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目的探讨微小RNA-185(micro RNA-185,miR-185)对肺鳞癌细胞增殖和侵袭能力的作用及其可能的作用机制。方法人肺鳞癌细胞系(NCI-H520、NCI-H226)以及正常肺上皮细胞(BEAS-2B)为实验用细胞;q RT-PCR检测miR-185表达量;miR-NC(对照)和miR-185 mimics(miR-185类似物)利用Turbofect进行转染;Western blot检测E2F6的蛋白表达量;双荧光素酶报告基因检测miR-185与E2F6的靶向调控关系;MTT检测细胞的增殖能力;细胞侵袭试剂盒检测细胞的侵袭能力;构建pc DNA.3.1-E2F6重组载体,并转染肺鳞癌细胞过表达E2F6。结果 miR-185表达量在肺鳞癌细胞中显著降低(P<0.01)。miR-185靶向抑制E2F6基因(P<0.05)。转染miR-185 mimics显著上调miR-185的细胞含量(P<0.01)并抑制E2F6蛋白表达(P<0.01),肺鳞癌细胞增殖能力(P<0.05或P<0.01)和侵袭能力(P<0.05)亦显著受抑。转染pc DNA.3.1-E2F6到miR-185含量升高的细胞,E2F6蛋白表达量显著升高(P<0.01)且细胞增殖(P<0.01)和侵袭能力(P<0.05)上升。结论 miR-185可以通过靶向E2F6抑制肺鳞癌细胞增殖和侵袭。
Objective To investigate the effect of microRNA-185 (miR-185) on proliferation and invasion of lung squamous cell carcinoma and its possible mechanism. Methods Human lung squamous carcinoma cell lines (NCI-H520, NCI-H226) and normal lung epithelial cells (BEAS-2B) were used as experimental cells. The expression of miR-185 was detected by q RT- -185 mimics was transfected by Turbofect; Western blot was used to detect the expression of E2F6; Dual luciferase reporter gene was used to detect the relationship between miR-185 and E2F6; MTT was used to detect the proliferation of cells; Cell invasion kit was used to detect the invasion of cells. The pcDNA3.1-E2F6 recombinant vector was constructed and transfected into lung squamous cell carcinoma cells overexpressing E2F6. Results The expression of miR-185 in lung squamous cell carcinoma was significantly lower (P <0.01). miR-185 targeted inhibition of E2F6 gene (P <0.05). MiR-185 mimics significantly up-regulated the cell content of miR-185 (P <0.01) and inhibited the expression of E2F6 protein (P <0.01) <0.05) also significantly inhibited. E2F6 protein expression was significantly increased in cells transfected with pcDNA3.1-E2F6 and miR-185 (P <0.01), and cell proliferation (P <0.01) and invasiveness (P <0.05) increased. Conclusion miR-185 can inhibit the proliferation and invasion of lung squamous cell carcinoma cells by targeting E2F6.