胚胎干细胞上清液诱导Tca8113细胞发生上皮-间充质转化的实验研究

The supernatant of embryonic stem cell induces Tca8113 cell line undergoing epithelial-mesenchymal transition

目的:采用胚胎干细胞培养上清液诱导口腔鳞癌细胞发生上皮-间充质转化,初步探讨其意义。方法:应用129小鼠胚胎干细胞培养上清液诱导培养舌鳞癌Tca8113细胞,分别在1周和2周后,采用免疫荧光和免疫细胞化学检测诱导前、后Tca8113细胞中Oct4、角蛋白(CK)和波形丝蛋白(vimentin)的表达情况。半定量反转录PCR检测snail、slug、E-cadherin和CD44、Bmi-1、hTERT表达随诱导时间的变化情况。结果:Tca8113细胞经条件培养基诱导后,细胞形状逐渐由铺路石样变为长梭形;Oct4集中表达在细胞核中,vimentin转变为阳性表达,CK表达无明显变化;上皮-间充质转化标记snail mRNA表达上调,E-cadherin下调,slug无明显变化。同时,癌干细胞标记CD44、Bmi-1和hTERT mRNA表达上调。结论:口腔鳞癌细胞经胚胎干细胞培养上清液诱导后,可发生上皮-间充质转化,从而可能具备部分干细胞特性。

PURPOSE： To examine if Tca8113 cells can be induced by the supernatant of mouse embryonic stern cell to express a wider range of epithelial-mesenchymal transition （EMT） markers and stem cell markers. METHODS： Tca8113 cells were treated with recombinant supernatant of mouse embryonic stem cell and examined at 0, 1 and 2 weeks by immunohistoehemistry for Oct-4, keratin and vimentin as well as for changes in cell morphology. The reported markers of EMT： Snail, Slug and E-cadherin; and the reported cancer stem cell （CSC） markers 0aTERT, CD44 and Bmi-1） were also assessed by semi-qRT-PCR. RESULTS： TcaS113 cells changed from slabstone to spindle-shaped. Expression of Oct-4 was concentrated in cell nuclei, with up-regulation of snail, but down-regulation of E-eadherin mRNA, and only slight changes of slug. Interestingly, mRNA of hTERT, CD44 and Bmi-1 were all up-regulated. Reduced immunostaining for cytokeratin and increased vimentin confirmed the morphological transition of cells. CONCLUSIONS： Taken together, we propose that OSCC could undergo EMT acquiring partial CSC properties.