in-vivo-detection-analysis-rb-protein-sumoylation-human

<em>In Vivo</em> Detection and Analysis of Rb Protein SUMOylation in Human Cells

The post-translational modifications of proteins are critical for the proper regulation of intracellular signal transduction. Among these modifications, ...

Eye and ENT Hospital of Fudan University

Fudan University

Th-cytokines play important roles in dacryoadenitis associated with Sjögren's syndrome (SS). The aims of this study were to (i) investigate whether interleukin (IL)-17, a newly identified Th-cytokine, participates in autoimmune dacryoadenitis of MRL/lpr mice and (ii) examine the relationships among IL-17, IL-4, and interferon (IFN)-γ in these mice.

Expression of interleukin-17 in autoimmune dacryoadenitis in MRL/lpr mice.

&#x4EE5;&#x524D;&#x7684;&#x7814;&#x7A76;&#x8868;&#x660E;&#x4EBA;&#x53E3;&#x5C11;&#x7684;&#x80BF;&#x7624;&#x5E72;&#x7EC6;&#x80DE;&#x6837;&#x7EC6;&#x80DE;&#x5B58;&#x5728;&#x89C6;&#x7F51;&#x819C;&#x6BCD;&#x7EC6;&#x80DE;&#x7624;&#x3002;&#x4E3A;&#x7814;&#x7A76;&#x8FD9;&#x4E00;&#x4EBA;&#x7FA4;&#x63D0;&#x4F9B;&#x6A21;&#x578B;&#xFF0C;&#x6211;&#x4EEC;&#x5BFB;&#x6C42;&#x5EFA;&#x7ACB;&#x4E00;&#x4E2A;&#x957F;&#x671F;&#x7684;&#x6587;&#x5316;&#x4ECE;&#x4EBA;&#x7C7B;&#x89C6;&#x7F51;&#x819C;&#x6BCD;&#x7EC6;&#x80DE;&#x7624;&#x5177;&#x6709;&#x764C;&#x75C7;&#x5E72;&#x7EC6;&#x80DE;&#x6837;&#x5C5E;&#x6027;&#x3002;&#x65B0;&#x9C9C;&#x80BF;&#x7624;&#x7EC4;&#x7EC7;&#x4E86;&#x6D88;&#x5316;&#x548C;&#x65E0;&#x8840;&#x6E05;&#x57F9;&#x517B;&#x57FA;&#x4E2D;&#x57F9;&#x517B;&#x3002;&#x80BF;&#x7624;&#x9886;&#x57DF;&#x5F62;&#x6210;&#x548C;&#x4E0D;&#x65AD;&#x88AB; passaged&#x3002;&#x5E72;&#x7EC6;&#x80DE;&#x5C5E;&#x6027;&#x901A;&#x8FC7;&#x6807;&#x672C;&#x3001; &#x5B9E;&#x65F6;&#x5B9A;&#x91CF;&#x9006;&#x8F6C;&#x5F55;-&#x805A;&#x5408;&#x9176;&#x94FE;&#x53CD;&#x5E94;&#x548C;&#x5316;&#x7597;&#x8010;&#x836F;&#x68C0;&#x6D4B;&#x8FDB;&#x884C;&#x4E86;&#x5BA1;&#x67E5;&#x3002;&#x81F4;&#x7624;&#x6027;&#x80BF;&#x7624;&#x9886;&#x57DF;&#x5F62;&#x6210;&#x7EC6;&#x80DE;&#x5F02;&#x79CD;&#x79FB;&#x690D;&#x5B9E;&#x9A8C;&#x6240;&#x8BC1;&#x5B9E;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x6211;&#x4EEC;&#x5BA1;&#x67E5;&#x4E86;&#x7EC6;&#x80DE;&#x8868;&#x9762;&#x6807;&#x8BB0;&#x7269; CD133 &#x548C; CD44 &#x7684;&#x8868;&#x8FBE;&#x3002;&#x80BF;&#x7624;&#x7EC6;&#x80DE;&#x4F5C;&#x4E3A;&#x6D6E;&#x52A8;&#x8D85;&#x8FC7; 30 &#x901A;&#x9053;&#x9886;&#x57DF;&#x6269;&#x5927;&#x3002;&#x9886;&#x57DF;&#x5F62;&#x6210;&#x7EC6;&#x80DE;&#x4ECB;&#x5BFC; Oct&#x2011;4&#x3001; &#x5DE2;&#x86CB;&#x767D;&#x548C; Pax6 &#x5E72;&#x7EC6;&#x80DE;&#x7684;&#x57FA;&#x56E0;&#x3002;&#x86CB;&#x767D;&#x514D;&#x75AB;&#x67D3;&#x8272;&#x7684;&#x9886;&#x57DF;&#x5DE2;&#x86CB;&#x767D;&#x3001; Pax6&#xFF0C;&#x548C; ABCG2 &#x663E;&#x793A;&#x9633;&#x6027;&#x3002;&#x76F8;&#x6BD4;&#x4E4B;&#x4E0B;&#xFF0C;&#x5206;&#x5316;&#x7684;&#x7EC6;&#x80DE;&#x6765;&#x81EA;&#x6210;&#x719F;&#x7684;&#x89C6;&#x7F51;&#x819C;&#x7EC6;&#x80DE;&#x6807;&#x8BB0;&#x7269; MAP2&#xFF0C;&#x80F6;&#x8D28;&#x7EA4;&#x7EF4;&#x9178;&#x6027;&#x86CB;&#x767D;&#xFF0C;recoverin&#xFF0C;&#x7EC6;&#x83CC;&#x89C6;&#x7D2B;&#x7EA2;&#x8D28; B &#x548C;&#x6D77;&#x519B;&#x7814;&#x7A76;&#x5B9E;&#x9A8C;&#x6240;&#xFF0C;&#x4EE5;&#x53CA;&#x663E;&#x793A;&#x514D;&#x75AB;&#x53CD;&#x5E94;&#x6027;&#x7684; NF200&#xFF0C;&#x80F6;&#x8D28;&#x7EA4;&#x7EF4;&#x9178;&#x6027;&#x86CB;&#x767D;&#xFF0C;recoverin &#x548C; PKC&alpha; &#x7684;&#x8FD9;&#x4E9B;&#x9886;&#x57DF;&#x8868;&#x793A;&#x9AD8;&#x7EA7;&#x522B;&#x3002;&#x6B64;&#x5916;&#xFF0C;CD44 &#x548C; CD133 &#x9AD8;&#x5EA6;&#x8868;&#x793A;&#x5728;&#x9886;&#x57DF;&#x5F62;&#x6210;&#x4E0E;&#x5206;&#x5316;&#x7EC6;&#x80DE;&#x7684;&#x7EC6;&#x80DE;&#x3002;&#x9886;&#x57DF;&#x5F62;&#x6210;&#x7EC6;&#x80DE;&#x5411;&#x5361;&#x94C2;&#x4E0E;&#x5206;&#x5316;&#x7684;&#x7EC6;&#x80DE;&#x663E;&#x793A;&#x8F83;&#x9AD8;&#x7684;&#x5316;&#x7597;&#x8010;&#x836F;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x4EBA;&#x5DE5;&#x6676;&#x72B6;&#x4F53;&#x7684;&#x5C11;&#x5219; 2 x 103 &#x9886;&#x57DF;&#x5F62;&#x6210;&#x7EC6;&#x80DE;&#x6CE8;&#x5165; NOD/SCID &#x5C0F;&#x9F20;&#x5F15;&#x8D77;&#x7C7B;&#x4F3C;&#x4E8E;&#x539F;&#x59CB;&#x7684;&#x75C5;&#x4EBA;&#x80BF;&#x7624;&#x7684;&#x65B0;&#x80BF;&#x7624;&#x3002;&#x8FD9;&#x4E9B;&#x7ED3;&#x679C;&#x663E;&#x793A;&#x9886;&#x57DF;&#x5F62;&#x6210;&#x7684;&#x5355;&#x5143;&#x683C;&#x4FDD;&#x7559;&#x5176;&#x5E72;&#x7EC6;&#x80DE;&#x7684;&#x5C5E;&#x6027;&#x548C;&#x81F4;&#x7624;&#x6027;&#xFF0C;&#x751A;&#x81F3;&#x540E;&#x957F;&#x671F;&#x57F9;&#x517B;&#x3002;&#x8FD9;&#x5C06;&#x662F;&#x5408;&#x9002;&#x7684;&#x4F53;&#x5916;&#x6A21;&#x578B;&#x5728;&#x89C6;&#x7F51;&#x819C;&#x6BCD;&#x7EC6;&#x80DE;&#x7624;&#x80BF;&#x7624;&#x5E72;&#x7EC6;&#x80DE;&#x7684;&#x7814;&#x7A76;&#xFF0C;&#x5E76;&#x5236;&#x5B9A;&#x5316;&#x7597;&#x836F;&#x7269;&#x548C;&#x6218;&#x7565;&#x3002;

&#x901A;&#x8FC7;&#x4ECE;&#x89C6;&#x7F51;&#x819C;&#x6BCD;&#x7EC6;&#x80DE;&#x7624;&#x7684;&#x957F;&#x671F;&#x65E0;&#x8840;&#x6E05;&#x57F9;&#x517B;&#x89C6;&#x7F51;&#x819C;&#x764C;&#x75C7;&#x5E72;&#x7EC6;&#x80DE;&#x6837;&#x5C5E;&#x6027;&#x7684;&#x7EF4;&#x62A4;&#x3002;

Previous studies have demonstrated that a small population of cancer stem cell-like cells exists in retinoblastoma. To provide a model for studying this population, we sought to establish a long-term culture from human retinoblastoma that have cancer stem cell-like properties. Fresh tumor tissue was digested and cultured in serum-free medium. Tumor spheres formed and were passaged continuously. Stem cell properties were examined through immunostaining, real-time quantitative RT-PCR and chemoresistance assay. Tumorigenicity of the tumor sphere-forming cells was confirmed by xenograft experiments. Furthermore, we examined the expression of cell surface markers CD44 and CD133. Tumor cells expanded as floating spheres for more than 30 passages. Sphere-forming cells overexpressed stem cell genes Octâ€‘4, Nestin and Pax6. Immunostaining of spheres showed positivity for Nestin, Pax6 and also ABCG2. In contrast, differentiated cells derived from these spheres expressed high levels of mature retinal cell markers MAP2, GFAP, recoverin, Opsin B and Nrl, and showed immunoreactivity for NF200, GFAP, recoverin and PKCÎ±. Furthermore, both CD44 and CD133 were highly expressed in sphere-forming cells vs. differentiated cells. Sphere-forming cells displayed higher chemoresistance to carboplatin as opposed to differentiated cells. Moreover, intraocular injection of as few as 2x103 sphere-forming cells into NOD/SCID mice gave rise to new tumors similar to the original patient tumors. These results revealed that the sphere-forming cells preserved their stem cell properties and tumorigenicity, even after long-term culture. This would be a suitable inÂ vitro model to study cancer stem-like cells in retinoblastoma and to develop chemotherapeutic drugs and strategies.

Maintenance of retinal cancer stem cell-like properties through long-term serum-free culture from human retinoblastoma.

Des &eacute;tudes ant&eacute;rieures ont d&eacute;montr&eacute; qu&#39;une petite population de cellules semblables &agrave; des cellules souches du cancer existe dans le r&eacute;tinoblastome. Pour fournir un mod&egrave;le pour l&#39;&eacute;tude de cette population, nous avons cherch&eacute; &agrave; &eacute;tablir une culture &agrave; long terme de r&eacute;tinoblastome humaine ayant des propri&eacute;t&eacute;s de cellules souches du cancer. Tissu tumoral frais a &eacute;t&eacute; dig&eacute;r&eacute; et cultiv&eacute; dans un milieu sans s&eacute;rum. Tumeur sph&egrave;res forment et ont &eacute;t&eacute; repiqu&eacute;es en permanence. Propri&eacute;t&eacute;s de cellules souches ont &eacute;t&eacute; examin&eacute;es par immunomarquage, RT-PCR quantitative en temps r&eacute;el et dosage de la chimior&eacute;sistance. Tumorig&eacute;nicit&eacute; des cellules tumorales formant sph&egrave;re a &eacute;t&eacute; confirm&eacute;e par des exp&eacute;riences de x&eacute;nogreffes. En outre, nous avons &eacute;tudi&eacute; l&#39;expression des marqueurs de surface de cellules CD44 et CD133. Les cellules tumorales &eacute;largi comme flottant les sph&egrave;res pour plus de 30 passages. Cellules h&eacute;matopo&iuml;&eacute;tiques sph&egrave;re surexprim&eacute; g&egrave;nes de cellules souches Oct&#x2011;4, Nestin et Pax6. Immunostaining des sph&egrave;res ont montr&eacute; la positivit&eacute; de Nestin, Pax6 et aussi ABCG2. En revanche, diff&eacute;renci&eacute;s de cellules d&eacute;riv&eacute;es de ces sph&egrave;res ont exprim&eacute; des niveaux &eacute;lev&eacute;s des marqueurs de cellules r&eacute;tiniennes mature carte2, GFAP, recoverin, opsine B et Nrl et a montr&eacute; l&#39;immunor&eacute;activit&eacute; pour NF200, GFAP, recoverin et PKC&alpha;. En outre, CD44 tant CD133 &eacute;taient fortement exprim&eacute;e dans la sph&egrave;re formant les cellules contre les cellules diff&eacute;renci&eacute;es. Cellules h&eacute;matopo&iuml;&eacute;tiques sph&egrave;re affichent chimior&eacute;sistance sup&eacute;rieur au carboplatine par opposition aux cellules diff&eacute;renci&eacute;es. Par ailleurs, l&#39;injection intraoculaire d&#39;aussi peu que 2 x 103 cellules de formation sph&egrave;re dans la souris NOD/SCID a donn&eacute; lieu &agrave; nouvelles tumeurs semblables pour les tumeurs de patients originales. Ces r&eacute;sultats ont r&eacute;v&eacute;l&eacute; que les cellules formant des sph&egrave;re conserv&eacute; leurs propri&eacute;t&eacute;s de cellules souches et de la tumorig&eacute;nicit&eacute;, m&ecirc;me apr&egrave;s les cultures &agrave; long terme. Ce serait un mod&egrave;le in vitro appropri&eacute; pour &eacute;tudier les cellules canc&eacute;reuses de souches ressemblant au r&eacute;tinoblastome et &eacute;laborer des strat&eacute;gies et des m&eacute;dicaments chimioth&eacute;rapeutiques.

Entretien de propri&eacute;t&eacute;s cellules souches r&eacute;tiniennes cancer par le biais de la culture sans s&eacute;rum &agrave; long terme de r&eacute;tinoblastome humaine.

Fr&uuml;here Studien haben bewiesen, dass eine kleine Population von Krebszellen Stammzell-wie in einem Retinoblastom vorhanden ist. Um ein Modell f&uuml;r die Untersuchung dieser Bev&ouml;lkerung zu erm&ouml;glichen, wollte wir einf&uuml;hren einer Langzeitkultur von menschlichen Retinoblastom, die Krebs Stammzell-&auml;hnliche Eigenschaften aufweisen. Frische Tumorgewebe wurde verdaut und in serumfreien Medium kultiviert. Tumor-Kugeln gebildet und wurden kontinuierlich passaged. Stammzell-Eigenschaften wurden durch Immunf&auml;rbung, Echtzeit-quantitative RT-PCR und Chemoresistance-Assay untersucht. Tumorigenicity der Tumor Kugel-bildenden Zellen wurde durch Xenograft--Experimente best&auml;tigt. Dar&uuml;ber hinaus haben wir den Ausdruck der Zelle Oberfl&auml;che Marker CD44 und CD133 gepr&uuml;ft. Tumorzellen erweitert als schwimmende Kugeln f&uuml;r mehr als 30 Passagen. Kugel-bildenden Zellen Pleuramesotheliom Stammzellen Gene Oct&#x2011;4, Nestin und Pax6. Paraffinschnitten Sph&auml;ren zeigte Positivit&auml;t f&uuml;r Nestin, Pax6 und auch ABCG2. Dagegen differenzierte Zellen aus diesen Kugeln ausgedr&uuml;ckt hohe reifer retinale Zell-Markern MAP2, GFAP, recoverin, Opsin B und Nrl und zeigte Immunoreactivity f&uuml;r NF200, GFAP, recoverin und PKC&alpha;. Dar&uuml;ber hinaus CD44 und CD133 waren hoch ausgedr&uuml;ckt in Kugel-bildenden Zellen vs. differenzierte Zellen. Kugel-bildenden Zellen h&ouml;herer Chemoresistance, Carboplatin im Gegensatz zu differenzierten Zellen angezeigt. Dar&uuml;ber hinaus brachte intraokulare Injektion von nur 2 x 103 Kugel-bildenden Zellen in NOD/SCID M&auml;use neue Tumoren, die &auml;hnlich wie die urspr&uuml;nglichen Patienten Tumoren. Diese Ergebnisse ergaben, dass die Kugel-bildenden Zellen ihre Stammzellen-Eigenschaften und Tumorigenicity, auch nach Langzeitkultur gew&auml;hrleistet. Dies w&auml;re ein geeignetes in-vitro-Modell Vorbau-wie Krebszellen im Retinoblastom zu studieren und Chemotherapeutika und Strategien zu entwickeln.

Wartung der Netzhaut Krebs Stammzell-&auml;hnliche Eigenschaften durch serumfreie Langzeitkultur von menschlichen Retinoblastom.

Precedenti studi hanno dimostrato che una piccola popolazione di cellule staminali-come le cellule tumorali presente nel retinoblastoma. Per fornire un modello per lo studio di questa popolazione, abbiamo cercato di creare una cultura a lungo termine da retinoblastoma umano che hanno il cancro delle cellule staminali-come propriet&agrave;. Tessuto tumorale fresco &egrave; stato digerito e coltivato in terreno privo di siero. Sfere di tumore formano e sono state attraversate continuamente. Propriet&agrave; delle cellule staminali sono state esaminate tramite immunostaining, RT-PCR quantitativa in tempo reale e analisi chemoresistance. Tumorigenicity di sfera-formare le cellule del tumore &egrave; stata confermata da esperimenti xenoinnesto. Inoltre, abbiamo esaminato l&#39;espressione di marcatori di superficie delle cellule CD44 e CD133. Le cellule tumorali ampliato come galleggianti sfere per pi&ugrave; di 30 passaggi. Cellule che formano sfera overexpressed geni di cellule staminali Oct&#x2011;4, Nestin e Pax6. Immunostaining delle sfere ha mostrato positivit&agrave; per Nestin, Pax6 e anche ABCG2. Al contrario, differenziato cellule derivate da questi sfere espresse alti livelli di marcatori di cellule retiniche maturo MAP2, GFAP, recoverina, Opsin B e Nrl e ha mostrato immunoreattivit&agrave; per NF200, GFAP, recoverina e PKC&alpha;. Inoltre, entrambi CD44 e CD133 erano altamente espresso in sfera-formare le cellule vs cellule differenziate. Cellule che formano sfera visualizzato chemoresistance superiore a carboplatino rispetto a cellule differenziate. Inoltre, iniezione intraoculare di come pochi come cellule che formano sfera 2 x 103 in topi NOD/SCID diede origine a nuovi tumori simili a tumori pazienti originali. Questi risultati hanno rivelato che le cellule che formano sfera conservato loro tumorigenicity e le propriet&agrave; delle cellule staminali anche dopo coltura a lungo termine. Questo sarebbe un adatto modello in vitro per lo studio di cellule simil-staminali tumorali nel retinoblastoma e sviluppare strategie e farmaci chemioterapici.

Manutenzione delle propriet&agrave; delle cellule staminali-come cancro della retina attraverso la cultura a lungo termine serum-free da retinoblastoma umano.

&#x524D;&#x306E;&#x7814;&#x7A76;&#x306F;&#x3001;&#x7DB2;&#x819C;&#x82BD;&#x7D30;&#x80DE;&#x816B;&#x306B;&#x304A;&#x3051;&#x308B;&#x764C;&#x5E79;&#x7D30;&#x80DE;&#x69D8;&#x7D30;&#x80DE;&#x306E;&#x5C0F;&#x96C6;&#x56E3;&#x304C;&#x5B58;&#x5728;&#x3059;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x4EBA;&#x53E3;&#x3092;&#x52C9;&#x5F37;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;&#x30E2;&#x30C7;&#x30EB;&#x3092;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x306B;&#x306F;&#x3001;&#x304C;&#x3093;&#x5E79;&#x7D30;&#x80DE;&#x306E;&#x3088;&#x3046;&#x306A;&#x30D7;&#x30ED;&#x30D1;&#x30C6;&#x30A3;&#x3092;&#x6301;&#x3064;&#x4EBA;&#x9593;&#x306E;&#x7DB2;&#x819C;&#x82BD;&#x816B;&#x304B;&#x3089;&#x306E;&#x9577;&#x671F;&#x7684;&#x306A;&#x6587;&#x5316;&#x3092;&#x78BA;&#x7ACB;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x6C42;&#x3081;&#x305F;&#x3002;&#x65B0;&#x9BAE;&#x306A;&#x816B;&#x760D;&#x7D44;&#x7E54;&#x306F;&#x6D88;&#x5316;&#x3057;&#x3001;&#x7121;&#x8840;&#x6E05;&#x57F9;&#x5730;&#x3067;&#x57F9;&#x990A;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x816B;&#x760D;&#x306E;&#x7403;&#x5F62;&#x6210;&#x304A;&#x3088;&#x3073;&#x9023;&#x7D9A;&#x7D99;&#x4EE3;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x5E79;&#x7D30;&#x80DE;&#x7279;&#x6027;&#x306F;&#x514D;&#x75AB;&#x67D3;&#x8272;, &#x30EA;&#x30A2;&#x30EB;&#x30BF;&#x30A4;&#x30E0;&#x5B9A;&#x91CF;&#x7684; RT-PCR &#x304A;&#x3088;&#x3073; chemoresistance &#x3092;&#x7570;&#x306A;&#x308A;&#x30A2;&#x30C3;&#x30BB;&#x30A4;&#x3092;&#x901A;&#x3058;&#x3066;&#x3092;&#x8ABF;&#x3079;&#x305F;&#x3002;17. &#x7403;&#x5F62;&#x6210;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x7570;&#x7A2E;&#x79FB;&#x690D;&#x5B9F;&#x9A13;&#x306B;&#x3088;&#x3063;&#x3066;&#x78BA;&#x8A8D;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x30BB;&#x30EB;&#x8868;&#x9762;&#x306E;&#x30DE;&#x30FC;&#x30AB;&#x30FC; CD44 &#x3068; CD133 &#x306E;&#x767A;&#x73FE;&#x3092;&#x691C;&#x8A0E;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x816B;&#x760D;&#x7D30;&#x80DE;&#x306F; 30 &#x4EE5;&#x4E0A;&#x306E;&#x901A;&#x8DEF;&#x306E;&#x305F;&#x3081;&#x306E;&#x7403;&#x3092;&#x6D6E;&#x52D5;&#x3068;&#x3057;&#x3066;&#x62E1;&#x5927;&#x3002;&#x7403;&#x5F62;&#x6210;&#x7D30;&#x80DE; Oct&#x2011;4&#x3001;Nestin&#x3001;Pax6 &#x306E;&#x830E;&#x7D30;&#x80DE;&#x306E;&#x907A;&#x4F1D;&#x5B50;&#x3092;&#x904E;&#x5270;&#x767A;&#x73FE;&#x3002;&#x7403;&#x306E;&#x514D;&#x75AB;&#x67D3;&#x8272;&#x967D;&#x6027; Nestin&#x3001;Pax6 &#x3068;&#x307E;&#x305F; ABCG2 &#x3092;&#x793A;&#x3057;&#x305F;&#x3002;&#x5BFE;&#x7167;&#x7684;&#x306B;&#x3001;&#x6210;&#x719F;&#x3057;&#x305F;&#x7DB2;&#x819C;&#x7D30;&#x80DE;&#x30DE;&#x30FC;&#x30AB;&#x30FC; MAP2, GFAP&#x3001;recoverin&#x3001;&#x30AA;&#x30D7;&#x30B7;&#x30F3; B &#x3068; Nrl &#x3068; NF200, GFAP&#x3001;recoverin &#x306E;&#x53CD;&#x5FDC;&#x3068; PKC&alpha; &#x306E;&#x3053;&#x308C;&#x3089;&#x306E;&#x8868;&#x73FE;&#x306E;&#x7403;&#x306E;&#x9AD8;&#x30EC;&#x30D9;&#x30EB;&#x304B;&#x3089;&#x6D3E;&#x751F;&#x3057;&#x305F;&#x7D30;&#x80DE;&#x3092;&#x5206;&#x5316;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;CD44 &#x3068; CD133 &#x9AD8;&#x3044;&#x7403;&#x306E;&#x5F62;&#x6210;&#x306B;&#x8868;&#x73FE;&#x3055;&#x308C;&#x305F;&#x5206;&#x5316;&#x3057;&#x305F;&#x7D30;&#x80DE;&#x5BFE;&#x7D30;&#x80DE;&#x3002;&#x7403;&#x5F62;&#x6210;&#x7D30;&#x80DE;&#x3088;&#x308A;&#x9AD8;&#x3044; chemoresistance &#x3092;&#x7570;&#x306A;&#x308A;&#x30AB;&#x30EB;&#x30DC;&#x30D7;&#x30E9;&#x30C1;&#x30F3;&#x5206;&#x5316;&#x7D30;&#x80DE;&#x3068;&#x306F;&#x5BFE;&#x7167;&#x7684;&#x306B;&#x8868;&#x793A;&#x3002;&#x307E;&#x305F;&#x3001;&#x773C;&#x5185; NOD/SCID &#x30DE;&#x30A6;&#x30B9;&#x306B;&#x6CE8;&#x5165;&#x306E;&#x6570;&#x3068;&#x3057;&#x3066; 2 x 103 &#x7403;&#x5F62;&#x6210;&#x7D30;&#x80DE;&#x306F;&#x65B0;&#x3057;&#x3044;&#x816B;&#x760D;&#x3001;&#x5143;&#x60A3;&#x8005;&#x306E;&#x816B;&#x760D;&#x306B;&#x985E;&#x4F3C;&#x3059;&#x308B;&#x4E0A;&#x6607;&#x3092;&#x4E0E;&#x3048;&#x305F;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x7D50;&#x679C;&#x304B;&#x3089;&#x7403;&#x5F62;&#x6210;&#x7D30;&#x80DE;&#x306E;&#x5E79;&#x7D30;&#x80DE;&#x306E;&#x30D7;&#x30ED;&#x30D1;&#x30C6;&#x30A3;&#x3084;&#x816B;&#x760D;&#x5316;&#x3001;&#x9577;&#x671F;&#x57F9;&#x990A;&#x5F8C;&#x3082;&#x4FDD;&#x6301;&#x3059;&#x308B;&#x3053;&#x3068;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x3053;&#x308C;&#x306F;&#x7DB2;&#x819C;&#x82BD;&#x7D30;&#x80DE;&#x816B;&#x306B;&#x304A;&#x3051;&#x308B;&#x764C;&#x5E79;&#x7D30;&#x80DE;&#x306E;&#x7814;&#x7A76;&#x3001;&#x5316;&#x5B66;&#x7642;&#x6CD5;&#x85AC;&#x3068;&#x6226;&#x7565;&#x3092;&#x958B;&#x767A;&#x3059;&#x308B;&#x306E;&#x306B;&#x9069;&#x3057;&#x305F; in vitro &#x30E2;&#x30C7;&#x30EB;&#x306B;&#x306A;&#x308A;&#x307E;&#x3059;&#x3002;

&#x7DB2;&#x819C;&#x306E;&#x764C;&#x5E79;&#x7D30;&#x80DE;&#x306E;&#x3088;&#x3046;&#x306A;&#x30D7;&#x30ED;&#x30D1;&#x30C6;&#x30A3;&#x3092;&#x4ECB;&#x3057;&#x3066;&#x9577;&#x671F;&#x306E;&#x7121;&#x8840;&#x6E05;&#x57F9;&#x990A;&#x3072;&#x3068;&#x7DB2;&#x819C;&#x82BD;&#x7D30;&#x80DE;&#x816B;&#x304B;&#x3089;&#x306E;&#x30E1;&#x30F3;&#x30C6;&#x30CA;&#x30F3;&#x30B9;&#x3002;

&#xC774;&#xC804; &#xD559;&#xBB38;&#xC740; &#xC554; &#xC904;&#xAE30; &#xC138;&#xD3EC;&#xC640; &#xAC19;&#xC740; &#xC138;&#xD3EC;&#xC758; &#xC791;&#xC740; &#xC778;&#xAD6C; retinoblastoma&#xC5D0; &#xC788;&#xB294; &#xC124;&#xBA85; &#xD588;&#xB2E4;. &#xC774; &#xC778;&#xAD6C; &#xACF5;&#xBD80;&#xB97C; &#xC704;&#xD55C; &#xBAA8;&#xB378;&#xC744; &#xC81C;&#xACF5; &#xD558; &#xC6B0;&#xB9AC;&#xC5D0;&#xC11C; &#xC554; &#xC904;&#xAE30; &#xC138;&#xD3EC;&#xC640; &#xAC19;&#xC740; &#xC18D;&#xC131;&#xC744; &#xAC00;&#xC9C4; &#xC778;&#xAC04;&#xC758; retinoblastoma &#xC7A5;&#xAE30; &#xBB38;&#xD654;&#xB97C; &#xD655;&#xB9BD; &#xD558;&#xACE0;&#xC790; &#xD588;&#xB2E4;. &#xC2E0;&#xC120;&#xD55C; &#xC885;&#xC591; &#xC870;&#xC9C1; &#xC18C;&#xD654; &#xB418;&#xC5C8;&#xACE0; &#xD608; &#xCCAD; &#xBB34;&#xB8CC; &#xB9E4;&#xCCB4;&#xC5D0;&#xC11C; &#xAD50;&#xC591;. &#xC885;&#xC591; &#xBD84;&#xC57C; &#xD615;&#xC131; &#xD558; &#xACE0; &#xC9C0;&#xC18D;&#xC801;&#xC73C;&#xB85C; passaged &#xD588;&#xB2E4;. &#xC904;&#xAE30; &#xC138;&#xD3EC; &#xC18D;&#xC131;&#xC744; immunostaining, &#xC2E4;&#xC2DC;&#xAC04; &#xC815;&#xB7C9; RT-PCR &#xBC0F; chemoresistance &#xBD84;&#xC11D; &#xACB0;&#xACFC; &#xD1B5;&#xD574; &#xC870;&#xC0AC; &#xD588;&#xB2E4;. &#xC885;&#xC591; &#xBD84;&#xC57C;&#xB97C; &#xD615;&#xC131; &#xC138;&#xD3EC;&#xC758; tumorigenicity &#xB1CC;&#xC885;&#xC591; &#xC2E4;&#xD5D8;&#xC5D0; &#xC758;&#xD574; &#xD655;&#xC778; &#xB418;&#xC5C8;&#xB2E4;. &#xB610;&#xD55C;, &#xC6B0;&#xB9AC;&#xB294; &#xC138;&#xD3EC; &#xD45C;&#xBA74; &#xB9C8;&#xCEE4; CD44 &#xBC0F; CD133 &#xC2DD; &#xAC80;&#xC0AC;. &#xC885;&#xC591; &#xC138;&#xD3EC;&#xB294; 30 &#xAC1C; &#xC774;&#xC0C1;&#xC758; &#xAD6C;&#xC808;&#xC5D0; &#xB300; &#xD55C; &#xAD6C;&#xCCB4;&#xB97C; &#xBD80;&#xB3D9;&#xC73C;&#xB85C; &#xD655;&#xC7A5;. &#xC140; &#xC601;&#xC5ED;&#xC744; &#xD615;&#xC131; overexpressed &#xC904;&#xAE30; &#xC138;&#xD3EC; &#xC720;&#xC804;&#xC790; Oct&#x2011;4, Nestin &#xBC0F; Pax6. &#xBD84;&#xC57C;&#xC758; Immunostaining Nestin, Pax6 &#xBC0F; abcg2&#xC5D0; &#xB300; &#xD55C; &#xC591;&#xC131;&#xC744; &#xBCF4;&#xC600;&#xB2E4;. &#xBC18;&#xBA74;, &#xC774;&#xB7EC;&#xD55C; &#xD45C;&#xD604; &#xBD84;&#xC57C; &#xC0C1;&#xBD80; &#xC131;&#xC219;&#xD55C; &#xB9DD;&#xB9C9; &#xC138;&#xD3EC; &#xB9C8;&#xCEE4; MAP2, GFAP, recoverin, Opsin B &#xBC0F; &#xAD6D;&#xAC00; &#xC9C0;&#xC815; &#xC5F0;&#xAD6C;&#xC2E4;, &#xADF8;&#xB9AC;&#xACE0; NF200 GFAP, recoverin&#xC5D0; &#xB300; &#xD55C; &#xBCF4;&#xC600;&#xB2E4; immunoreactivity &#xBC0F; PKC&alpha;&#xC5D0;&#xC11C;&#xC5D0;&#xC11C; &#xD30C;&#xC0DD; &#xB41C; &#xC140; &#xAD6C;&#xBD84; &#xD569;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, CD44 &#xBC0F; CD133 &#xB192;&#xC740; &#xAD6C;&#xD615;&#xC744; &#xD615;&#xC131; &#xD45C;&#xD604; &#xD588;&#xB2E4; &#xCC28;&#xBCC4;&#xD654; &#xB41C; &#xC140; &#xB300; &#xC140;. &#xAD6C; &#xD615;&#xC131; &#xC138;&#xD3EC; &#xCC28;&#xBCC4;&#xD654; &#xB41C; &#xC140; &#xBC18;&#xB300;&#xB85C; carboplatin &#xB192;&#xC740; chemoresistance &#xD45C;&#xC2DC;. &#xB610;&#xD55C;, &#xB044; &#xB355; &#xC784;/SCID &#xC950;&#xB85C; &#xC801;&#xC740; 2 x 103 &#xAD6C; &#xD615;&#xC131; &#xC138;&#xD3EC;&#xC758; &#xC548; &#xAD6C;&#xB0B4; &#xC8FC;&#xC0AC; &#xC0C8;&#xB85C;&#xC6B4; &#xC885;&#xC591; &#xBE44;&#xC2B7;&#xD569;&#xB2C8;&#xB2E4;-&#xC6D0;&#xB798; &#xD658;&#xC790; &#xC885;&#xC591; &#xC0C1;&#xC2B9;&#xC744; &#xD588;&#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xACB0;&#xACFC; &#xAD6C;&#xCCB4;&#xB97C; &#xD615;&#xC131; &#xC138;&#xD3EC; &#xC904;&#xAE30; &#xC138;&#xD3EC; &#xC18D;&#xC131; &#xBC0F; tumorigenicity, &#xC7A5;&#xAE30; &#xBB38;&#xD654; &#xD6C4;&#xC5D0; &#xBCF4;&#xC874; &#xBC1D;&#xD614;&#xB2E4;. &#xC774; &#xC554; &#xC904;&#xAE30; &#xAC19;&#xC740; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; retinoblastoma &#xACF5;&#xBD80; &#xD558; &#xACE0; &#xD654;&#xD559;&#xC694;&#xBC95; &#xC57D;&#xBB3C; &#xBC0F; &#xC804;&#xB7B5; &#xAC1C;&#xBC1C; &#xC801;&#xD569; &#xD55C; &#xCCB4; &#xC678; &#xBAA8;&#xB378; &#xAC83;&#xC785;&#xB2C8;&#xB2E4;.

&#xC778;&#xAC04;&#xC758; retinoblastoma&#xC5D0;&#xC11C; &#xC7A5;&#xAE30; &#xD608; &#xCCAD; &#xBB34;&#xB8CC; &#xBB38;&#xD654;&#xB97C; &#xD1B5;&#xD574; &#xB9DD;&#xB9C9; &#xC554; &#xC904;&#xAE30; &#xC138;&#xD3EC;&#xC640; &#xAC19;&#xC740; &#xC18D;&#xC131;&#xC758; &#xC720;&#xC9C0; &#xBCF4;&#xC218;.

Estudios anteriores han demostrado que existe una poblaci&oacute;n peque&ntilde;a de c&eacute;lulas de c&aacute;ncer similares a c&eacute;lulas madre en retinoblastoma. Para proporcionar un modelo para el estudio de esta poblaci&oacute;n, se intent&oacute; establecer una cultura a largo plazo de retinoblastoma humana que tienen propiedades similares a c&eacute;lulas madre de c&aacute;ncer. Tejido tumoral fresco fue digerido y cultivada en medio sin suero. Esferas de tumor forman y fueron pasadas continuamente. Propiedades de c&eacute;lulas madre fueron examinados a trav&eacute;s de inmunotinci&oacute;n, RT-PCR cuantitativa en tiempo real y an&aacute;lisis quimiorresistencia. Tumorigenicidad de las c&eacute;lulas formadoras de esfera de tumor fue confirmada por experimentos de xenoinjerto. Adem&aacute;s, examinamos la expresi&oacute;n de marcadores de superficie celular CD44 y CD133. Las c&eacute;lulas tumorales ampliaron como flotante esferas para m&aacute;s de 30 pasajes. C&eacute;lulas formadoras de esfera sobreexpresa genes de la c&eacute;lula de v&aacute;stago Oct&#x2011;4, Nestin y Pax6. Inmunotinci&oacute;n de esferas positividad para Nestin, Pax6 y tambi&eacute;n ABCG2. En cambio, se distinguen las c&eacute;lulas derivadas de estos esferas expresadas altos niveles de los marcadores de c&eacute;lulas retinianas maduro MAP2, GFAP, recoverin, B opsina y Nrl y mostr&oacute; inmunoreactividad para NF200, GFAP, recoverin y PKC&alpha;. Adem&aacute;s, CD44 tanto CD133 altamente se expresaron en forma de esfera c&eacute;lulas vs c&eacute;lulas diferenciadas. C&eacute;lulas formadoras de esfera muestran mayor quimiorresistencia al carboplatino en comparaci&oacute;n con las c&eacute;lulas diferenciadas. Por otra parte, la inyecci&oacute;n intraocular de tan s&oacute;lo 2 x 103 c&eacute;lulas esfera en ratones NOD/SCID dio lugar a nuevos tumores similares a los tumores de pacientes originales. Estos resultados revelaron que las c&eacute;lulas formadoras de esfera conserven sus propiedades de c&eacute;lulas madre y la colonizaci&oacute;n, incluso despu&eacute;s de la cultura a largo plazo. Esto ser&iacute;a un modelo in vitro adecuado para estudiar las c&eacute;lulas de v&aacute;stago-como c&aacute;ncer en retinoblastoma y desarrollar estrategias y drogas quimioterap&eacute;uticas.

Mantenimiento de propiedades similares a c&eacute;lulas madre de c&aacute;ncer de retina a trav&eacute;s de cultura libre de suero a largo plazo de retinoblastoma humana.

This study deduced the pathological and immunophenotypic features of idiopathic dacryoadenitis, and researched into the possible immune mechanism of this lesion.

Histopathology and immunohistochemical profile in idiopathic dacryoadenitis.

To investigate the treatment status and prognosis of space-occupying lacrimal gland lesions at one tertiary eye center in China.

Space-occupying lesions of the lacrimal gland at one tertiary eye center in China: a retrospective clinical study of 95 patients.

Orbital solitary fibrous tumor (SFT) is a rare tumor and may recur or undergo malignant transformation without complete excision. We present a case of orbital SFT which recurred twice and underwent malignant transformation. The patient was treated with en bloc excision via a lateral orbitotomy. The postoperative histopathologic diagnosis of this case was an adult fibrosarcoma. Postoperative adjuvant radiation therapy was given. In 18 months of further follow-up, there has been no evidence of recurrence, both clinically and in regular imaging studies.

Malignant transformation of orbital solitary fibrous tumor.

To study the change of tear film and lacrimal secretion after lacrimal gland tumor removal operation.

[Tear film and lacrimal excretion changes after lacrimal gland tumor removal].

To evaluate the safety and efficacy of intralesional pingyangmycin for treatment of orbital vascular malformations.

Treatment of orbital vascular malformations with intralesional injection of pingyangmycin.

Radiation therapy is the most widely used and effective treatment for orbital tumors, but it causes dry eye due to lacrimal gland damage. Induced pluripotent stem cell-derived conditioned medium (iPSC-CM) has been shown to rescue different types of tissue damage. The present study investigated the mechanism of the potential radioprotective effect of IPS cell-derived conditioned medium (iPSC-CM) on gamma-irradiation-induced lacrimal gland injury (RILI) in experimental mice. In this study, we found that iPSC-CM ameliorated RILI. iPSC-CM markedly decreased radiotherapy induced inflammatory processes, predominantly through suppressing p38/JNK signaling. Further signaling pathway analyses indicated that iPSC-CM could suppress Akt (Protein Kinase B, PKB) phosphorylation. High levels of midkine (MDK) were also found in iPSC-CM and could be involved in lacrimal gland regeneration by promoting cell migration and proliferation. Thus, our study indicates that inhibiting the p38/JNK pathway or increasing the MDK level might be a therapeutic target for radiation-induced lacrimal gland injury.

Improvement of radiotherapy-induced lacrimal gland injury by induced pluripotent stem cell-derived conditioned medium via MDK and inhibition of the p38/JNK pathway.

To establish a radiation-injured model of lacrimal gland in mice and explore its pathophysiology.

[Establishment mice model of radiation-injured lacrimal gland and pathophysiological exploration].

To evaluate the tear-film meniscus with optical coherence tomography (OCT) in patients with idiopathic dacryoadenitis and to determine its relationships with the clinical tests.

Changes in Tear Film Characteristics in Patients With Idiopathic Dacryoadenitis.

Retinoblastoma protein (Rb) is a prototypical tumor suppressor that is vital to the negative regulation of the cell cycle and tumor progression. Hypo-phosphorylated Rb is associated with G0/G1 arrest by suppressing E2F transcription factor activity, whereas Rb hyper-phosphorylation allows E2F release and cell cycle progression from G0/G1 to S phase. However, the factors that regulate cyclin-dependent protein kinase (CDK)-dependent hyper-phosphorylation of Rb during the cell cycle remain obscure. In this study, we show that throughout the cell cycle, Rb is specifically small ubiquitin-like modifier (SUMO)ylated at early G1 phase. SUMOylation of Rb stimulates its phosphorylation level by recruiting a SUMO-interaction motif (SIM)-containing kinase CDK2, leading to Rb hyper-phosphorylation and E2F-1 release. In contrast, a SUMO-deficient Rb mutant results in reduced SUMOylation and phosphorylation, weakened CDK2 binding, and attenuated E2F-1 sequestration. Furthermore, we reveal that Rb SUMOylation is required for cell proliferation. Therefore, our study describes a novel mechanism that regulates Rb phosphorylation during cell cycle progression.

SUMOylation of Rb enhances its binding with CDK2 and phosphorylation at early G1 phase.

To investigate the application of Rose and Wright's algorithm in diagnosing lacrimal gland masses.

Article	Year
Expression of interleukin-17 in autoimmune dacryoadenitis in MRL/lpr mice. Current eye research\| PubMed ID: 20858106	2010
Maintenance of retinal cancer stem cell-like properties through long-term serum-free culture from human retinoblastoma. Oncology reports\| PubMed ID: 21573498	2011
Histopathology and immunohistochemical profile in idiopathic dacryoadenitis. Current eye research\| PubMed ID: 22510006	2012
Space-occupying lesions of the lacrimal gland at one tertiary eye center in China: a retrospective clinical study of 95 patients. International journal of ophthalmology\| PubMed ID: 22762051	2012
Malignant transformation of orbital solitary fibrous tumor. International ophthalmology\| PubMed ID: 23525957	2013
[Tear film and lacrimal excretion changes after lacrimal gland tumor removal]. [Zhonghua yan ke za zhi] Chinese journal of ophthalmology\| PubMed ID: 23601462	2013
Treatment of orbital vascular malformations with intralesional injection of pingyangmycin. The British journal of ophthalmology\| PubMed ID: 23603483	2013
Improvement of radiotherapy-induced lacrimal gland injury by induced pluripotent stem cell-derived conditioned medium via MDK and inhibition of the p38/JNK pathway. International journal of molecular sciences\| PubMed ID: 25314301	2014
[Establishment mice model of radiation-injured lacrimal gland and pathophysiological exploration]. [Zhonghua yan ke za zhi] Chinese journal of ophthalmology\| PubMed ID: 25547579	2014
Changes in Tear Film Characteristics in Patients With Idiopathic Dacryoadenitis. Ophthalmic plastic and reconstructive surgery\| PubMed ID: 26863039	2017
SUMOylation of Rb enhances its binding with CDK2 and phosphorylation at early G1 phase. Cell cycle (Georgetown, Tex.)\| PubMed ID: 27163259	2016
Application of Rose and Wright's algorithm in the diagnosis of lacrimal gland masses: a study of 93 cases. Canadian journal of ophthalmology. Journal canadien d'ophtalmologie\| PubMed ID: 28237145	2017

Jiang Qian

Biography

JoVE Journal Publications

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