表没食子儿茶素没食子酸酯通过转化生长因子-β1／信号传导与转录激活因子-3信号通路抑制恶性黑素瘤上皮．间质转化  

Effect of epigallocatechin-3-gallate on epitheliai-mesenchymal transition in melanoma via the trans-forming growth factor-151/signal transducer and activators of transcription 3 pathway

作　　者：林云[1] 王翠彦[2] 孙兰[2] 朱里[2] 黄长征[2] 吴艳[2] 

机构地区：[1]武汉华中科技大学同济医学院附属协和医院麻醉科,430022 [2]武汉华中科技大学同济医学院附属协和医院皮肤科,430022

出　　处：《中华实验外科杂志》2013年第2期377-380,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30901984）

摘　　要：目的观察表没食子儿茶素没食子酸酯（EGCG）能否通过转化生长因子-B1（TGF-61）／信号传导与转录激活因子-3（STAT3）信号通路对恶性黑素瘤上皮-间质转化（EMT）产生影响。方法采用倒置相差显微镜观察不同浓度的EGCG（5、25、50mg／L）对TGF．p1（5ug/L）刺激的B16黑素瘤细胞作用24h后细胞形态、迁移能力和侵袭力变化。Westernblot法检测细胞磷酸化STAT3（p-STAT3）及E-钙黏蛋白（E—cadhefin）的表达。建立B16肿瘤动物模型，逆转录-聚合酶链反应（1it．PCR）检测各组瘤组织E-cadheinmRNA表达。结果（1）TGF—Bl（5ug／L）刺激B16细胞后细胞间连接不紧密，形态为长梭形，不同浓度EGCG（5、25、50mg／L）作用于TGF-61（5ug／L）刺激B16细胞24h后细胞连接较为紧密，且随浓度增加改变更加明显，细胞划痕间距逐渐增大，Transwell小室检测穿膜数依次为97．00±9．97、60．30±4．00、38．75±7．18，侵袭力与EGCG浓度呈负相关（r=-0．92，P〈0．05）。（2）不同浓度EGCG刺激组p-STAT3的表达逐渐降低，E—cadhefin表达逐渐升高，且呈浓度依赖性，p-STAT3与E．Cadhefin表达亦呈负相关（r=-0．805，P〈0．05）。（3）EGCG干预后肿瘤组织中E．cadheinmRNA水平明显增加，差异有统计学意义（P〈0．01）。结论EGCG具有明显抑制B16细胞EMT作用，其机制与抑制TGF-B1／STAT3信号通路相关。Objective To investigate the effects of epigallocatechin-3-gallate （EGCG） on epithe- lial-mesenchymal transition （EMT） through transforming growth factor （TGF）-l/signal transducer and activators of transcription 3 （ STAT3 ） signal pathway in malignant melanoma. Methods The cell morphol- ogy, migration and invasion ability of TGFq31-stimulated B16 cells were studied under a phase-contrast mi- croscope 24 h after co-incubation with different concentrations （5, 25, and 50 rag/L） of EGCG. The mi- gration ability of the cells was detected by Scrape-wound migration assay, and the invasion ability was stud- ied by Transwell. Western blotting was employed to study the expression of p-stat3 and E-cadherin. The animal model of B16 tumour was established, and E-cadhein mRNA expression was detected by using re- verse transcription-polymerase chain reaction （RT-PCR）. Results （ 1 ） TGF-I （ 5 pg/L）-stimulated B16 cells were long-spindle shaped with loose cell-cell connection. Twenty-four h later after co-incubation with different concentrations of EGCG, B16 cells became more connected, with the degree of change pro- portional to EGCG concentrations. There were increased distance in Scrape-wound migration assay, and Tr- answell number was 97. 00 ＋9. 97, 60. 30 ±4. 00 and 38.75 ±7. 18 respectively. The invasion ability was negatively correlated to the EGCG concentration （ r = - 0. 92, P 〈 0. 05 ） ; （ 2 ） The effects of reduced p- STAT3 expression and increased E-cadherin expression under the effects of different concentrations of EGCG were concentration-dependent. The expression levels of p-STATS and E-Cadherin were also nega- tively correlated （ r = - 0. 805, P 〈 0. 05 ） ; （ 3 ） The obviously increased level of E-cadhein mRNA inEGCG intervened tumour tissue had statistically significant difference （ P 〈 O. O1 ）. Conclusion EGCG in- hibits EMT of B16 cells clearly through the TGF-[31/STAT 3 signal pathway.

关 键 词：表没食子儿茶素-3-没食子酸 上皮一间质转化 转化生长因子 B1 恶性黑素瘤 

分 类 号：R73[医药卫生—肿瘤]