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目的判断miR-762在卵巢癌中的作用,以及miR-762对卵巢癌细胞增殖的影响。方法通过实时荧光定量聚合酶链反应(quantitative polymerase chain reaction,qPCR)分析40位卵巢癌患者标本中miR-762和menin蛋白的表达情况,进一步通过荧光素酶报告基因实验分析miR-762对menin蛋白的调节作用。在人卵巢上皮性SKOV3细胞中分别过表达或沉默miR-762后,通过四甲基偶氮唑盐(methylthiazolyldiphenyl-tetrazolium bromide,MTT)实验观察miR-762对细胞增殖的影响,进一步通过蛋白免疫印迹(western blot)实验检测miR-762对menin蛋白及Wnt通路中增殖相关蛋白的调节作用。结果 qPCR检测发现,与癌旁组织相比miR-762在肿瘤中表达较高,western blot和qPCR实验发现,menin蛋白在卵巢癌中表达较低。报告基因实验证明,miR-762可以直接打靶menin蛋白。MTT检测发现miR-762过表达后可以通过抑制menin蛋白及激活Wnt通路来促进卵巢癌细胞系SKOV3细胞的增殖。结论 miR-762在人卵巢癌组织中呈高表达,并可能通过抑制menin蛋白及激活Wnt通路从而促进卵巢癌的进程。
Objective To determine the role of miR-762 in ovarian cancer and the effect of miR-762 on the proliferation of ovarian cancer cells. Methods The expression of miR-762 and menin in 40 specimens of ovarian cancer was analyzed by quantitative polymerase chain reaction (qPCR). The luciferase reporter assay was used to analyze the effect of miR-762 on the expression of menin protein The regulatory role. After overexpression or silencing of miR-762 in human ovarian epithelial SKOV3 cells, the effect of miR-762 on cell proliferation was observed by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. Western blot (western blot) was used to detect the regulatory effect of miR-762 on the expression of menin protein and Wnt signaling. Results qPCR showed that miR-762 was highly expressed in tumor tissues compared with adjacent tissues. Western blot and qPCR showed that menin protein was low in ovarian cancer. Reporter gene experiments show that miR-762 can directly target menin protein. The results of MTT assay showed that miR-762 overexpression promoted the proliferation of ovarian cancer cell line SKOV3 by inhibiting menin protein and activating Wnt pathway. Conclusion miR-762 is overexpressed in human ovarian cancer tissues and may promote ovarian cancer progression by inhibiting the expression of menin protein and activating Wnt pathway.