siRNA沉默KLF4表达促进HeLa细胞上皮间质化

发布时间：2018-05-16 00:12

本文选题：Kruppel样因子 + 上皮间质转化　；参考：《现代妇产科进展》2016年04期

【摘要】：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。
[Abstract]:Objective: to investigate the effect of Klf4 expression on epithelial mesenchymal transformation of Hela cells. Methods: siRNA targeting Klf4 and negative control siRNAs were designed and synthesized and transfected into Hela cells. The epithelial mesenchymal transformation of Hela cells was induced by DMEM medium containing 10ng/ml TGF- 尾 1 and 10% fetal bovine serum, while the control group was treated with the medium without TGF- 尾 1. The morphological changes of the cells were observed under inverted microscope. The expression of Klf4, E-cadherinine ZO-1 and N-cadherin were detected by Western blot and immunofluorescence. Results: under the induction of TGF 尾 1, Hela cells showed fusiform and fibroblast-like morphology. The results of epithelial interstitial transformation. Western blot and immunofluorescence assay showed that the expression of E-cadherinine ZO-1, an epithelial marker, was decreased in Hela cells induced by TGF 尾 1. The expression of interstitial marker N-cadherin was increased. After induced by TGF 尾 1, the expression of E-cadherinine ZO-1 was slightly decreased in Hela cells transfected with si Klf4, while the expression of N-cadherin in interstitial marker was significantly increased. Conclusion: inhibition of Klf4 expression can promote the epithelial interstitial transformation of Hela cells.
【作者单位】：第四军医大学西京医院妇产科;
【基金】：国家自然科学基金(No:81301921) 陕西省自然科学基金(No:2013JM4023)
【分类号】：R329.2

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