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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。
OBJECTIVE: To induce epithelial-mesenchymal transition in squamous cell carcinoma of the oral squamous cell carcinoma using embryonic stem cell culture supernatant to explore its significance. Methods: Cultured human tongue squamous cell carcinoma Tca8113 cells using culture supernatant of 129 mouse embryonic stem cells. Immunofluorescence and immunocytochemistry were used to detect the expression of Oct4 and keratin (CK) in Tca8113 cells after 1 week and 2 weeks respectively ) And the expression of vimentin. Semi-quantitative reverse transcription PCR was used to detect the changes of snail, slug, E-cadherin and CD44, Bmi-1 and hTERT with the induction time. Results: Tca8113 cells were transformed into long fusiform shape by paving-stone-like cells induced by conditioned medium. The expression of Oct4 in the nucleus was positive, and the expression of vimentin was positive but the expression of CK was unchanged. The expression of epithelial-mesenchymal transition Snail mRNA expression was up-regulated, E-cadherin down-regulated, slug no significant change. Meanwhile, the expression of cancer stem cell markers CD44, Bmi-1 and hTERT mRNA were up-regulated. CONCLUSIONS: Oral squamous cell carcinoma cells may undergo epithelial-mesenchymal transition induced by ES cell culture supernatant, which may have some stem cell characteristics.