miR-142-5p靶向转录因子YY1调控非小细胞肺癌上皮间质转化

miR-142-5p targeting YY1 regulating epithelial-mesenchymal transition in non-small cell lung cancer

目的:探究miR-142-5p靶向转录因子YY1调控上皮间质转化(epithelial-mesenchymal transition,EMT)抑制非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞体外转移的机制。方法:qRT-PCR检测肺癌细胞系中YY1和miR-142-5p表达水平;利用生物信息学软件预测并通过双荧光素酶报告基因实验验证miR-142-5p与YY1的结合位点;Transwell实验和划痕愈合实验分析YY1和miR-142-5p过表达对肺癌细胞迁移、侵袭的影响;Western blot与qRT-PCR检测各组细胞内EMT相关蛋白的表达变化。结果:YY1在不同肺癌细胞系中均表达上调,miR-142-5p表现为表达下调;双荧光素酶报告基因实验结果进一步验证了YY1与miR-142-5p的靶向结合相关性;抑制miR-142-5p表达水平能显著提高细胞中YY1表达,而miR-142-5p过表达能显著抑制肺癌细胞中YY1的表达水平,两者表达水平呈负相关;Transwell和划痕实验表明,YY1过表达能够促进肺癌细胞转移,而miR-142-5p过表达会抑制肺癌细胞转移;YY1过表达促进EMT相关蛋白的表达,而miR-142-5p作用相反。结论:miR-142-5p负向调控YY1作用于EMT相关分子表达,进而在体外抑制NSCLC细胞的转移。

Objective:To explore the mechanism by which miR-142-5p targets YY1 to regulate epithelial-mesenchymal transition and inhibit the metastasis of non-small cell lung cancer cells in vitro and in vivo.Methods:qRT-PCR was used to detect the expression levels of YY1 and miR-142-5p in lung cancer cell lines.The dual-luciferase reporter gene assay was used to verify the targeted binding of miR-142-5p to YY1.The effect of YY1 and miR-142-5p on the cell migration and invasion by Transwell and wound-healing assay.Finally,the EMT-related proteins in each group were detected by Western blot and qRT-PCR.Results:The expression of YY1 was up-regulated and the miR-142-5p was down-regulated in lung cancer cell lines.The results of the dual-luciferase reporter assay further validated the targeted binding correlation between YY1 and miR-142-5p.Inhibiting the expression of miR-142-5p can increase the expression of YY1 significantly,However,overexpression of miR-142-5p inhibit the expression of YY1 in lung cancer cells.The expressions of YY1 and miR-142-5p in different lung cancer cell lines were negatively correlated.Transwell and wound-healing assay indicated that YY1 could enhance the metastasis ability of lung cancer cells,but overexpression of miR-142-5p could inhibit the metastasis ability of lung cancer cells.Overexpression of YY1 promotes the expression of EMT-related proteins.On the contrary,miR-142-5p constrained the expression of EMT related proteins.Conclusion:miR-142-5p may regulated the expression of EMT-related proteins by targeting YY1 involved in the lung cancer metastasis.