c&eacute;lula de invadopodios no se entienden completamente. Proporcionamos evidencia que cortactina-cofilin vinculante est&aacute; regulada por cambios en el pH local en invadopodios que est&aacute;n mediados por el intercambiador sodio hidr&oacute;geno NHE1. Adem&aacute;s, la fosforilaci&oacute;n de la tirosina cortactina media la contrataci&oacute;n de NHE1 del compartimiento de invadopodium, donde localmente aumenta el pH para causar la liberaci&oacute;n de cofilin de cortactina. Demostramos que este mecanismo que implica la fosforilaci&oacute;n cortactina, aumento del pH local y activaci&oacute;n de cofilin regula los ciclos din&aacute;micos de invadopodium saliente y retracci&oacute;n y es esencial para la invasi&oacute;n celular en 3D. Juntos, estos resultados identifican una regulaci&oacute;n de pH-dependent novela de invasi&oacute;n celular.

Fosforilaci&oacute;n de cortactina regula la invasi&oacute;n celular a trav&eacute;s de una v&iacute;a de pH-dependent.

Metastasis requires tumor cell dissemination to different organs from the primary tumor. Dissemination is a complex cell motility phenomenon that requires the molecular coordination of the protrusion, chemotaxis, invasion and contractility activities of tumor cells to achieve directed cell migration. Recent studies of the spatial and temporal activities of the small GTPases have begun to elucidate how this coordination is achieved. The direct visualization of the pathways involved in actin polymerization, invasion and directed migration in dissemination competent tumor cells will help identify the molecular basis of dissemination and allow the design and testing of more specific and selective drugs to block metastasis.

Directed cell invasion and migration during metastasis.

&#x4ECE;&#x539F;&#x53D1;&#x6027;&#x80BF;&#x7624;&#x8F6C;&#x79FB;&#x9700;&#x8981;&#x4E0D;&#x540C;&#x5668;&#x5B98;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x4F20;&#x64AD;&#x3002;&#x4F20;&#x64AD;&#x662F;&#x4E00;&#x79CD;&#x590D;&#x6742;&#x7684;&#x7EC6;&#x80DE;&#x52A8;&#x529B;&#x73B0;&#x8C61;&#xFF0C;&#x9700;&#x8981;&#x5206;&#x5B50;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#xFF0C;&#x4EE5;&#x5B9E;&#x73B0;&#x5B9A;&#x5411;&#x7684;&#x7EC6;&#x80DE;&#x8FC1;&#x79FB;&#x7684;&#x690E;&#x95F4;&#x76D8;&#x7A81;&#x51FA;&#x3001; &#x8D8B;&#x5316;&#x3001; &#x5165;&#x4FB5;&#x548C;&#x6536;&#x7F29;&#x6D3B;&#x52A8;&#x7684;&#x534F;&#x8C03;&#x3002;&#x8FD1;&#x5E74;&#x6765;&#x7814;&#x7A76;&#x7684;&#x7A7A;&#x95F4;&#x548C;&#x65F6;&#x95F4;&#x7684;&#x6D3B;&#x52A8;&#x7684;&#x5C0F;&#x86CB;&#x767D;&#x5DF2;&#x5F00;&#x59CB;&#x6F84;&#x6E05;&#x8FD9;&#x79CD;&#x534F;&#x8C03;&#x5982;&#x4F55;&#x5B9E;&#x73B0;&#x7684;&#x3002;&#x76F4;&#x63A5;&#x53EF;&#x89C6;&#x5316;&#x7684;&#x901A;&#x8DEF;&#x7684;&#x808C;&#x52A8;&#x86CB;&#x767D;&#x805A;&#x5408;&#x3001; &#x5165;&#x4FB5;&#x548C;&#x5B9A;&#x5411;&#x7684;&#x8FC1;&#x79FB;&#x4E2D;&#x4F20;&#x64AD;&#x4E3B;&#x7BA1;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x5C06;&#x5E2E;&#x52A9;&#x786E;&#x5B9A;&#x4F20;&#x64AD;&#x7684;&#x5206;&#x5B50;&#x57FA;&#x7840;&#x548C;&#x5141;&#x8BB8;&#x8BBE;&#x8BA1;&#x548C;&#x6D4B;&#x8BD5;&#x66F4;&#x52A0;&#x5177;&#x4F53;&#x548C;&#x6709;&#x9009;&#x62E9;&#x6027;&#x7684;&#x836F;&#x7269;&#xFF0C;&#x4EE5;&#x963B;&#x6B62;&#x8F6C;&#x79FB;&#x3002;

&#x5B9A;&#x5411;&#x7684;&#x7EC6;&#x80DE;&#x4FB5;&#x88AD;&#x548C;&#x8F6C;&#x79FB;&#x8FC7;&#x7A0B;&#x8FC1;&#x79FB;&#x3002;

M&eacute;tastase exige la diffusion des cellules tumorales &agrave; diff&eacute;rents organes de la tumeur primaire. La diffusion est un ph&eacute;nom&egrave;ne de motilit&eacute; cellulaire complexe qui n&eacute;cessite la coordination mol&eacute;culaire des activit&eacute;s protrusion, chimiotactisme, invasion et la contractilit&eacute; des cellules tumorales pour r&eacute;aliser la migration cellulaire dirig&eacute;e. Des &eacute;tudes r&eacute;centes sur les activit&eacute;s spatiales et temporelles des petites GTPases ont commenc&eacute; &agrave; &eacute;lucider comment cette coordination est r&eacute;alis&eacute;e. La visualisation directe des voies impliqu&eacute;es dans la polym&eacute;risation de l&#39;actine, l&#39;invasion et une migration dirig&eacute;e dans la diss&eacute;mination des cellules tumorales comp&eacute;tente aidera identifier les bases mol&eacute;culaires de la diffusion et permettre la conception et essais de m&eacute;dicaments plus sp&eacute;cifiques et plus s&eacute;lectifs bloquer les m&eacute;tastases.

Invasion des cellules dirig&eacute;es et migration dans le cadre de m&eacute;tastase.

Metastase erfordert Tumor Zelle Verbreitung an verschiedenen Organen aus dem Prim&auml;rtumor. Die Verbreitung ist eine komplexe Zelle Motilit&auml;t-Ph&auml;nomen, das die molekulare Koordinierung der Vorw&ouml;lbung, Chemotaxis, Invasion und Kontraktilit&auml;t Aktivit&auml;ten der Tumorzellen gerichtete Zellmigration zu erreichen erfordert. J&uuml;ngste Studien der r&auml;umlichen und zeitlichen Aktivit&auml;ten des kleinen nun GTPasen aufzukl&auml;ren, wie diese Koordination erreicht wird. Die direkte Visualisierung der Bahnen beteiligt Aktin-Polymerisation, Invasion und gesteuerte Migration in Verbreitung, die zust&auml;ndigen Tumorzellen hilft die molekulare Grundlage der Verbreitung zu identifizieren und erlauben die Gestaltung und Pr&uuml;fung von spezifischer und selektive Drogen Metastasierung zu blockieren.

Gesteuerte Invasion und Migration w&auml;hrend der Metastasierung.

Metastasi richiede la diffusione delle cellule tumorali ad organi diversi dal tumore primario. Diffusione &egrave; un fenomeno di motilit&agrave; cellulare complessa che richiede il coordinamento delle attivit&agrave; protrusione, chemiotassi, invasione e la contrattilit&agrave; delle cellule tumorali per realizzare la migrazione cellulare diretto molecolare. Recenti studi delle attivit&agrave; spaziali e temporali del piccolo GTPases hanno cominciato a spiegare come questo coordinamento &egrave; raggiunto. La visualizzazione diretta delle vie coinvolte nella polimerizzazione dell&#39;actina, invasione e migrazione diretto nella diffusione di cellule tumorali competente verranno aiuter&agrave; a identificare le basi molecolari della diffusione e consentire la progettazione e sperimentazione di farmaci pi&ugrave; specifici e selettivi bloccare le metastasi.

Invasione diretta delle cellule e migrazione durante la metastasi.

&#x8EE2;&#x79FB;&#x304B;&#x3089;&#x539F;&#x767A;&#x816B;&#x760D;&#x306E;&#x5225;&#x306E;&#x81D3;&#x5668;&#x306B;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x306E;&#x666E;&#x53CA;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;&#x3002;&#x666E;&#x53CA;&#x306F;&#x3001;&#x30C7;&#x30A3;&#x30EC;&#x30AF;&#x30C6;&#x30C3;&#x30C9;&#x7D30;&#x80DE;&#x79FB;&#x52D5;&#x3092;&#x9054;&#x6210;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x306E;&#x7A81;&#x8D77;&#x3001;&#x8D70;&#x5316;&#x6027;&#x3001;&#x6D78;&#x6F64;&#x3001;&#x53CE;&#x7E2E;&#x6D3B;&#x52D5;&#x306E;&#x5206;&#x5B50;&#x306E;&#x8ABF;&#x6574;&#x3092;&#x5FC5;&#x8981;&#x3068;&#x3059;&#x308B;&#x8907;&#x96D1;&#x306A;&#x7D30;&#x80DE;&#x904B;&#x52D5;&#x73FE;&#x8C61;&#x3067;&#x3059;&#x3002;&#x6700;&#x8FD1;&#x306E;&#x7814;&#x7A76;&#x3001;&#x5C0F;&#x578B;&#x306E;&#x6642;&#x7A7A;&#x306E;&#x6D3B;&#x52D5;&#x306E;&#x8F38;&#x9001;&#x3053;&#x306E;&#x8ABF;&#x6574;&#x306E;&#x9054;&#x6210;&#x65B9;&#x6CD5;&#x3092;&#x89E3;&#x660E;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x59CB;&#x3081;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x30A2;&#x30AF;&#x30C1;&#x30F3;&#x91CD;&#x5408;&#x3001;&#x4FB5;&#x7565;&#x3001;&#x6709;&#x80FD;&#x306A;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x3092;&#x52A9;&#x3051;&#x308B;&#x666E;&#x53CA;&#x3067;&#x6307;&#x793A;&#x79FB;&#x884C;&#x306B;&#x95A2;&#x4E0E;&#x3059;&#x308B;&#x7D4C;&#x8DEF;&#x306E;&#x76F4;&#x63A5;&#x53EF;&#x8996;&#x5316;&#x666E;&#x53CA;&#x306E;&#x5206;&#x5B50;&#x57FA;&#x76E4;&#x3092;&#x8B58;&#x5225;&#x3057;&#x3001;&#x30C7;&#x30B6;&#x30A4;&#x30F3;&#x3068;&#x3088;&#x308A;&#x5177;&#x4F53;&#x7684;&#x304B;&#x3064;&#x9078;&#x629E;&#x7684;&#x306A;&#x85AC;&#x306E;&#x8A66;&#x9A13;&#x8EE2;&#x79FB;&#x3092;&#x30D6;&#x30ED;&#x30C3;&#x30AF;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;

&#x30C7;&#x30A3;&#x30EC;&#x30AF;&#x30C6;&#x30C3;&#x30C9;&#x7D30;&#x80DE;&#x6D78;&#x6F64;&#x3068;&#x8EE2;&#x79FB;&#x306E;&#x9593;&#x306B;&#x79FB;&#x884C;&#x3002;

&#xC804;&#xC774; &#xAE30;&#xBCF8; &#xC885;&#xC591;&#xC5D0;&#xC11C; &#xB2E4;&#xB978; &#xC7A5;&#xAE30;&#xC5D0; &#xC885;&#xC591; &#xC138;&#xD3EC; &#xBCF4;&#xAE09;&#xC774; &#xD544;&#xC694;&#xD569;&#xB2C8;&#xB2E4;. &#xBCF4;&#xAE09;&#xC740; &#xAC10;&#xB3C5;&#xB41C; &#xC140; &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158; &#xB2EC;&#xC131; &#xD558;&#xAE30; &#xC704;&#xD574; &#xC885;&#xC591; &#xC138;&#xD3EC;&#xC758; &#xB3CC;&#xCD9C;, chemotaxis, &#xCE68;&#xB7B5;, &#xC218;&#xCD95; &#xD65C;&#xB3D9;&#xC758; &#xBD84;&#xC790; &#xC870;&#xC815;&#xC774; &#xD544;&#xC694; &#xBCF5;&#xC7A1; &#xD55C; &#xC138;&#xD3EC;&#xC758; &#xC6B4;&#xB3D9; &#xC131; &#xD604;&#xC0C1;. &#xCD5C;&#xADFC; &#xC5F0;&#xAD6C;&#xB294; &#xC18C;&#xADDC;&#xBAA8;&#xC758; &#xACF5;&#xAC04; &#xBC0F; &#xC2DC;&#xAC04; &#xD65C;&#xB3D9;&#xC758; Gtpases&#xC774;&#xC774; &#xC870;&#xD654;&#xB97C; &#xB2EC;&#xC131; &#xD558;&#xB294; &#xBC29;&#xBC95;&#xC744; &#xBA85;&#xB8CC; &#xD558; &#xAC8C; &#xC2DC;&#xC791; &#xD588;&#xC2B5;&#xB2C8;&#xB2E4;. &#xB9D0;&#xB77C; &#xD569;, &#xCE68;&#xB7B5; &#xBC0F; &#xBCF4;&#xAE09; &#xC720;&#xB2A5;&#xD55C; &#xC885;&#xC591; &#xC138;&#xD3EC; &#xB3C4;&#xC6C0;&#xC774; &#xB420; &#xAC83;&#xC785;&#xB2C8;&#xB2E4; &#xAC10;&#xB3C5;&#xB41C; &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158;&#xC5D0; &#xAD00;&#xB828; &#xB41C; &#xACBD;&#xB85C; &#xC9C1;&#xC811; &#xC2DC;&#xAC01;&#xD654; &#xBCF4;&#xAE09;&#xC758; &#xBD84;&#xC790; &#xAE30;&#xC900;&#xC73C;&#xB85C; &#xC2DD;&#xBCC4; &#xD558; &#xACE0; &#xB514;&#xC790;&#xC778; &#xBC0F; &#xD14C;&#xC2A4;&#xD2B8; &#xB354; &#xAD6C;&#xCCB4;&#xC801;&#xC774; &#xACE0; &#xC120;&#xD0DD;&#xC801; &#xC57D;&#xBB3C;&#xC758; &#xC804;&#xC774; &#xCC28;&#xB2E8; &#xD5C8;&#xC6A9;.

&#xC9C1;&#xC811;&#xB41C; &#xC140; &#xCE68;&#xACF5;&#xACFC; &#xC804;&#xC774; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158;&#xC785;&#xB2C8;&#xB2E4;.

Met&aacute;stasis requiere la difusi&oacute;n de la c&eacute;lula del tumor a distintos &oacute;rganos del tumor primario. La difusi&oacute;n es un fen&oacute;meno de motilidad de la c&eacute;lula compleja que requiere la coordinaci&oacute;n molecular de las actividades saliente, la quimiotaxis, la invasi&oacute;n y la contractilidad de las c&eacute;lulas del tumor para lograr la migraci&oacute;n celular dirigida. Estudios recientes de las actividades espaciales y temporales de las peque&ntilde;as GTPasas han comenzado a dilucidar c&oacute;mo se logra esta coordinaci&oacute;n. La visualizaci&oacute;n directa de las v&iacute;as implicadas en la polimerizaci&oacute;n de la actina, invasi&oacute;n y migraci&oacute;n dirigida en la diseminaci&oacute;n de c&eacute;lulas tumorales competente le ayudar&aacute; a identificar las bases moleculares de la difusi&oacute;n y permite el dise&ntilde;o y pruebas de f&aacute;rmacos m&aacute;s espec&iacute;ficos y selectivos bloquear la met&aacute;stasis.

Invasi&oacute;n celular dirigida y migraci&oacute;n durante la met&aacute;stasis.

Protrusion formation is the first step that precedes cell movement of motile cells. Spatial control of actin polymerization is necessary to achieve directional protrusion during cell migration. Here we show that the spatial coordinators p190RhoGEF and p190RhoGAP regulate actin polymerization during leading edge protrusions by regulating the actin barbed end distribution and amplitude. The distribution of RhoC activity and proper balance of cofilin activation achieved by p190RhoGEF and p190RhoGAP determines the direction of final protrusive activity. These findings provide a new insight into the dynamic plasticity in the amplitude and distribution of barbed ends, which can be modulated by fine-tuning RhoC activity by upstream GEFs and GAPs for directed cell motility.

Spatial regulation of RhoC activity defines protrusion formation in migrating cells.

Recently, a consensus has emerged that cofilin severing activity can generate free actin filament ends that are accessible for F-actin polymerization and depolymerization without changing the rate of G-actin association and dissociation at either filament end. The structural basis of actin filament severing by cofilin is now better understood. These results have been integrated with recently discovered mechanisms for cofilin activation in migrating cells, which led to new models for cofilin function that provide insights into how cofilin regulation determines the temporal and spatial control of cell behaviour.

Functions of cofilin in cell locomotion and invasion.

Tumor cell motility and invasion rely on actin cytoskeleton rearrangements mediated by the activation of RhoGTPase signaling pathways. Invadopodia are membrane-degrading protrusions that mediate extracellular matrix degradation. Here, we provide procedures for imaging RhoGTPase biosensors in tumor cells during the formation of invadopodia and matrix degradation.

Live cell imaging of RhoGTPase biosensors in tumor cells.

Biosensors based on FRET have been useful in deciphering the dynamics of protein activation events in living cells at subcellular resolutions and in time scales of seconds. These new systems allow observations of dynamic processes which were not possible previously using more traditional biochemical and cell biological approaches. The image data sets obtained from these sensors require careful processing in order to represent the actual protein activation events. Here, we will cover the basic approaches useful for processing the raw image data sets into relativistic ratiometric measurements, capable of depicting relative differences in the protein activation states within a single cell. We will discuss in detail the approaches for genetically encoded, single-chain biosensor systems based on FRET, as well as those that are based on intermolecular, dual-chain design. Additionally, the same analysis can be utilized for biosensor systems using solvatochromic dyes (Nalbant, Hodgson, Kraynov, Toutchkine, & Hahn, 2004), useful for detection of endogenous protein activation states.

Quantitative ratiometric imaging of FRET-biosensors in living cells.

Systemic metastasis is the dissemination of cancer cells from the primary tumor to distant organs and is the primary cause of death in cancer patients. How do cancer cells leave the primary tumor mass? The ability of the tumor cells to form different types of actin-rich protrusions including invasive protrusions (invadopodia) and locomotory protrusions (lamellipodia [2D] or pseudopodia [3D]), facilitate the invasion and dissemination of the tumor cells. Rho-family of p21 small GTPases plays a direct role in regulating the actin dynamics in these intracellular compartments. Recent studies have shown that the signaling molecules including RhoC/p190RhoGEF/p190RhoGAP acts as a "molecular compass" in order to direct the spatial and temporal dynamics of the formation of these invasive and locomotory protrusions leading to efficient invasion.

Spatial regulation of tumor cell protrusions by RhoC.

Rho family GTPases control cell migration and participate in the regulation of cancer metastasis. Invadopodia, associated with invasive tumour cells, are crucial for cellular invasion and metastasis. To study Rac1 GTPase in invadopodia dynamics, we developed a genetically encoded, single-chain Rac1 fluorescence resonance energy (FRET) transfer biosensor. The biosensor shows Rac1 activity exclusion from the core of invadopodia, and higher activity when invadopodia disappear, suggesting that reduced Rac1 activity is necessary for their stability, and Rac1 activation is involved in disassembly. Photoactivating Rac1 at invadopodia confirmed this previously unknown Rac1 function. We describe here an invadopodia disassembly model, where a signalling axis involving TrioGEF, Rac1, Pak1, and phosphorylation of cortactin, causes invadopodia dissolution. This mechanism is critical for the proper turnover of invasive structures during tumour cell invasion, where a balance of proteolytic activity and locomotory protrusions must be carefully coordinated to achieve a maximally invasive phenotype.

A Trio-Rac1-Pak1 signalling axis drives invadopodia disassembly.

Invadopodia are actin-rich protrusions that degrade the extracellular matrix and are required for stromal invasion, intravasation, and metastasis. The role of the focal adhesion protein talin in regulating these structures is not known. Here, we demonstrate that talin is required for invadopodial matrix degradation and three-dimensional extracellular matrix invasion in metastatic breast cancer cells. The sodium/hydrogen exchanger 1 (NHE-1) is linked to the cytoskeleton by ezrin/radixin/moesin family proteins and is known to regulate invadopodium-mediated matrix degradation. We show that the talin C terminus binds directly to the moesin band 4.1 ERM (FERM) domain to recruit a moesin-NHE-1 complex to invadopodia. Silencing talin resulted in a decrease in cytosolic pH at invadopodia and blocked cofilin-dependent actin polymerization, leading to impaired invadopodium stability and matrix degradation. Furthermore, talin is required for mammary tumor cell motility, intravasation, and spontaneous lung metastasis in vivo. Thus, our findings provide a novel understanding of how intracellular pH is regulated and a molecular mechanism by which talin enhances tumor cell invasion and metastasis.

Talin regulates moesin-NHE-1 recruitment to invadopodia and promotes mammary tumor metastasis.

Metastasis is the main cause of death in breast cancer patients. Cell migration is an essential component of almost every step of the metastatic cascade, especially the early step of invasion inside the primary tumor. In this report, we have used intravital multiphoton microscopy to visualize the different migration patterns of human breast tumor cells in live primary tumors. We used xenograft tumors of MDA-MB-231 cells as well as a low passage xenograft tumor from orthotopically injected patient-derived breast tumor cells. Direct visualization of human tumor cells in vivo shows two patterns of high-speed migration inside primary tumors: (1) single cells and (2) multicellular streams (i.e., cells following each other in a single file but without cohesive cell junctions). Critically, we found that only streaming and not random migration of single cells was significantly correlated with proximity to vessels, with intravasation and with numbers of elevated circulating tumor cells in the bloodstream. Finally, although the two human tumors were derived from diverse genetic backgrounds, we found that their migratory tumor cells exhibited coordinated gene expression changes that led to the same end-phenotype of enhanced migration involving activating actin polymerization and myosin contraction. Our data are the first direct visualization and assessment of in vivo migration within a live patient-derived breast xenograft tumor.

Intravital multiphoton imaging reveals multicellular streaming as a crucial component of in vivo cell migration in human breast tumors.

The Rho-family of p21 small GTPases are directly linked to the regulation of actin-based motile machinery and play a key role in the control of cell migration. Aside from the original and most well-characterized canonical Rho GTPases RhoA, Rac1, and Cdc42, numerous isoforms of these key proteins have been identified and shown to have specific roles in regulating various cellular motility processes. The major difficulty in addressing these isoform-specific effects is that isoforms typically contain highly similar primary amino acid sequences and thus are able to interact with the same upstream regulators and the downstream effector targets. Here, we will introduce the major members of each GTPase subfamily and discuss recent advances in the design and application of fluorescent resonance energy transfer-based probes, which are at the forefront of the technologies available to directly probe the differential, spatiotemporal activation dynamics of these proteins in live single cells. Currently, it is possible to specifically detect the activation status of RhoA vs. RhoC isoforms, as well as Cdc42 vs. TC-10 isoforms in living cells. Clearly, additional efforts are still required to produce biosensor systems capable of detecting other isoforms of Rho GTPases including RhoB, Rac2/3, RhoG, etc. Through such efforts, we will uncover the isoform-specific roles of these near-identical proteins in living cells, clearly an important area of the Rho GTPase biology that is not yet fully appreciated.

Rho GTPase isoforms in cell motility: Don't fret, we have FRET.

The process of intravasation involving transendothelial migration is a key step in metastatic spread. How the triple cell complex composed of a macrophage, Mena over-expressing tumor cell and endothelial cell, called the tumor microenvironment of metastasis (TMEM), facilitates tumor cell transendothelial migration is not completely understood. Previous work has shown that the physical contact between a macrophage and tumor cell results in the formation of invadopodia, actin-rich matrix degrading protrusions, important for tumor cell invasion and transendothelial migration and tumor cell dissemination. Herein, we show that the macrophage-induced invadopodium is formed through a Notch1/Mena(INV) signaling pathway in the tumor cell upon macrophage contact. This heterotypic tumor cell - macrophage interaction results in the upregulation of Mena(INV) through the activation of MENA transcription. Notch1 and Mena(INV) expression are required for tumor cell transendothelial migration, a necessary step during intravasation. Inhibition of the Notch signaling pathway blocked macrophage-induced invadopodium formation in vitro and the dissemination of tumor cells from the primary tumor in vivo. Our findings indicate a novel role for Notch1 signaling in the regulation of Mena(INV) expression and transendothelial migration and provide mechanistic information essential to the use of therapeutic inhibitors of metastasis.

Macrophage-dependent tumor cell transendothelial migration is mediated by Notch1/Mena(INV)-initiated invadopodium formation.

Hypoxia is a poor-prognosis microenvironmental hallmark of solid tumours, but it is unclear how it influences the fate of disseminated tumour cells (DTCs) in target organs. Here we report that hypoxic HNSCC and breast primary tumour microenvironments displayed upregulation of key dormancy (NR2F1, DEC2, p27) and hypoxia (GLUT1, HIF1α) genes. Analysis of solitary DTCs in PDX and transgenic mice revealed that post-hypoxic DTCs were frequently NR2F1(hi)/DEC2(hi)/p27(hi)/TGFβ2(hi) and dormant. NR2F1 and HIF1α were required for p27 induction in post-hypoxic dormant DTCs, but these DTCs did not display GLUT1(hi) expression. Post-hypoxic DTCs evaded chemotherapy and, unlike ER(-) breast cancer cells, post-hypoxic ER(+) breast cancer cells were more prone to enter NR2F1-dependent dormancy. We propose that primary tumour hypoxic microenvironments give rise to a subpopulation of dormant DTCs that evade therapy. These post-hypoxic dormant DTCs may be the source of disease relapse and poor prognosis associated with hypoxia.

Article	Year
Novel lambda FRET spectral confocal microscopy imaging method. Microscopy research and technique\| PubMed ID: 18785251	2009
Specific tyrosine phosphorylation sites on cortactin regulate Nck1-dependent actin polymerization in invadopodia. Journal of cell science\| PubMed ID: 20971703	2010
An EGFR-Src-Arg-cortactin pathway mediates functional maturation of invadopodia and breast cancer cell invasion. Cancer research\| PubMed ID: 21257711	2011
A novel spatiotemporal RhoC activation pathway locally regulates cofilin activity at invadopodia. Current biology : CB\| PubMed ID: 21474314	2011
Cortactin phosphorylation regulates cell invasion through a pH-dependent pathway. The Journal of cell biology\| PubMed ID: 22105349	2011
Directed cell invasion and migration during metastasis. Current opinion in cell biology\| PubMed ID: 22209238	2011
Spatial regulation of RhoC activity defines protrusion formation in migrating cells. Journal of cell science\| PubMed ID: 23704350	2013
Functions of cofilin in cell locomotion and invasion. Nature reviews. Molecular cell biology\| PubMed ID: 23778968	2013
Live cell imaging of RhoGTPase biosensors in tumor cells. Methods in molecular biology (Clifton, N.J.)\| PubMed ID: 23868600	2013
Quantitative ratiometric imaging of FRET-biosensors in living cells. Methods in cell biology\| PubMed ID: 23931524	2013
Spatial regulation of tumor cell protrusions by RhoC. Cell adhesion & migration\| PubMed ID: 24642482	2014
A Trio-Rac1-Pak1 signalling axis drives invadopodia disassembly. Nature cell biology\| PubMed ID: 24859002	2014
Talin regulates moesin-NHE-1 recruitment to invadopodia and promotes mammary tumor metastasis. The Journal of cell biology\| PubMed ID: 24891603	2014
Intravital multiphoton imaging reveals multicellular streaming as a crucial component of in vivo cell migration in human breast tumors. Intravital\| PubMed ID: 25013744	2013
Rho GTPase isoforms in cell motility: Don't fret, we have FRET. Cell adhesion & migration\| PubMed ID: 25482645	2014
A Trio-Rac1-Pak1 signalling axis drives invadopodia disassembly. Nature cell biology\| PubMed ID: 25720965	2015
Macrophage-dependent tumor cell transendothelial migration is mediated by Notch1/Mena(INV)-initiated invadopodium formation. Scientific reports\| PubMed ID: 27901093	2016
Phenotypic heterogeneity of disseminated tumour cells is preset by primary tumour hypoxic microenvironments. Nature cell biology\| PubMed ID: 28114271	2017

Jose Javier Bravo-Cordero

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