molecular-profiling-invasive-tumor-microenvironment-3-dimensional

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and <em>Ex vivo</em> Fibroblasts

Invading colorectal cancer (CRC) cells have acquired the capacity to break free from their sister cells, infiltrate the stroma, and remodel the ...

University of Southampton School of Medicine

BAG-1, an anti-apoptotic protein, was identified by its ability to bind to BCL-2, HSP70-family molecular chaperones and nuclear hormone receptor family members. Two BAG-1 isoforms, BAG-1L (50 kDa) and BAG-1S (32 kDa) were identified in mouse cells and BAG-1 expression was reported in murine growth plate and articular chondrocytes. The present study aimed to elucidate the role of BAG-1 in the regulation of molecular mechanisms governing chondrocyte differentiation and turnover during endochondral ossification. In long bones of skeletally immature mice, we observed expression of BAG-1 in the perichondrium, osteoblasts, osteocytes in the bone shaft, bone marrow, growth plate and articular chondrocytes. Monolayer cultures of murine chondrocytic ATDC5 cells, which exhibited robust expression of both BAG-1 isoforms and the Bag-1 transcript, were utilized as an in vitro model to delineate the roles of BAG-1. Overexpression of BAG-1L in ATDC5 cells resulted in downregulation of Col2a1 expression, a gene characteristically downregulated at the onset of hypertrophy, and an increase in transcription of Runx-2 and Alkaline phosphatase, genes normally expressed at the onset of chondrocyte hypertrophy and cartilage mineralization in the process of endochondral ossification. We also demonstrated the anti-apoptotic role of BAG-1 in chondrocytes as overexpression of BAG-1 protected ATDC5 cells, which were subjected to heat-shock at 48 degrees C for 30 min, against heat-shock-induced apoptosis. Overexpression of the SOX-9 protein in ATDC5 cells resulted in increased Bag-1 gene expression. To further investigate the regulation of Bag-1 gene expression by SOX-9, CHO cells were co-transfected with the human Bag-1 gene promoter-Luciferase reporter construct and the human pSox-9 expression vector. Activity of the Bag-1 promoter was significantly enhanced by the SOX-9 protein. In conclusion, a novel finding of this study is the role of BAG-1 as a transcriptional regulator of genes involved in chondrocyte hypertrophy and cartilage mineralization during the process of endochondral ossification. Additionally, we have demonstrated for the first time the regulation of Bag-1 gene expression by SOX-9 and the anti-apoptotic role of BAG-1 in chondrocytic cells. Modulation of Bag-1 expression can therefore mediate chondrocyte differentiation and turnover, and offer further insight into the molecular regulation of endochondral ossification.

Bcl-2-associated athanogene-1 (BAG-1): a transcriptional regulator mediating chondrocyte survival and differentiation during endochondral ossification.

&#x6297; CD20 &#x5355;&#x514B;&#x9686;&#x6297;&#x4F53; (&#x5355;&#x514B;&#x9686;&#x6297;&#x4F53;) &#x5229;&#x59A5;&#x6614;&#x5355;&#x6297;&#x73B0;&#x5728;&#x7ECF;&#x5E38;&#x7528;&#x4E8E;&#x6CBB;&#x7597;&#x975E;&#x970D;&#x5947;&#x91D1;&#x6DCB;&#x5DF4;&#x7624;&#x548C;&#x73B0;&#x6B63;&#x8FDB;&#x884C;&#x5E7F;&#x6CDB;&#x7684;&#x5176;&#x4ED6; B &#x7EC6;&#x80DE;&#x75BE;&#x75C5;&#xFF0C;&#x5982;&#x7C7B;&#x98CE;&#x6E7F;&#x6027;&#x5173;&#x8282;&#x708E;&#x3002;&#x6DF1;&#x5165;&#x7EC6;&#x81F4;&#x7684;&#x7814;&#x7A76;&#xFF0C;&#x5C3D;&#x7BA1;&#x4F5C;&#x7528;&#x673A;&#x7406;&#x4ECD;&#x7136;&#x662F;&#x4E0D;&#x786E;&#x5B9A;&#x3002;&#x5728;&#x5F53;&#x524D;&#x7684;&#x7814;&#x7A76;&#x4E2D;&#xFF0C;&#x8C03;&#x67E5;&#x4E86;&#x6297; CD20 &#x5355;&#x514B;&#x9686;&#x6297;&#x4F53;&#x8BF1;&#x5BFC;&#x9499;&#x4FE1;&#x53F7;&#x3002;&#x4EE5;&#x524D;&#xFF0C;&#x6211;&#x4EEC;&#x5206;&#x4E3A;&#x6297; CD20 &#x5355;&#x514B;&#x9686;&#x6297;&#x4F53;&#x7C7B;&#x578B;&#x6839;&#x636E;&#x5404;&#x79CD;&#x7279;&#x5F81;&#xFF0C;&#x5982;&#x4ED6;&#x4EEC;&#x7684;&#x80FD;&#x529B;&#xFF0C;&#x4EE5;&#x8BF1;&#x4F7F;&#x8865;&#x4F53;&#x6FC0;&#x6D3B;&#x548C;&#x91CD;&#x65B0; CD20 &#x5206;&#x53D1;&#x5230;&#x6D17;&#x6DA4;&#x5242;&#x6EB6;&#x819C;&#x57DF;&#x6211; (&#x5229;&#x59A5;&#x6614;&#x5355;&#x6297;&#x6837;) &#x548C; II &#x578B; &#xFF08;B1-&#x50CF;&#xFF09;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#x6211;&#x4EEC;&#x663E;&#x793A;&#x6211;&#x5355;&#x514B;&#x9686;&#x6297;&#x4F53;&#x80FD;&#x591F;&#x8BF1;&#x5BFC; B &#x7EC6;&#x80DE;&#x7684;&#x9499;&#x901A;&#x548C;&#xFF0C;&#x8FD9;&#x4E0E;&#x8868;&#x8FBE;&#x7684; B &#x7EC6;&#x80DE;&#x6297;&#x539F;&#x53D7;&#x4F53; (BCR) &#x5BC6;&#x5207;&#x76F8;&#x5173;&#x7684;&#x552F;&#x4E00;&#x7C7B;&#x578B;&#x3002;&#x6291;&#x5236;&#x5242;&#x5206;&#x6790;&#x663E;&#x793A;&#x96C7;&#x7528;&#x7684; CD20 &#x7684;&#x4FE1;&#x53F7;&#x7EA7;&#x8054;&#x4E0E;&#x4F7F;&#x7528;&#x7684; BCR Syk&#x3001; Src&#xFF0C;&#x548C;&#x808C;&#x9187;&#xFF0C;&#x4F46;&#x4E0D;&#x662F; EGTA&#xFF0C;p38 &#x7684;&#x6291;&#x5236;&#x5242;&#x6216; ERK1/2&#xFF0C;&#x5B8C;&#x5168;&#x70E7;&#x8680;&#x9499;&#x901A;&#x91CF;&#x60CA;&#x4EBA;&#x5730;&#x76F8;&#x4F3C;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x7ED1;&#x5B9A;&#x7C7B;&#x578B;&#x7684;&#x6211;&#xFF0C;&#x4F46;&#x4E0D;&#x662F;&#x4E0E;&#x4F5C;&#x4E3A; BCR &#x9020;&#x6210;&#x76F4;&#x63A5;&#x5173;&#x8054;&#x7684; CD20 II &#x578B;&#x5355;&#x514B;&#x9686;&#x6297;&#x4F53;&#x6D4B;&#x5B9A;&#x97F3;&#x54C1;&#xFF0C;&#x5BFC;&#x81F4; BLNK BCR &#x7279;&#x5B9A;&#x9002;&#x914D;&#x5668;&#x86CB;&#x767D;&#x8D28;&#x7684;&#x78F7;&#x9178;&#x5316;&#x548C; SLP-76&#x3002;&#x81F3;&#x5173;&#x91CD;&#x8981;&#x7684;&#x662F;&#xFF0C;&#x62C9;&#x83AB;&#x65AF;&#x53D8;&#x5F02;&#x7EC6;&#x80DE;&#x7F3A;&#x4E4F; BCR &#x8868;&#x8FBE;&#x4F46;&#x4E0D;&#x53D8;&#x7684; CD20 &#x4E0E;&#x8868;&#x8FBE;&#x4E86;&#x5B8C;&#x5168;&#x65E0;&#x6CD5;&#x4FC3;&#x4F7F;&#x9499;&#x52A9;&#x710A;&#x5242; CD20 &#x7ED3;&#x624E;&#x540E;&#x3002;&#x96C6;&#x4F53;&#xFF0C;&#x8FD9;&#x4E9B;&#x6570;&#x636E;&#x8868;&#x660E; CD20 &#x8BF1;&#x5BFC;&#x9499;&#x901A;&#x901A;&#x8FC7;&#x5173;&#x8054;&#x548C;&quot;&#x52AB;&#x6301;&quot;&#x4FE1;&#x53F7;&#x7684; BCR &#x6F5C;&#x529B;&#x7684;&#x80FD;&#x529B;&#x3002;

&#x611F;&#x5E94;&#x7684;&#x6E38;&#x79BB;&#x9499;&#x79BB;&#x5B50;&#x901A;&#x91CF;&#x7684; CD20 &#x662F;&#x53D6;&#x51B3;&#x4E8E; B &#x7EC6;&#x80DE;&#x6297;&#x539F;&#x53D7;&#x4F53;&#x4FE1;&#x53F7;&#x3002;

The anti-CD20 monoclonal antibody (mAb) rituximab is now routinely used for the treatment of non-Hodgkins lymphoma and is being examined in a wide range of other B-cell disorders, such as rheumatoid arthritis. Despite intensive study, the mechanism of action still remains uncertain. In the current study, anti-CD20 mAb-induced calcium signaling was investigated. Previously, we grouped anti-CD20 mAbs into Type I (rituximab-like) and Type II (B1-like) based upon various characteristics such as their ability to induce complement activation and redistribute CD20 into detergent-insoluble membrane domains. Here we show that only Type I mAbs are capable of inducing a calcium flux in B cells and that this is tightly correlated with the expression of the B-cell antigen receptor (BCR). Inhibitor analysis revealed that the signaling cascade employed by CD20 was strikingly similar to that utilized by the BCR, with inhibitors of Syk, Src, and PI3K, but not EGTA, p38, or ERK1/2, completely ablating calcium flux. Furthermore, binding of Type I but not Type II mAbs caused direct association of CD20 with the BCR as measured by FRET and resulted in the phosphorylation of BCR-specific adaptor proteins BLNK and SLP-76. Crucially, variant Ramos cells lacking BCR expression but with unchanged CD20 expression were completely unable to induce calcium flux following ligation of CD20. Collectively, these data indicate that CD20 induces cytosolic calcium flux through its ability to associate with and "hijack" the signaling potential of the BCR.

Induction of cytosolic calcium flux by CD20 is dependent upon B Cell antigen receptor signaling.

Le rituximab d&#39;anticorps monoclonal (ACM) anti-CD20 est maintenant couramment utilis&eacute; pour le traitement du lymphome non hodgkinien et est examin&eacute;e dans un large &eacute;ventail d&#39;autres troubles de B-cellule, comme la polyarthrite rhumato&iuml;de. Malgr&eacute; des &eacute;tudes intensives, le m&eacute;canisme d&#39;action reste encore incertain. Dans la pr&eacute;sente &eacute;tude, anti-CD20 de signalisation calcique induite par le mAb a &eacute;t&eacute; &eacute;tudi&eacute;e. Auparavant, nous avons regroup&eacute; les ACM anti-CD20 en Type I (comme le rituximab) et Type II (type B1) selon diverses caract&eacute;ristiques telles que leur capacit&eacute; &agrave; induire l&#39;activation du compl&eacute;ment et redistribuer CD20 dans domaines membranaires insolubles dans un d&eacute;tergent. Ici, nous montrons que seul Type j&#39;ai mAbs sont capables d&#39;induire un flux de calcium dans les cellules de B, et que cela est &eacute;troitement corr&eacute;l&eacute;e avec l&#39;expression du r&eacute;cepteur de l&#39;antig&egrave;ne des cellules B (BCR). Inhibiteur analyse a r&eacute;v&eacute;l&eacute; que la cascade de signalisation employ&eacute;e par CD20 remarquablement semblable &agrave; celle utilis&eacute;e par la BCR, avec des inhibiteurs de Syk, Src et PI3K, mais pas l&#39;EGTA, p38, ou ERK1/2, ablation compl&egrave;tement le flux de calcium. En outre, j&#39;ai, mais pas de Type II mAbs caus&eacute; une association directe du CD20 avec la RCB comme la liaison de Type mesur&eacute;e par frette et entra&icirc;n&eacute; la phosphorylation des prot&eacute;ines d&#39;adaptateur sp&eacute;cifique BCR BLNK et SLP-76. C&#39;est essentiel, variante Ramos cellules manque expression BCR, mais le CD20 inchang&eacute;e expression furent compl&egrave;tement incapables d&#39;induire le flux de calcium apr&egrave;s ligature de CD20. Ensemble, ces donn&eacute;es indiquent que le CD20 induit le flux de calcium cytosolique gr&acirc;ce &agrave; sa capacit&eacute; d&#39;associer &agrave; &laquo; d&eacute;tourner &raquo; le potentiel de signalisation de la BCR.

Induction du flux de calcium cytosolique de CD20 est tributaire de la signalisation de r&eacute;cepteur de l&#39;antig&egrave;ne B Cell.

El rituximab del anticuerpo monoclonal (mAb) anti-CD20 ahora se utiliza habitualmente para el tratamiento del linfoma no-Hodgkin y se examina en una amplia gama de otros trastornos de c&eacute;lulas B, como la artritis reumatoide. A pesar de estudio intensivo, el mecanismo de acci&oacute;n sigue siendo incierto. En el presente estudio, se investig&oacute; se&ntilde;alizaci&oacute;n de calcio inducida por mAb anti-CD20. Previamente, se agruparon los mAbs anti-CD20 en tipo I (rituximab-como) y tipo II (B1-como) basado en diversas caracter&iacute;sticas tales como su capacidad para inducir la activaci&oacute;n del complemento y redistribuir CD20 en dominios de membrana insoluble en detergente. Aqu&iacute; demostramos que &uacute;nico tipo me mAbs son capaces de inducir un flujo de calcio en las c&eacute;lulas de B y que esto se correlaciona estrechamente con la expresi&oacute;n del receptor del ant&iacute;geno de c&eacute;lulas B (BCR). Inhibidor an&aacute;lisis revel&oacute; que la cascada de se&ntilde;alizaci&oacute;n empleada por CD20 era sorprendentemente similar a la utilizada por el BCR, con inhibidores de Syk, Src y PI3K, pero no EGTA, p38, o ERK1/2, ablaci&oacute;n completamente el flujo de calcio. Adem&aacute;s, atascamiento del tipo I pero no del tipo II mAbs causada directa Asociaci&oacute;n de CD20 con el BCR como medido por traste y dio lugar a la fosforilaci&oacute;n de prote&iacute;nas del adaptador del BCR espec&iacute;ficas BLNK y SLP-76. Crucial, variante Ramos las c&eacute;lulas expresi&oacute;n de BCR pero con CD20 sin cambiar la expresi&oacute;n era totalmente incapaz de inducir el flujo de calcio despu&eacute;s de la ligadura de CD20. Colectivamente, estos datos indican que CD20 induce el flujo de calcio citos&oacute;lico a trav&eacute;s de su capacidad de asociar y &quot;secuestrar&quot; el potencial de la se&ntilde;alizaci&oacute;n de la BCR.

Inducci&oacute;n del flujo de calcio citos&oacute;lico por CD20 depende de c&eacute;lulas B ant&iacute;geno receptor se&ntilde;alizaci&oacute;n.

Die Anti-CD20 monoklonalen Antik&ouml;rper (mAb) Rituximab ist jetzt f&uuml;r die Behandlung des non-Hodgkins Lymphoms routinem&auml;&szlig;ig verwendet und wird in einer Vielzahl von anderen B-Zell-Erkrankungen wie rheumatoide Arthritis gepr&uuml;ft. Trotz intensiver Studien bleibt der Wirkmechanismus noch ungewiss. In der aktuellen Studie wurde die Anti-CD20 mAb-induzierte Kalzium Signalisierung untersucht. Zuvor, gruppiert wir Anti-CD20-mAbs in Typ I (Rituximab-&auml;hnlich) und Typ II (B1-&auml;hnlich) basierend auf verschiedene Merkmale wie ihre F&auml;higkeit, induzieren Komplement-Aktivierung und CD20 in Waschmittel-unl&ouml;slich Membran-Dom&auml;nen zu verteilen. Hier zeigen wir, dass nur Typ I mAbs induzieren eine Kalziumflu&szlig; in B-Zellen f&auml;hig sind und dies ist eng korreliert mit dem Ausdruck von der B-Zell-Antigen-Rezeptor (BCR). Inhibitor-Analyse ergab, dass die signalisierende Kaskade von CD20 besch&auml;ftigt &Auml;hnlichkeit mit der von der BCR mit Inhibitoren der Syk, Src, und PI3K, aber nicht EGTA, p38, oder ERK1/2, komplett Abtragen Kalziumflu&szlig; eingesetzt. Dar&uuml;ber hinaus verbindliche Art, ich aber nicht Typ II mAbs verursachte direkte Assoziation von CD20 mit BCR als Bund gemessen und resultierte in der Phosphorylierung von BCR-spezifische Adapter Proteine BLNK und SLP-76. Entscheidend ist, Variante Ramos Zellen fehlt BCR Ausdruck mit unver&auml;ndert CD20 Ausdruck konnten aber komplett nach Verbindung der CD20 Kalziumflu&szlig; veranlassen. Diese Daten zufolge gemeinsam CD20 induziert Zytosolische Kalziumflu&szlig; durch seine F&auml;higkeit, zuordnen und die &quot;Entf&uuml;hrung&quot; das Signalling Potenzial des BCR.

Induktion von Zytosolische Kalziumflu&szlig; von CD20 ist abh&auml;ngig von B-Zell-Antigen-Rezeptor-Signalisierung.

Il rituximab anticorpo monoclonale (mAb) anti-CD20 &egrave; utilizzati di routine per il trattamento del linfoma non-Hodgkins e viene esaminato in una vasta gamma di altri disturbi della B-cellula, come l&#39;artrite reumatoide. Nonostante lo studio intenso, il meccanismo d&#39;azione rimane ancora incerto. Nello studio corrente, anti-CD20 mAb-indotta calcio segnalazione &egrave; stato indagato. In precedenza, abbiamo raggruppati mAbs anti-CD20 in tipo I (rituximab-like) e tipo II (B1-like) basato su varie caratteristiche come la loro capacit&agrave; di indurre l&#39;attivazione del complemento e ridistribuire CD20 in domini di membrana detergente insolubile. Qui ci mostrano che solo tipo I mAbs sono in grado di indurre un flusso di calcio nelle cellule B e che ci&ograve; &egrave; strettamente correlato con l&#39;espressione del recettore dell&#39;antigene delle cellule di B (BCR). Inibitore analisi hanno rivelato che la cascata di segnalazione impiegata da CD20 era sorprendentemente simile a quello utilizzato da BCR, con inibitori di Syk, Src e PI3K, ma non EGTA, p38, o ERK1/2, ablazione completamente il flusso di calcio. Inoltre, binding di tipo io, ma non il tipo II mAbs causato associazione diretta di CD20 con BCR come misurato da FRET e ha portato la fosforilazione delle proteine di adattatore specifico BCR BLNK e SLP-76. Fondamentalmente, variante Ramos cellule carente espressione di BCR ma con immutata CD20 espressione erano completamente in grado di indurre il flusso di calcio dopo legatura di CD20. Collettivamente, questi dati indicano che CD20 induce il flusso di calcio citosolico attraverso la sua capacit&agrave; di associare e di &quot;dirottare&quot; il potenziale segnalazione del BCR.

Induzione del flusso di calcio citosolico di CD20 &egrave; dipendente delle cellule di B antigene recettore segnalazione.

&#x6297; CD20 &#x30E2;&#x30CE;&#x30AF;&#x30ED;&#x30FC;&#x30CA;&#x30EB;&#x6297;&#x4F53; (mAb) &#x30EA;&#x30C4;&#x30AD;&#x30B7;&#x30DE;&#x30D6;&#x306F;&#x4ECA;&#x5B9A;&#x671F;&#x7684;&#x306B;&#x975E;&#x30DB;&#x30B8;&#x30AD;&#x30F3; &#x30EA;&#x30F3;&#x30D1;&#x816B;&#x306E;&#x6CBB;&#x7642;&#x306B;&#x5229;&#x7528;&#x3055;&#x308C;&#x307E;&#x3059;&#x3001;&#x30EA;&#x30A6;&#x30DE;&#x30C1;&#x6027;&#x95A2;&#x7BC0;&#x708E;&#x306A;&#x3069;&#x306E;&#x4ED6;&#x306E; B &#x7D30;&#x80DE;&#x969C;&#x5BB3;&#x306E;&#x5E83;&#x3044;&#x7BC4;&#x56F2;&#x3067;&#x691C;&#x8A0E;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x96C6;&#x4E2D;&#x7684;&#x306A;&#x7814;&#x7A76;&#x306B;&#x3082;&#x304B;&#x304B;&#x308F;&#x3089;&#x305A;&#x3001;&#x4F5C;&#x7528;&#x306E;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x306F;&#x307E;&#x3060;&#x4E0D;&#x900F;&#x660E;&#x3067;&#x3059;&#x3002;&#x73FE;&#x5728;&#x306E;&#x7814;&#x7A76;&#x3067;&#x306F;, &#x6297; CD20 &#x6297;&#x4F53;&#x306B;&#x3088;&#x308B;&#x30AB;&#x30EB;&#x30B7;&#x30A6;&#x30E0; &#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x3092;&#x8ABF;&#x3079;&#x305F;&#x3002;&#x4EE5;&#x524D;&#x306F;&#x3001;&#x6211;&#x3005; &#x6297; CD20 &#x30E2;&#x30CE;&#x30AF;&#x30ED;&#x30FC;&#x30CA;&#x30EB;&#x6297;&#x4F53;&#x306E;&#x7A2E;&#x985E;&#x306B;&#x30B0;&#x30EB;&#x30FC;&#x30D7;&#x5316; I (&#x30EA;&#x30C4;&#x30AD;&#x30B7;&#x30DE;&#x30D6;&#x306E;&#x3088;&#x3046;&#x306A;&#xFF09; &#x3068;&#x30BF;&#x30A4;&#x30D7; II &#xFF08;B1 &#x306E;&#x3088;&#x3046;&#x306A;) &#x88DC;&#x4F53;&#x6D3B;&#x6027;&#x5316;&#x3092;&#x8A98;&#x5C0E;&#x3057;&#x3001;CD20 &#x6D17;&#x5264;&#x4E0D;&#x6EB6;&#x6027;&#x819C;&#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x306B;&#x518D;&#x914D;&#x5E03;&#x3059;&#x308B;&#x80FD;&#x529B;&#x306A;&#x3069;&#x306E;&#x3055;&#x307E;&#x3056;&#x307E;&#x306A;&#x7279;&#x6027;&#x306B;&#x57FA;&#x3065;&#x3044;&#x3066;&#x3002;&#x3053;&#x3053;&#x3067;&#x79C1;&#x9054;&#x306F; Mab &#x306F; B &#x7D30;&#x80DE;&#x306E;&#x30AB;&#x30EB;&#x30B7;&#x30A6;&#x30E0;&#x306E;&#x30D5;&#x30E9;&#x30C3;&#x30AF;&#x30B9;&#x3092;&#x8A98;&#x5C0E;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x308B;&#x3001;&#x3053;&#x306E;&#x3057;&#x3063;&#x304B;&#x308A;&#x3068; B &#x7D30;&#x80DE;&#x6297;&#x539F;&#x53D7;&#x5BB9;&#x4F53; &#xFF08;BCR&#xFF09; &#x306E;&#x767A;&#x73FE;&#x3068;&#x76F8;&#x95A2;&#x3057;&#x3066;&#x3044;&#x308B;&#x552F;&#x4E00;&#x306E;&#x30BF;&#x30A4;&#x30D7;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x963B;&#x5BB3;&#x5264;&#x89E3;&#x6790;&#x306F;&#x3001;CD20 &#x3067;&#x63A1;&#x7528;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x30AB;&#x30B9;&#x30B1;&#x30FC;&#x30C9;&#x306F;&#x9A5A;&#x304F;&#x307B;&#x3069;&#x4F3C;&#x3066;&#x3044;&#x306A;&#x3044; egta &#x51E6;&#x7406;&#x306B;&#x3088;&#x308A;&#x8457;&#x3057;&#x304F;&#x3001;PI3K&#x3001;Src&#x3001;Syk p38 &#x306E;&#x963B;&#x5BB3;&#x5264;&#x3068;&#x306E;&#x7403;&#x6D77;&#x7DBF;&#x4F53;&#x307E;&#x305F;&#x306F; ERK1/2 &#x5B8C;&#x5168;&#x306B;&#x30AB;&#x30EB;&#x30B7;&#x30A6;&#x30E0;&#x306E;&#x30D5;&#x30E9;&#x30C3;&#x30AF;&#x30B9;&#x3092;&#x30A2;&#x30D6;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306B;&#x3088;&#x308B;&#x5229;&#x7528;&#x306B;&#x3042;&#x3063;&#x305F;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x30D0;&#x30A4;&#x30F3;&#x30C9;&#x306E;&#x7A2E;&#x985E;&#x79C1;&#x304C;&#x306A;&#x3044;&#x30BF;&#x30A4;&#x30D7; II Mab &#x306E;&#x76F4;&#x63A5;&#x5354;&#x4F1A; CD20 &#x7403;&#x6D77;&#x7DBF;&#x4F53;&#x3068;&#x3057;&#x3066;&#x767A;&#x751F;&#x30D5;&#x30EC;&#x30C3;&#x30C8;&#x3067;&#x6E2C;&#x5B9A;&#x3057;&#x3001;BCR &#x7279;&#x5B9A;&#x30A2;&#x30C0;&#x30D7;&#x30BF;&#x30FC;&#x86CB;&#x767D;&#x8CEA; BLNK &#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x3068; SLP 76 &#x306E;&#x7D50;&#x679C;&#x3057;&#x307E;&#x3059;&#x3002;&#x6C7A;&#x5B9A;&#x7684;&#x306B;&#x3001;&#x30D0;&#x30EA;&#x30A2;&#x30F3;&#x30C8; &#x30E9;&#x30E2;&#x30B9;&#x6B20;&#x3051;&#x3066;&#x3044;&#x308B; BCR &#x5F0F;&#x7D30;&#x80DE;&#x304C;&#x3001;&#x5909;&#x66F4;&#x306E; CD20 &#x3068;&#x5F0F;&#x306F;&#x5B8C;&#x5168;&#x306B;&#x6B21;&#x306E;&#x7D50;&#x7D2E; CD20 &#x306E;&#x30AB;&#x30EB;&#x30B7;&#x30A6;&#x30E0;&#x5909;&#x5316;&#x3092;&#x8A98;&#x767A;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x305B;&#x3093;&#x3067;&#x3057;&#x305F;&#x3002;&#x7DCF;&#x79F0;&#x3057;&#x3066;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x30C7;&#x30FC;&#x30BF;&#x306F;&#x3001;cd20 &#x967D;&#x6027;&#x7D30;&#x80DE;&#x8CEA;&#x30AB;&#x30EB;&#x30B7;&#x30A6;&#x30E0;&#x306E;&#x30D5;&#x30E9;&#x30C3;&#x30AF;&#x30B9;&#x3092;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x308B;&#x3057;&#x3001;&#x300C;BCR &#x306E;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x306E;&#x53EF;&#x80FD;&#x6027;&#x3092;&#x30CF;&#x30A4;&#x30B8;&#x30E3;&#x30C3;&#x30AF;&#x300D;&#x3059;&#x308B;&#x80FD;&#x529B;&#x3092;&#x8A98;&#x5C0E;&#x3059;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;

Cd20 &#x967D;&#x6027;&#x7D30;&#x80DE;&#x8CEA;&#x30AB;&#x30EB;&#x30B7;&#x30A6;&#x30E0; &#x30D5;&#x30E9;&#x30C3;&#x30AF;&#x30B9;&#x306E;&#x8A98;&#x5C0E;&#x306F; B &#x7D30;&#x80DE;&#x6297;&#x539F;&#x53D7;&#x5BB9;&#x4F53;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x306B;&#x4F9D;&#x5B58;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;

&#xC548;&#xD2F0;-CD20 &#xB2E8;&#xC77C; &#xD074;&#xB860; &#xD56D; &#xCCB4; (mAb) rituximab &#xC9C0;&#xAE08; &#xC77C;&#xC0C1;&#xC801;&#xC73C;&#xB85C; &#xBE44; Hodgkins &#xB9BC;&#xD504; &#xC885; &#xCE58;&#xB8CC;&#xC5D0; &#xC0AC;&#xC6A9; &#xB418;&#xB294; &#xBC0F; &#xB2E4;&#xB978; B &#xC138;&#xD3EC; &#xC7A5;&#xC560;, &#xB958; &#xB9C8;&#xD2F0;&#xC2A4; &#xAD00;&#xC808;&#xC5FC; &#xB4F1;&#xC758; &#xB113;&#xC740; &#xBC94;&#xC704;&#xC5D0;&#xC11C; &#xAC80;&#xC0AC;&#xB97C; &#xBC1B;&#xACE0;. &#xC9D1;&#xC911;&#xC801;&#xC778; &#xC5F0;&#xAD6C;&#xC5D0;&#xB3C4; &#xBD88;&#xAD6C; &#xD558; &#xACE0; &#xB3D9;&#xC791;&#xC758; &#xBA54;&#xCEE4;&#xB2C8;&#xC998;&#xC774; &#xC5EC;&#xC804;&#xD788; &#xBD88;&#xD655;&#xC2E4; &#xD558;&#xB2E4;. &#xD604;&#xC7AC; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; &#xC548;&#xD2F0;-CD20 mAb &#xC720;&#xB3C4; &#xCE7C;&#xC298; &#xC2E0;&#xD638; &#xC870;&#xC0AC; &#xD588;&#xB2E4;. &#xC774;&#xC804;&#xC5D0;, &#xC6B0;&#xB9AC;&#xB294; &#xD615;&#xC2DD;&#xC73C;&#xB85C; &#xC548;&#xD2F0;-CD20 mAbs &#xADF8;&#xB8F9;&#xD654; (rituximab &#xAC19;&#xC740;) &#xBC0F; &#xC720;&#xD615; II (B1 &#xAC19;&#xC740;) &#xADF8;&#xB4E4;&#xC758; &#xB2A5;&#xB825;&#xC744; &#xBCF4;&#xC644; &#xD65C;&#xC131;&#xD654;&#xB97C; &#xC720;&#xB3C4; &#xD558; &#xACE0; &#xC138;&#xC81C; &#xBD88;&#xC6A9; &#xC131; &#xB9C9; &#xB3C4;&#xBA54;&#xC778;&#xC73C;&#xB85C; CD20 &#xC7AC;&#xBC30;&#xD3EC; &#xB4F1;&#xC758; &#xB2E4;&#xC591; &#xD55C; &#xD2B9;&#xC131;&#xC5D0; &#xB530;&#xB77C;. &#xC5EC;&#xAE30; &#xC6B0;&#xB9AC;&#xAC00; &#xD558;&#xB294; &#xC720;&#xC77C;&#xD55C; &#xC720;&#xD615; &#xB0B4;&#xAC00; mAbs B &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xCE7C;&#xC298; &#xD50C;&#xB7ED;&#xC2A4;&#xB97C; &#xC720;&#xB3C4; &#xD560; &#xC218; &#xC788;&#xB2E4; &#xADF8;&#xB9AC;&#xACE0; &#xC774;&#xAC83;&#xC740; &#xBC00;&#xC811; &#xD558; &#xAC8C; B &#xC138;&#xD3EC; &#xD56D; &#xC6D0; &#xC218;&#xC6A9; &#xCCB4; (BCR) &#xC2DD;&#xC640; &#xC0C1;&#xAD00;&#xC744; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. &#xC5B5;&#xC81C;&#xC81C; &#xBD84;&#xC11D; &#xACF5;&#xAC1C; CD20 &#xACE0;&#xC6A9; &#xC2E0;&#xD638; &#xCE90;&#xC2A4;&#xCF00;&#xC774;&#xB4DC; Syk, Src, &#xBC0F; PI3K, &#xD558;&#xC9C0;&#xB9CC; &#xD558;&#xC9C0; EGTA p38 &#xC5B5;&#xC81C;&#xC81C;&#xC640; BCR &#xB610;&#xB294; ERK1/2, &#xC644;&#xC804; &#xD558; &#xAC8C; ablating &#xCE7C;&#xC298; &#xD50C;&#xB7ED;&#xC2A4; &#xB110;&#xB9AC; &#xC0AC;&#xC6A9; &#xAE30; &#xB9C9;&#xD788;&#xAC8C; &#xBE44;&#xC2B7; &#xD588;&#xB2E4;. &#xB610;&#xD55C;, &#xB09C; &#xD558;&#xC9C0;&#xB9CC; CD20 &#xC9C1;&#xC811; &#xD611;&#xD68C;&#xB85C; BCR &#xBC1C;&#xC0DD; &#xD558;&#xC9C0; &#xC720;&#xD615; II mAbs &#xD615;&#xC2DD;&#xC758; &#xBC14;&#xC778;&#xB529; &#xBB34;&#xC11C; &#xC6CC; &#xD558; &#xC5EC; &#xCE21;&#xC815; &#xD558; &#xACE0; &#xADC0;&#xCC29;&#xB418; &#xC5C8; &#xB2E4; BCR &#xD2B9;&#xC815; BLNK &#xC5B4;&#xB311;&#xD130; &#xB2E8;&#xBC31;&#xC9C8;&#xC758; &#xC778; &#xC0B0;&#xD654;&#xC640; SLP-76. &#xACB0;&#xC815;&#xC801;&#xC778;, &#xBCC0;&#xC885; &#xB77C;&#xBAA8;&#xC2A4; &#xC138;&#xD3EC; &#xBD80;&#xC871; BCR &#xC2DD; &#xD558;&#xC9C0;&#xB9CC; &#xADF8;&#xB300;&#xB85C; cd20&#xC640; &#xC2DD; &#xC644;&#xC804;&#xD788; &#xC218; &#xCE7C;&#xC298; &#xD50C;&#xB7ED;&#xC2A4; cd20&#xC758; &#xACB0; &#xCC30;&#xC5D0; &#xB530;&#xB77C; &#xC720;&#xB3C4; &#xB418;&#xC9C0; &#xC54A;&#xC558;&#xB2E4;. &#xC0E8; &#xB2E4;, &#xC774;&#xB7EC;&#xD55C; &#xB370;&#xC774;&#xD130; CD20 &#xC720;&#xB3C4; cytosolic &#xCE7C;&#xC298; &#xC720;&#xB3D9;&#xC801;&#xC73C;&#xB85C; &#xC5F0;&#xACB0; &#xD558; &#xACE0; &quot;&#xACF5;&#xC911; &#xB0A9;&#xCE58;&quot;&#xB294; BCR&#xC758; &#xC2E0;&#xD638; &#xC7A0;&#xC7AC;&#xB825;&#xC744; &#xD1B5;&#xD574; &#xB098;&#xD0C0;&#xB0C5;&#xB2C8;&#xB2E4;.

CD20 &#xC5EC; cytosolic &#xCE7C;&#xC298; &#xD50C;&#xB7ED;&#xC2A4;&#xC758; &#xC720;&#xB3C4; B &#xC138;&#xD3EC; &#xD56D; &#xC6D0; &#xC218;&#xC6A9; &#xCCB4; &#xC2E0;&#xD638;&#xC5D0; &#xB530;&#xB77C; &#xB2E4;&#xB985;&#xB2C8;&#xB2E4;.

MicroRNAs (miRNAs) are a class of small highly conserved RNAs that provide widespread expressional control through the translational repression of mRNA. MiRNAs have fundamental roles in the regulation of intracellular processes, and their importance during malignant transformation and metastasis is becoming increasingly well recognized. An important event in the metastatic cascade is epithelial to mesenchymal transition (EMT), a reversible phenotypic switch over, which endows malignant epithelial cells with the capacity to break free from one another and invade the surrounding stroma. Our understanding of EMT has been significantly improved by the characterization of miRNAs that influence the signalling pathways and downstream events that define EMT on a molecular level. Here, we detail the role of miRNAs in EMT, and in doing so demonstrate their importance in the early stages of the metastatic cascade; we discuss a significant body of data that suggest new opportunities for drug development, and we highlight critical knowledge gaps that remain to be addressed.

MicroRNAs: critical regulators of epithelial to mesenchymal (EMT) and mesenchymal to epithelial transition (MET) in cancer progression.

Hypoxia inducible factor-1 (HIF-1) is a heterodimeric transcription factor that acts as the master regulator of cellular response to reduced oxygen levels, thus playing a key role in the adaptation, survival, and progression of tumors. Here we report cyclo-CLLFVY, identified from a library of 3.2 million cyclic hexapeptides using a genetically encoded high-throughput screening platform, as an inhibitor of the HIF-1α/HIF-1β protein-protein interaction in vitro and in cells. The identified compound inhibits HIF-1 dimerization and transcription activity by binding to the PAS-B domain of HIF-1α, reducing HIF-1-mediated hypoxia response signaling in a variety of cell lines, without affecting the function of the closely related HIF-2 isoform. The reported cyclic peptide demonstrates the utility of our high-throughput screening platform for the identification of protein-protein interaction inhibitors, and forms the starting point for the development of HIF-1 targeted cancer therapeutics.

Graham K. Packham

Cancer Sciences Unit

University of Southampton School of Medicine

Molecular Profiling of the Invasive Tumor Microenvironment in a 3-Dimensional Model of Colorectal Cancer Cells and Ex vivo Fibroblasts