cytotoxic-efficacy-photodynamic-therapy-osteosarcoma-cells

Cytotoxic Efficacy of Photodynamic Therapy in Osteosarcoma Cells <em>In Vitro</em>

In recent years, there has been the difficulty in finding more effective therapies against cancer with less systemic side effects. Therefore Photodynamic ...

Balgrist University Hospital, Zurich, Switzerland

&#x5149;&#x52A8;&#x529B;&#x7597;&#x6CD5; (PDT) &#x662F;&#x6838;&#x51C6;&#x59D1;&#x606F;&#x6027;&#x548C;&#x6CBB;&#x7597;&#x6027;&#x6CBB;&#x7597;&#x7684;&#x67D0;&#x4E9B;&#x5F62;&#x5F0F;&#x7684;&#x5F53;&#x5730;&#x764C;&#x75C7;&#x3001; &#x764C;&#x524D;&#x75C5;&#x53D8;&#x548C; nononcological &#x75BE;&#x75C5;&#x5FAE;&#x521B;&#x6CBB;&#x7597;&#x65B9;&#x6CD5;&#x3002;&#x4F5C;&#x4E3A;&#x4ECA;&#x540E;&#x7684;&#x7814;&#x7A76;&#x4E2D;&#x9488;&#x5BF9; PDT &#x9AA8;&#x8089; (OS)&#xFF0C;&#x5728;&#x672C;&#x7814;&#x7A76;&#x62A5;&#x544A;&#x5728;&#x672F;&#x4E2D;&#x5E94;&#x7528;&#x52A8;&#x7269;&#x6A21;&#x578B;&#x7684;&#x5148;&#x51B3;&#x6761;&#x4EF6;&#xFF0C;&#x6211;&#x4EEC;&#x8C03;&#x67E5;&#x4E86;&#x5438;&#x6536;&#x548C;&#x9ED1;&#x6697;&#x548C;&#x7167;&#x7247;-&#x6BD2;&#x6027;&#x7684;&#x5149;&#x654F;&#x5242; mTHPC &#x5728;&#x8F6C;&#x79FB;&#x6027;&#x4EBA;&#x7C7B; OS &#x7684;&#x5355;&#x5143;&#x683C;&#x884C; 143B&#xFF0C;&#x8F6C;&#x8F7D;&#xFF0C;intratibially &#x6CE8;&#x5165; SCID &#x5C0F;&#x9F20;&#x81EA;&#x53D1;&#x6027;&#x3001; &#x4FB5;&#x7565;&#x6027;&#x80BA;&#x8F6C;&#x79FB;&#xFF0C;OS &#x60A3;&#x8005;&#x6B7B;&#x4EA1;&#x7684;&#x4E3B;&#x8981;&#x539F;&#x56E0;&#x3002;MTHPC 143B &#x7EC6;&#x80DE;&#x5438;&#x6536;&#x4E86;&#x65F6;&#x95F4;&#x548C;&#x5242;&#x91CF;&#x4F9D;&#x8D56;&#x6027;&#x3002;mTHPC &#x79EF;&#x7D2F;&#x5230; 143B &#x6BD4;&#x7236;&#x6BCD;&#x4F4E;&#x8F6C;&#x79FB;&#x5C45;&#x764C;&#x7EC6;&#x80DE;&#x7684;&#x8F83;&#x9AD8;&#x6C34;&#x5E73;&#x3002;143B &#x7EC6;&#x80DE;&#xFF0C;&#x53CD;&#x6620; mTHPC &#x9ED1;&#x6697;&#x7684;&#x6BD2;&#x6027;&#xFF0C;&#x5927;&#x5927;&#x51CF;&#x5C11;&#x53D1;&#x751F;&#x540E;&#x5728; mTHPC &#x6D53;&#x5EA6; &ge;2.5 &mu; mL(-1) &#x5728;&#x9ED1;&#x6697;&#x4E2D;&#x5B75;&#x5316;&#x3002;&#x5149;&#x6BD2;&#x6027;&#x5B9E;&#x9A8C;&#x4E0E;&#x7167;&#x660E;&#x7684; 652 nm &#x6FC0;&#x5149;&#x706F; (2.5-10 J cm(-2))&#x3001; &#x534A;-&#x6781;&#x5927;&#x81F4;&#x547D;&#x5242;&#x91CF;&#x7684;&#x4ECB;&#x4E4E; 0.012 0.047 &mu; mL(-1) &#x81F3; mTHPC&#x3002;&#x8FD9;&#x79CD;&#x6CBB;&#x7597;&#x6FC0;&#x6D3B;&#x534A;&#x80F1;&#x6C28;&#x9178;&#x86CB;&#x767D;&#x9176;-3&#xFF0C;-7-9 &#x548C; Z VAD-&#x82AC;&#x5170;&#x9A6C;&#x514B; inhibitable &#x6CE2;&#x5170;&#x7684;&#x4E73;&#x6C9F;&#x91CC;&#xFF0C;&#x6307;&#x793A;&#x4F9D;&#x8D56;&#x534A;&#x80F1;&#x6C28;&#x9178;&#x86CB;&#x767D;&#x9176;&#x7684;&#x7EC6;&#x80DE;&#x51CB;&#x4EA1;&#x3002;&#x603B;&#x4E4B;&#xFF0C;&#x4E0E; mTHPC &#x5149;&#x52A8;&#x529B;&#x7597;&#x6CD5;&#x53EF;&#x4EE5;&#x6709;&#x6548;&#x5730;&#x8F6C;&#x79FB; 143B &#x4EBA;&#x6210;&#x9AA8;&#x8089;&#x7624;&#x7EC6;&#x80DE;&#x7CFB;&#x4F53;&#x5916;&#x3002;

&#x5728;&#x8F6C;&#x79FB;&#x6027;&#x4EBA;&#x7C7B; 143B &#x9AA8;&#x8089;&#x7624;&#x7EC6;&#x80DE; mTHPC &#x4ECB;&#x5BFC;&#x7684;&#x5149;&#x52A8;&#x529B;&#x7597;&#x6CD5;&#x662F;&#x6709;&#x6548;&#x3002;

Photodynamic therapy (PDT) is a minimally invasive therapeutic modality approved for palliative and curative treatment of some forms of local cancers, precancerous lesions and nononcological disorders. As a prerequisite for future studies in animal models aiming at an intraoperative application of PDT in osteosarcoma (OS), in the present study, we investigated the uptake and the dark- and photo-toxicity of the photosensitizer mTHPC in the metastatic human OS cell line 143B, which, intratibially injected into SCID mice, reproduces spontaneous, aggressive lung metastasis, the main cause of death in OS patients. The uptake of mTHPC by 143B cells was time- and dose-dependent. mTHPC accumulated to higher levels in the 143B than in the parental low-metastatic HOS cell line. A significant decrease in viability of 143B cells, reflecting mTHPC dark-toxicity, occurred upon incubation in the dark at mTHPC concentrations â‰¥2.5 Î¼g mL(-1). In phototoxicity experiments with illumination by 652 nm laser light (2.5-10 J cm(-2)), the half-maximal lethal doses of mTHPC ranged from 0.012 to 0.047 Î¼g mL(-1). This treatment activated caspase-3, -7 and -9 and Z-VAD-FMK-inhibitable PARP cleavage, indicating caspase-dependent apoptosis. In conclusion, PDT with mTHPC is effective in the metastatic 143B human osteosarcoma cell line in vitro.

mTHPC-mediated photodynamic therapy is effective in the metastatic human 143B osteosarcoma cells.

La th&eacute;rapie photodynamique (PDT) est une modalit&eacute; th&eacute;rapeutique mini-invasive, approuv&eacute;e pour le traitement palliatif et curatif de certaines formes de cancers les, les l&eacute;sions pr&eacute;canc&eacute;reuses et les troubles nononcological. Comme condition pr&eacute;alable pour de futures &eacute;tudes sur des mod&egrave;les animaux qui vise &agrave; une application perop&eacute;ratoire de PDT dans l&#39;ost&eacute;osarcome (OS), dans la pr&eacute;sente &eacute;tude, nous avons &eacute;tudi&eacute; l&#39;absorption et la noir-photo-toxicit&eacute; et de la mTHPC photosensibilisateur dans la lign&eacute;e de cellules de OS humaine m&eacute;tastatique 143 ter, qui, intratibially inject&eacute;e &agrave; des souris SCID, reproduit les m&eacute;tastases pulmonaires spontan&eacute;e, agressif, la principale cause de d&eacute;c&egrave;s chez les patients de l&#39;OS. La captation de mTHPC par les cellules de 143 ter est et dose-d&eacute;pendante du temps. mTHPC accumul&eacute;s &agrave; des niveaux plus &eacute;lev&eacute;s dans les 143 ter que dans les HOS m&eacute;tastatique faible parental lign&eacute;e cellulaire. Une diminution significative de la viabilit&eacute; des cellules de 143 ter, qui refl&egrave;te mTHPC dark-toxicit&eacute;, s&#39;est produite lors de l&#39;incubation &agrave; l&#39;obscurit&eacute; &agrave; mTHPC les concentrations &ge;2 &mu;g mL(-1). Dans les exp&eacute;riences de phototoxicit&eacute; avec &eacute;clairage par la lumi&egrave;re laser de 652 nm (2,5 &agrave; 10 J cm(-2)), les mi-hauteur des doses l&eacute;tales de mTHPC variaient entre 0,012 et 0,047 &mu;g mL(-1). Ce traitement activ&eacute; caspase-3, -7 et -9 et le clivage inhibitable Z-VAD-marks finlandais par PARP, indiquant l&#39;apoptose d&eacute;pendante de la caspase. En conclusion, les PDT avec mTHPC est efficace dans la lign&eacute;e de cellules d&#39;ost&eacute;osarcome humain m&eacute;tastatique 143 ter in vitro.

mTHPC par l&#39;interm&eacute;diaire de la th&eacute;rapie photodynamique est efficace dans les cellules d&#39;ost&eacute;osarcome m&eacute;tastatique 143 humaine b.

Photodynamische Therapie (PDT) ist eine minimal-invasive therapeutische Modalit&auml;t f&uuml;r palliative und kurative Behandlung bestimmter Formen von lokalen Krebsarten, pr&auml;kanzer&ouml;sen L&auml;sionen und Nononcological Erkrankungen zugelassen. Als Voraussetzung f&uuml;r zuk&uuml;nftige Studien an Tiermodellen abzielen, eine intraoperative Anwendung von PDT im Osteosarkom (OS), in der vorliegenden Studie untersuchten wir Verbreitung und die Dunkelheit - und Foto-Toxizit&auml;t der Photosensibilisator-mTHPC in die metastasierendem menschlichen OS Zelllinie 143 b, die Intratibially eingespritzt SCID-M&auml;use, spontan, aggressiven Lungenkrebs Metastasen, die h&auml;ufigste Todesursache bei OS Patienten reproduziert. Die Aufnahme von mTHPC von 143 b-Zellen war Zeit und Dosis-abh&auml;ngige. mTHPC angesammelt auf h&ouml;heren Ebenen der 143 b als in der elterlichen niedrig-metastasiertem HOS Zeile Zelle. Ein deutlichen R&uuml;ckgang der Lebensf&auml;higkeit von 143 b-Zellen, reflektieren mTHPC dunkel-Toxizit&auml;t, trat nach Inkubation im Dunkeln bei mTHPC Konzentrationen &ge;2.5 &mu;g mL(-1). Phototoxizit&auml;t Experimente mit Beleuchtung von 652 nm Laserlicht (2.5-10-J-cm(-2)), die halb-maximale letalen Dosen von mTHPC reichten von 0,012 bis 0,047 &mu;g mL(-1). Diese Behandlung aktiviert Caspase-3,-7 und-9 und Z-VAD-FMK-inhibitable PARP-Spaltung, Angabe Caspase-abh&auml;ngigen Apoptose. Abschlie&szlig;end ist die PDT mit mTHPC effektiv in die metastasierendem 143 menschlichen Osteosarkom Zelllinie in-vitro.

mTHPC-vermittelte photodynamische Therapie ist in den metastasierendem menschlichen 143 Osteosarkom Zellen wirksam.

Terapia fotodinamica (PDT) &egrave; una modalit&agrave; terapeutica mininvasiva approvata per il trattamento palliativo e curativo di alcune forme di tumori locali, lesioni precancerose e disturbi nononcological. Come prerequisito per futuri studi in animali modelli volti a un&#39;applicazione intraoperatoria di PDT in osteosarcoma (OS), nel presente studio, abbiamo studiato l&#39;assorbimento e la dark-foto-tossicit&agrave; e della mTHPC fotosensibilizzatore in OS metastatico umano cella linea 143B, che, intratibially iniettato in topi SCID, riproduce spontanea, metastasi polmonare aggressiva, la principale causa di morte nei pazienti di OS. L&#39;assorbimento di mTHPC dalle cellule 143B era tempo - e dose-dipendente. mTHPC accumulato a livelli superiori nella 143B rispetto nel parentale HOS basso-metastatico cella linea. Una diminuzione significativa nell&#39;attuabilit&agrave; delle cellule 143B, riflettendo mTHPC dark-tossicit&agrave;, si &egrave; verificato dopo incubazione al buio a mTHPC concentrazioni &ge;2.5 &mu;g mL(-1). In esperimenti di fototossicit&agrave; con illuminazione da 652 luce laser nm (2,5-10 J cm(-2)), la met&agrave;-maximal dosi letali di mTHPC variava da 0,012 a 0,047 &mu;g mL(-1). Questo trattamento attivato caspase-3, -7 e -9 e Z-VAD-FMK-inhibitable PARP scissione, indicando l&#39;apoptosi caspasi-dipendente. In conclusione, PDT con mTHPC &egrave; efficace per la linea di cellule osteosarcoma umano 143B metastatico in vitro.

terapia fotodinamica mTHPC-mediata &egrave; efficacia in cellule di osteosarcoma metastatico umano 143B.

&#x5149;&#x7DDA;&#x529B;&#x5B66;&#x7642;&#x6CD5; &#xFF08;PDT&#xFF09; &#x306F;&#x4F4E;&#x4FB5;&#x8972;&#x6CBB;&#x7642;&#x30E2;&#x30C0;&#x30EA;&#x30C6;&#x30A3; &#x30ED;&#x30FC;&#x30AB;&#x30EB;&#x764C;&#x3001;&#x524D;&#x764C;&#x75C5;&#x5909;&#x3068; nononcological &#x969C;&#x5BB3;&#x306E;&#x3044;&#x304F;&#x3064;&#x304B;&#x306E;&#x5F62;&#x614B;&#x306E;&#x7DE9;&#x548C;&#x3068;&#x6CBB;&#x7652;&#x306E;&#x6CBB;&#x7642;&#x306E;&#x627F;&#x8A8D;&#x3067;&#x3059;&#x3002;&#x5C06;&#x6765;&#x7814;&#x7A76;&#x52D5;&#x7269;&#x30E2;&#x30C7;&#x30EB;&#x306E;&#x73FE;&#x5728;&#x306E;&#x7814;&#x7A76;&#x3067;&#x306F;, PDT &#x306E;&#x8853;&#x4E2D;&#x30A2;&#x30D7;&#x30EA;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x9AA8;&#x8089;&#x816B; (OS) &#x3092;&#x76EE;&#x6307;&#x3057;&#x3066;&#x6211;&#x3005; &#x53D6;&#x308A;&#x8FBC;&#x307F;&#x3092;&#x8ABF;&#x3079;&#x305F;&#x3057;&#x3001;&#x6697;&#x3044;&#x6BD2;&#x6027;&#x3068;&#x5199;&#x771F;&#x306E;&#x5149;&#x5897;&#x611F;&#x5264; mTHPC &#x8EE2;&#x79FB;&#x6027;&#x306E;&#x4EBA;&#x9593;&#x306E; OS &#x3067;&#x306F;&#x3001;intratibially &#x3092; SCID &#x30DE;&#x30A6;&#x30B9;&#x306B;&#x6CE8;&#x5165;&#x3055;&#x308C;&#x305F;&#x81EA;&#x767A;&#x7684;&#x306A;&#x518D;&#x73FE;&#x30E9;&#x30A4;&#x30F3; 143B &#x7D30;&#x80DE;, &#x7A4D;&#x6975;&#x7684;&#x306A;&#x80BA;&#x8EE2;&#x79FB;, OS &#x60A3;&#x8005;&#x6B7B;&#x4EA1;&#x306E;&#x4E3B;&#x306A;&#x539F;&#x56E0;&#x306E;&#x305F;&#x3081;&#x306E;&#x524D;&#x63D0;&#x6761;&#x4EF6;&#x3068;&#x3057;&#x3066;&#x3002;MTHPC 143B &#x7D30;&#x80DE;&#x3067;&#x306E;&#x53D6;&#x308A;&#x8FBC;&#x307F;&#x6642;&#x9593;&#x3068;&#x7528;&#x91CF;&#x4F9D;&#x5B58;&#x3060;&#x3063;&#x305F;&#x3002;mTHPC &#x306F;&#x3001;&#x4E0A;&#x4F4D;&#x30EC;&#x30D9;&#x30EB;&#x306E;&#x89AA;&#x4F4E;&#x8EE2;&#x79FB; HOS &#x7D30;&#x80DE;&#x3088;&#x308A;&#x3082; 143B &#x306B;&#x84C4;&#x7A4D;&#x3002;MTHPC &#x6FC3;&#x5EA6; &ge;2.5 &mu; g/ml mL(-1) &#x3067;&#x6697;&#x95C7;&#x306E;&#x4E2D;&#x3067;&#x30A4;&#x30F3;&#x30AD;&#x30E5;&#x30D9;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x6642;&#x3001;&#x6697;&#x3044;&#x6BD2;&#x6027; mTHPC &#x3092;&#x53CD;&#x6620;&#x3057;&#x3066;&#x3001;143B &#x7D30;&#x80DE;&#x306E;&#x751F;&#x5B58;&#x7387;&#x306E;&#x6709;&#x610F;&#x306A;&#x6E1B;&#x5C11;&#x304C;&#x767A;&#x751F;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;652 Nm &#x30EC;&#x30FC;&#x30B6;&#x5149; &#xFF08;2.5 10 J cm(-2))&#x3001;mTHPC &#x304B;&#x3089; 0.012 0.047 &mu; g/ml &#x306E; mL(-1) &#x306B;&#x9577;&#x5C04;&#x7A0B;&#x306E;&#x534A;&#x6700;&#x5927;&#x81F4;&#x6B7B;&#x91CF;&#x306B;&#x3088;&#x308B;&#x7167;&#x660E;&#x5149;&#x6BD2;&#x6027;&#x5B9F;&#x9A13;&#x3067;&#x3002;&#x3053;&#x306E;&#x6CBB;&#x7642;&#x6CD5;&#x306F;&#x30AB;&#x30B9;&#x30D1;&#x30FC;&#x30BC; 3&#x3001;-7&#x3001;-9 &#x304A;&#x3088;&#x3073;&#x30AB;&#x30B9;&#x30D1;&#x30FC;&#x30BC;&#x4F9D;&#x5B58;&#x6027;&#x30A2;&#x30DD;&#x30C8;&#x30FC;&#x30B7;&#x30B9;&#x3092;&#x793A;&#x3059; Z VAD FMK inhibitable PARP &#x80F8;&#x306E;&#x8C37;&#x9593;&#x3092;&#x6D3B;&#x6027;&#x5316;&#x3002;&#x7D50;&#x8AD6;&#x3068;&#x3057;&#x3066;&#x3001;mTHPC &#x3068; PDT &#x306F;&#x3001;&#x8EE2;&#x79FB;&#x6027; 143B &#x3072;&#x3068;&#x9AA8;&#x8089;&#x816B;&#x7D30;&#x80DE;&#x682A; in vitro &#x3067;&#x52B9;&#x679C;&#x7684;&#x3067;&#x3059;&#x3002;

&#x5149;&#x7DDA;&#x529B;&#x5B66;&#x7642;&#x6CD5;&#x306E; mTHPC &#x3092;&#x4ECB;&#x3057;&#x305F;&#x4EBA;&#x9593; 143B &#x306E;&#x8EE2;&#x79FB;&#x6027;&#x9AA8;&#x8089;&#x816B;&#x7D30;&#x80DE;&#x3067;&#x306F;&#x52B9;&#x679C;&#x7684;&#x3067;&#x3059;&#x3002;

Photodynamic &#xCE58;&#xB8CC; (PDT) &#xC9C0;&#xC5ED; &#xC554;, &#xC804; &#xC554; &#xBCD1; &#xBCC0; &#xBC0F; nononcological &#xC9C8;&#xD658;&#xC758; &#xC77C;&#xBD80; &#xD615;&#xD0DC;&#xC758; &#xC644;&#xD654; &#xBC0F; &#xCE58;&#xB8CC; &#xCE58;&#xB8CC;&#xC5D0; &#xB300; &#xD55C; &#xC2B9;&#xC778;&#xC744; &#xCD5C;&#xC18C; &#xCE68; &#xC2B5; &#xCE58;&#xB8CC; &#xC591;&#xC2DD; &#xC801;&#xC784; &#xC774;&#xB2E4;. &#xD604;&#xC7AC;&#xC758; &#xC5F0;&#xAD6C;&#xC5D0; &#xB2E4;&#xB9AC; (OS)&#xC5D0; PDT&#xC758; &#xC790;&#xAC00; &#xC751;&#xC6A9;&#xC744; &#xBAA9;&#xD45C;&#xB85C; &#xD558;&#xB294; &#xB3D9;&#xBB3C; &#xBAA8;&#xB378;&#xC5D0;&#xC11C; &#xBBF8;&#xB798; &#xC5F0;&#xAD6C;&#xB97C; &#xC704;&#xD55C; &#xC804;&#xC81C; &#xC870;&#xAC74;&#xC73C;&#xB85C; &#xC6B0;&#xB9AC;&#xAC00; &#xAE00;&#xADC0;&#xC640; &#xB2E4;&#xD06C;-&#xBC0F; &#xC0AC;&#xC9C4;-&#xC758; &#xB3C5;&#xC131; &#xC804;&#xC774;&#xC131; &#xC778;&#xAC04; OS &#xC140; &#xB77C;&#xC778; 143B, photosensitizer Mthpc&#xB294; SCID &#xC950;&#xC5D0; &#xC8FC;&#xC785; &#xD558;&#xB294; intratibially &#xC7AC;&#xD604; &#xD55C; &#xC790;&#xBC1C;&#xC801;, &#xC801;&#xADF9;&#xC801;&#xC778; &#xD3D0; &#xC804;&#xC774;, OS &#xD658;&#xC790;&#xC5D0;&#xC11C; &#xC0AC;&#xB9DD;&#xC758; &#xC8FC;&#xC694; &#xC6D0;&#xC778;&#xC774; &#xC870;&#xC0AC; &#xD588;&#xB2E4;. 143B &#xC138;&#xD3EC;&#xC5D0; &#xC758;&#xD574; Mthpc&#xC758; &#xAE00;&#xADC0;&#xB294; &#xC2DC;&#xAC04; &#xBCF5;&#xC6A9;&#xB7C9;&#xC5D0; &#xC885;&#xC18D;. mTHPC 143B &#xBD80;&#xBAA8;&#xC758; &#xB0AE;&#xC740; &#xC804;&#xC774;&#xC131; &#xACC4; &#xC9D1;&#xC5D0; &#xC120; &#xC140; &#xBCF4;&#xB2E4; &#xB192;&#xC740; &#xC218;&#xC900;&#xC73C;&#xB85C; &#xCD95;&#xC801;. MTHPC &#xB2E4;&#xD06C;-&#xB3C5;&#xC131;, &#xBC18;&#xC601; 143B &#xC138;&#xD3EC;&#xC758; &#xC0DD;&#xC874;&#xC5D0; &#xD070; &#xAC10;&#xC18C; mTHPC &#xB18D;&#xB3C4; &ge;2.5 &mu;g Ml(-1)&#xC5D0;&#xC11C; &#xC5B4;&#xB460; &#xC18D;&#xC5D0;&#xC11C; &#xBD80; &#xD654;&#xB97C; &#xB530;&#xB77C; &#xBC1C;&#xC0DD; &#xD588;&#xC2B5;&#xB2C8;&#xB2E4;. (2.5-10 J cm(-2)), mTHPC &mu;g mL(-1) 0.047 0.012&#xC5D0;&#xC11C; &#xC6D0;&#xAC70;&#xB9AC;&#xC758; &#xC808;&#xBC18; &#xCD5C;&#xB300;&#xD55C; &#xCE58;&#xBA85;&#xC801;&#xC778; &#xBCF5;&#xC6A9;&#xB7C9; 652 nm &#xB808;&#xC774;&#xC800; &#xBE5B;&#xC5D0; &#xC758;&#xD574; &#xC870;&#xBA85; phototoxicity &#xC2E4;&#xD5D8;&#xC5D0;&#xC11C;. &#xC774; &#xCE58;&#xB8CC;&#xB294; caspase 3-7-9 &#xBC0F; caspase &#xC885;&#xC18D; apoptosis&#xB97C; &#xB098;&#xD0C0;&#xB0B4;&#xB294; Z-VAD-&#xC8FC;-inhibitable PARP &#xBD84;&#xC5F4;&#xC744; &#xD65C;&#xC131;&#xD654;. &#xB05D;&#xC73C;&#xB85C;, Mthpc&#xC640; PDT &#xC804;&#xC774;&#xC131; 143B &#xC778;&#xAC04;&#xC758; &#xB2E4;&#xB9AC; &#xC140; &#xB77C;&#xC778; &#xCCB4; &#xC678;&#xC5D0; &#xD6A8;&#xACFC;&#xC801;&#xC785;&#xB2C8;&#xB2E4;.

mTHPC-&#xC911;&#xC7AC; photodynamic &#xCE58;&#xB8CC;&#xB294; &#xC804;&#xC774;&#xC131; &#xC778;&#xAC04; 143B &#xB2E4;&#xB9AC; &#xC140;&#xC5D0; &#xD6A8;&#xACFC;&#xC801;&#xC785;&#xB2C8;&#xB2E4;.

La terapia fotodin&aacute;mica (PDT) es una modalidad terap&eacute;utica m&iacute;nimamente invasiva aprobada para el tratamiento paliativo y curativo de algunas formas de c&aacute;ncer local, lesiones precancerosas y trastornos nononcological. Como requisito previo para futuros estudios en modelos animales con el objetivo de un uso intraoperativo del PDT en osteosarcoma (OS), en el presente estudio, se investigan la absorci&oacute;n y la oscuridad y Foto-toxicidad del fotosensibilizador mTHPC en la l&iacute;nea de c&eacute;lula humana metast&aacute;tica de las OS 143B, que, intratibially inyectado en ratones de SCID, reproduce la met&aacute;stasis del pulm&oacute;n espont&aacute;neo, agresivo, la principal causa de muerte en pacientes OS. La captaci&oacute;n de mTHPC por c&eacute;lulas 143B era dependiente del tiempo y dosis. mTHPC acumulado a niveles m&aacute;s altos en el 143B de c&eacute;lulas en el HOS metast&aacute;sico baja parental. Se ha producido una disminuci&oacute;n significativa en la viabilidad de las c&eacute;lulas 143B, reflejando el mTHPC oscuro-toxicidad, sobre incubaci&oacute;n en la oscuridad en mTHPC concentraciones &ge;2. 5 &micro;g ml. En experimentos de fototoxicidad con iluminaci&oacute;n por luz laser de 652 nm (2.5-10 J cm(-2)), las dosis letales de medio-m&aacute;ximo de mTHPC vari&oacute; de 0.012 a 0.047 &mu;g ml. Este tratamiento activa la caspasa-3, -7 y -9 y escote PARP Z-VAD-FMK-inhibitable, indicando la apoptosis caspasa-dependiente. En conclusi&oacute;n, PDT con mTHPC es eficaz en la l&iacute;nea de c&eacute;lulas de osteosarcoma humano metast&aacute;tico 143B in vitro.

mTHPC-mediada de la terapia fotodin&aacute;mica es eficaz en las c&eacute;lulas de osteosarcoma metast&aacute;tico 143B humana.

Formation of metastases in the lungs is the major cause of death in patients suffering from osteosarcoma (OS). Metastases at presentation and poor response to preoperative chemotherapy are strong predictors for poor patient outcome. The elucidation of molecular markers that promote metastasis formation and/or chemoresistance is therefore of importance. CD44 is a plasma membrane glycoprotein that binds to the extracellular matrix component hyaluronan (HA) and has been shown to be involved in metastasis formation in a variety of other tumors. Here we investigated the role of CD44 expression on OS tumor formation and metastasis. High CD44 expression, evaluated with a tissue microarray including samples from 53 OS patients and stained with a pan-CD44 antibody (Hermes3), showed a tendency (p < 0.08) to shortened overall survival. However, nonresponders and patients with lung metastases and high CD44 expression had significantly poorer prognosis than patients with low CD44 expression. Overexpression of the standard CD44 isoform (CD44s) and its HA-binding defective mutant R41A in osteoblastic SaOS-2 cells resulted in HA-independent higher migration rates and increased chemoresistance, partially dependent on HA. In an orthotopic mouse model of OS, overexpression of CD44s in SaOS-2 cells resulted in an HA-dependent increased primary tumor formation and increased numbers of micrometastases and macrometastases in the lungs. In conclusion, although CD44 failed to be an independent predictor for patient outcome in this limited cohort of OS patients, increased CD44 expression was associated with even worse survival in patients with chemoresistance and with lung metastases. CD44-associated chemoresistance was also observed in vitro, and increased formation of lung metastases was found in vivo in SCID mice.

CD44 enhances tumor formation and lung metastasis in experimental osteosarcoma and is an additional predictor for poor patient outcome.

Osteosarcoma is the most common malignant bone tumor in children and young adults. Since the introduction of chemotherapy, the 5-year survival rate of patients with non-metastatic osteosarcoma is ~70%. The main problems in osteosarcoma therapy are the occurrence of metastases, severe side-effects and chemoresistance. Antiproliferative and apoptotic effects of quercetin were shown in several types of cancers, including breast cancer and lung carcinoma.

Evaluation of quercetin as a potential drug in osteosarcoma treatment.

Osteosarcoma (OS) is the most frequent primary malignant bone cancer in children and adolescents with a high propensity for lung metastasis. Therefore, it is of great importance to identify molecular markers leading to increased metastatic potential in order to devise more effective therapeutic strategies that suppress metastasis, the major cause of death in OS. CD44, the principal receptor for the extracellular matrix component hyaluronan (HA), is frequently found overexpressed in tumor cells and has been implicated in metastatic spread in various cancer types. Here, we investigated the effects of stable shRNA-mediated silencing of CD44 gene products on in vitro and in vivo metastatic properties of the highly metastatic human 143-B OS cell line. In vitro, CD44 knockdown resulted in a 73% decrease in the adhesion to HA, a 57% decrease in the migration rate in a trans-filter migration assay, and a 28% decrease in the cells' capacity for anchorage-independent growth in soft agar compared to the control cells, implicating that CD44 expression contributes to the metastatic activity of 143-B cells. However, making use of an orthotopic xenograft OS mouse model, we demonstrated that reduced CD44 expression facilitated primary tumor growth and formation of pulmonary metastases. The enhanced malignant phenotype was associated with decreased adhesion to HA and reduced expression of the tumor suppressor merlin in vivo. In conclusion, our study identified CD44 as a metastasis suppressor in this particular experimental OS model.

Silencing of CD44 gene expression in human 143-B osteosarcoma cells promotes metastasis of intratibial tumors in SCID mice.

The aim of this study was to characterize the different phenotypes of osteosarcoma by PET, comparing the uptake of 3 tracers ((18)F-FDG, (18)F-fluoromisonidazole [(18)F-FMISO], and (18)F-fluoride) in preclinical mouse models that reflect the heterogeneity of the human disease.

Characterization of different osteosarcoma phenotypes by PET imaging in preclinical animal models.

More effective treatment of metastasizing osteosarcoma with a current mean 5-year survival rate of less than 20% requires more detailed knowledge on mechanisms and key regulatory molecules of the complex metastatic process. CXCR4, the receptor of the chemokine CXCL12, has been reported to promote tumor progression and metastasis in osteosarcoma. CXCR7 is a recently deorphanized CXCL12-scavenging receptor with so far not well-defined functions in tumor biology. The present study focused on a potential malignancy enhancing function of CXCR7 in interaction with CXCR4 in osteosarcoma, which was investigated in an intratibial osteosarcoma model in SCID mice, making use of the human 143B osteosarcoma cell line that spontaneously metastasizes to the lung and expresses endogenous CXCR4. 143B osteosarcoma cells stably expressing LacZ (143B-LacZ cells) were retrovirally transduced with a gene encoding HA-tagged CXCR7 (143B-LacZ-X7-HA cells). 143B-LacZ-X7-HA cells co-expressing CXCR7 and CXCR4 exhibited CXCL12 scavenging and enhanced adhesion to IL-1β-activated HUVEC cells compared to 143B-LacZ cells expressing CXCR4 alone. SCID mice intratibially injected with 143B-LacZ-X7-HA cells had significantly (p<0.05) smaller primary tumors, but significantly (p<0.05) higher numbers of lung metastases than mice injected with 143B-LacZ cells. Unexpectedly, 143B-LacZ-X7-HA cells, unlike 143B-LacZ cells, also metastasized with high incidence to the auriculum cordis. In conclusion, expression of the CXCL12 scavenging receptor CXCR7 in the CXCR4-expressing human 143B osteosarcoma cell line enhances its metastatic activity in intratibial primary tumors in SCID mice that predominantly metastasize to the lung and thereby closely mimic the human disease. These findings point to CXCR7 as a target, complementary to previously proposed CXCR4, for more effective metastasis-suppressive treatment in osteosarcoma.

Expression of the chemokine receptor CXCR7 in CXCR4-expressing human 143B osteosarcoma cells enhances lung metastasis of intratibial xenografts in SCID mice.

Current combined surgical and neo-adjuvant chemotherapy of primary metastatic osteosarcoma (OS) is ineffective, reflected by a 5-year survival rate of affected patients of less than 20 %. Studies in experimental OS metastasis models pointed to the CXCR4/CXCL12 homing axis as a novel target for OS metastasis-suppressive treatment. The present study investigated for the first time the CXCR4-blocking principle in a spontaneously metastasizing human 143B OS cell line-derived orthotopic xenograft mouse model. The highly metastatic 143B cells, unlike the parental non-metastatic HOS cells, express functional CXCR4 receptors at the cell surface, as revealed in this study by RT/PCR of gene transcripts, by FACS analysis with the monoclonal anti CXCR4 antibody 12G5 (mAb 12G5) and by CXCL12 time- and dose-dependent stimulation of AKT and ERK phosphorylation. A significantly (p < 0.05) higher CXCL12 dose-dependent chemotactic response of 143B compared to HOS cells in a Boyden chamber trans-well migration assay suggested a crucial role of the CXCL12/CXCR4 homing axis in 143B cell lung metastasis. Repetitive treatment of mice with 143B cell-derived intratibial tumors given intravenous bolus injections of mAb12G5 indeed inhibited significantly (p < 0.01) the number of X-gal-stainable lung micrometastases of lacZ-transduced 143B cells. Antibody treatment had also a mild inhibitory effect on primary tumor growth associated with remarkably less osteolysis, but it did not affect the number of developing lung macrometastases. In conclusion, these results demonstrate considerable potential of high-affinity CXCR4-blocking agents for OS tumor cell homing suppressive treatment in metastasizing OS complementary to current (neo)-adjuvant chemotherapy.

Article	Year
mTHPC-mediated photodynamic therapy is effective in the metastatic human 143B osteosarcoma cells. Photochemistry and photobiology\| PubMed ID: 22268498	2012
CD44 enhances tumor formation and lung metastasis in experimental osteosarcoma and is an additional predictor for poor patient outcome. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research\| PubMed ID: 23169460	2013
Evaluation of quercetin as a potential drug in osteosarcoma treatment. Anticancer research\| PubMed ID: 23564766	2013
Silencing of CD44 gene expression in human 143-B osteosarcoma cells promotes metastasis of intratibial tumors in SCID mice. PloS one\| PubMed ID: 23565227	2013
Characterization of different osteosarcoma phenotypes by PET imaging in preclinical animal models. Journal of nuclear medicine : official publication, Society of Nuclear Medicine\| PubMed ID: 23801674	2013
Expression of the chemokine receptor CXCR7 in CXCR4-expressing human 143B osteosarcoma cells enhances lung metastasis of intratibial xenografts in SCID mice. PloS one\| PubMed ID: 24040160	2013
CXCR4 antibody treatment suppresses metastatic spread to the lung of intratibial human osteosarcoma xenografts in mice. Clinical & experimental metastasis\| PubMed ID: 24390633	2014

Carmen Campanile

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