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目的:探讨Klf4表达水平对Hela细胞上皮间质转化的影响。方法:设计合成针对Klf4的siRNA和阴性对照siRNA,并转染至Hela细胞。用含10ng/ml TGF-β1和10%胎牛血清的DMEM培养液诱导Hela细胞发生上皮间质转化,对照组使用不含TGF-β1的培养液。倒置显微镜下观察记录细胞的形态学改变。Western blot法和细胞免疫荧光法检测Klf4、上皮标志分子E-cadherin、ZO-1及间质标志分子N-cadherin的表达变化。结果:在TGFβ1的诱导作用下,Hela细胞呈现出梭形、类成纤维细胞的形态,发生了上皮间质转化。Western blot和细胞免疫荧光检测结果显示,Hela细胞在TGFβ1诱导作用下,Klf4、上皮标志分子E-cadherin、ZO-1表达减少,间质标志分子N-cadherin表达增多。转染了si Klf4的Hela细胞经TGFβ1诱导后,与转染了阴性对照siRNA的细胞相比,上皮标志分子E-cadherin、ZO-1表达略有减少,而间质标志分子N-cadherin的表达则明显增多。结论:抑制Klf4表达可以促进Hela细胞的上皮间质转化。
Objective: To investigate the effect of Klf4 expression on the epithelial-mesenchymal transition in Hela cells. Methods: siRNA targeting Klf4 and negative control siRNA were designed and synthesized and transfected into Hela cells. Transfection of Hela cells with DMEM medium containing 10ng / ml TGF-β1 and 10% fetal bovine serum resulted in epithelial-mesenchymal transition. The control group was treated with TGF-β1-free medium. Morphological changes of the recorded cells were observed under an inverted microscope. Western blot and immunofluorescence were used to detect the expression of Klf4, E-cadherin, ZO-1 and N-cadherin. Results: Under the induction of TGFβ1, Hela cells showed spindle-like and fibroblast-like morphology with epithelial-mesenchymal transition. Western blot and immunofluorescence assay showed that the expression of Klf4, E-cadherin and ZO-1 in HeLa cells decreased and the expression of N-cadherin increased in Hela cells induced by TGFβ1. Transfection of si Klf4 Hela cells induced by TGFβ1, compared with the negative control siRNA transfected cells, epithelial markers E-cadherin, ZO-1 expression decreased slightly, and interstitial markers N-cadherin expression Then significantly increased. Conclusion: Inhibition of Klf4 expression can promote epithelial-mesenchymal transition in Hela cells.