miR-7-5p靶向调控ATG4A介导自噬对子宫内膜癌细胞增殖、侵袭的影响  

miR-7-5p regulates autophagy to inhibit proliferation and invasion of endometrial cancer cell by targeting ATG4A

作　　者：叶文蔚[1] 杨晶金[2] 应灵潇 柴芝红 蔡仙丽[1] YE Wenwei;YANG Jingjin;YING Lingxiao(Department of Gynecology,Zhejiang Taizhou Municipal Hospi-tal,Taizhou 318000,China)

机构地区：[1]台州市立医院妇科,浙江台州318000 [2]台州学院医学院

出　　处：《全科医学临床与教育》2020年第11期980-984,F0002,共6页Clinical Education of General Practice

基　　金：台州市科技计划项目(1702KY31)。

摘　　要：目的观察miR-7-5p在子宫内膜癌组织中的表达情况,并探讨miR-7-5p对子宫内膜癌细胞(HEC-1B)增殖、侵袭的影响及其机制。方法利用逆转录聚合酶链反应法(qRT-PCR)实验检测42例子宫内膜癌患者子宫内膜癌组织和癌旁正常组织中miR-7-5p的表达水平,并分析其与临床病理学特征的关系;CCK-8法及Tran⁃swell试验检测过表达miR-7-5p后HEC-1B细胞增殖及侵袭情况;通过生物信息学预测网站预测到自噬相关基因4A(ATG4A)与miR-7-5p有互补结合位点,并利用双荧光素酶报告系统鉴定它们的靶向关系;再通过Western blot实验检测上调miR-7-5p表达后HEC-1B细胞中ATG4A、LC3-Ⅱ、p62的表达情况,以及敲低ATG4A表达后LC3-Ⅱ、p62的表达情况。结果miR-7-5p在子宫内膜癌组织中相对表达量明显低于癌旁组织(t=18.65,P<0.05),其表达量与FIGO分期、肿瘤浸润深度及淋巴结转移密切相关(t分别=2.45、3.40、2.27,P均<0.05);上调miR-7-5p表达能明显抑制HEC-1B细胞的增殖能力以及侵袭能力(t分别=59.89、8.99,P均<0.05);miR-7-5p靶向调控ATG4A的表达,上调miR-7-5p水平能降低ATG4A、LC3-Ⅱ的表达(t分别=7.82、12.88,P均<0.05),升高p62的表达(t=-10.42,P<0.05),而敲低ATG4A水平后LC3-Ⅱ表达降低(t=11.41,P<0.05),p62表达升高(t=-23.13,P<0.05)。结论miR-7-5p与子宫内膜癌的发展及预后密切相关,miR-7-5p可通过靶向调控ATG4A基因的表达,抑制细胞自噬活性,从而降低HEC-1B细胞的增殖及侵袭水平。Objective To observe the expression of miR-7-5p in endometrial carcinoma and explore the effect and mechanism of miR-7-5p on the proliferation and apoptosis of endometrial carcinoma cells.Methods The expressions of miR-7-5p in 42 cases with endometrial carcinoma and normal endometrial tissues were detected by qRT-PCR,and its relationship with clinicopathological features was analyzed.CCK-8 method and Transwell test were used to detect the proliferation and invasion of HEC-1B cells after up-regulation of miR-7-5p expression.The bioinformatics prediction website predicted that ATG4A and miR-7-5p had complementary binding sites.Their targeting relationship could be identified by double luciferase reporting system.Western blot and qRT-PCR were used to detect the expression of ATG4A,LC3-Ⅱ,and p62 in HEC-1B cells after up-regulation of miR-7-5p expression,and the expression of LC3-Ⅱand p62 after down-regulation of ATG4A expression.Results The relative expression of miR-7-5p in endometrial carcinoma was significantly lower than that in normal endometrial tissues(t=18.65,P<0.05),and it was closely related to FIGO stage,tumor invasion depth and lymph node metastasis(t=2.45,3.40,2.27,P<0.05).Up-regulation of miR-7-5p expression could significantly inhibit the proliferation and invasion of endometrial carcinoma HEC-1B cells(t=59.89,8.99,P<0.05).Up-regulation of miR-7-5p expression reduced the ATG4A expression and LC3-Ⅱlevel(t=7.82,12.88,P<0.05),but increased the expression of p62(t=-10.42,P<0.05).However,after knocking down ATG4A level,the expression of LC3-Ⅱdecreased(t=11.41,P<0.05),and the expression of p62 increased(t=-23.13,P<0.05).Conclusion miR-7-5p is closely related to the development and prognosis of endometrial cancer.miR-7-5p could inhibit autophagy activity by targeting ATG4A gene expression,and thus reduce the proliferation and invasion of endometrial cancer cell.

关 键 词：miR-7-5p 自噬相关基因4A 子宫内膜癌 自噬 增殖 侵袭 

分 类 号：R737.33[医药卫生—肿瘤]