miR-195调控Smad7对宫颈癌细胞增殖的影响  

MiR-195 Regulates Proliferation of Cervical Cancer Cells by Targeting Smad 7

作　　者：李蓁[1] 张帆[1] 卢玉兰[1] 蔡红兵[1] LI Zhen;ZHANG Fan;LU Yulan;CAI Hongbing(Dept. of Gynecology Oncology, Zhongnan Hospital of Wuhan Universit;Hubei Clinical Cancer Study Cente;Hubei Key Laboratory of Tumor Biological Behaviors, Wuhan 430071 , China)

机构地区：[1]武汉大学中南医院妇瘤科/湖北省肿瘤医学临床研究中心/肿瘤生物学行为湖北省重点实验室

出　　处：《武汉大学学报（医学版）》2018年第4期517-522,共6页Medical Journal of Wuhan University

基　　金：国家自然科学基金青年项目(编号:81302274);国家自然科学基金面上项目(编号:81272866)

摘　　要：目的：研究miR-195对宫颈癌细胞中Smad7表达的调控和影响癌细胞增殖的作用机制。方法：通过检测miR-195在宫颈组织中的表达水平,结合临床病理资料,研究miR-195的表达与相关临床病理资料的关系。利用CCK-8检测miR-195对宫颈癌细胞增殖活性的影响。通过GFP报告基因系统,观察miR-195对Smad7结合位点蛋白表达的调控作用。用Western Blot检测miR-195对内源性Smad7及CDC25A磷酸化的影响。结果：miR-195在宫颈癌组织低表达并与临床分期及淋巴结转移密切相关（P〈0.05）。抑制miR-195表达可提高宫颈癌细胞增殖活性,差异有统计学意义（P〈0.05）。GFP报告基因系统明确Smad7为miR-195的靶基因。过表达miR-195下调Smad7蛋白水平,最终导致CDC25A的磷酸化明显下降。结论：宫颈癌细胞中miR-195可通过其靶向基因Smad7表达水平,影响TGF-β信号通路,改变CDC25A磷酸化水平,影响癌细胞的增殖。Objective：To study the mechanism of miR-195 inhibiting the proliferation of cervical cancer cells by regulating Smad7.Methods：miR-195 was quantified from cervical cancer tissues and adjacent non-tumor cervical tissues by using the RT-PCR.Correlation analyses were carried out between miR-195 expression and clinicopathological features in cervical cancer patients.CCK-8 method was used to detect the effect of miR-195 on the proliferation of SiHa and C33 Acells.The regulatory role of miR-195 on Smad7 binding site protein was observed by GFP reporter gene system.After treated with miR-195 analogue or inhibitor,the expression level of endogenous Smad7 and the phosphorylation of CDC25 A were detected by Western Blot.Results：The level of miR-195 was significantly down-regulation compared to paracarcinomatous cervical tissues（P〈0.05）,miR-195 expression was also negatively correlated with FIGO stage and venous invasion（P〈0.05）.Cell proliferation was statistically significant induced by down-regulated expression of miR-195 in SiHa and C33A cells（P〈0.05）.The GFP reporter gene system confirmed that Smad7 was predict target gene of miR-195.The over-expression of miR-195 could significant de-crease endogenous Smad7 expression,and finally the level of CDC25 Aphosphorylation also was down regulated obviously.Conclusion：In cervical cancer cells,miR-195 can affect the TGF-beta signaling pathway through its target gene Smad7 expression level,change the phosphorylation level of CDC25A,and promote the proliferation of cancer cells.

关 键 词：miRNA-195 宫颈癌组织 增殖 SMAD7 CDC25A 

分 类 号：R737.33[医药卫生—肿瘤]