miR-613靶向PTBP1抑制子宫内膜癌细胞侵袭和迁移的实验研究  

Experimental study of miR-613 targeting PTBP1 to inhibit the invasion and migration of endometrial cancer cells

作　　者：顾少君 钱春艳[2] 傅欢泉[1] GU Shao-jun;QIAN Chun-yan;FU Huan-quan(Department of Laboratory,the First People's Hospital of Xiaoshan District,Hangzhou,Zhejiang 311200,China;不详)

机构地区：[1]杭州市萧山区第一人民医院检验科,浙江杭州311200 [2]杭州市余杭区第一人民医院检验科,浙江杭州311100

出　　处：《中华全科医学》2022年第5期781-784,827,共5页Chinese Journal of General Practice

基　　金：浙江省基础公益研究计划项目(LGF19H200002)。

摘　　要：目的探讨微RNA-613(miR-613)靶向多聚嘧啶区结合蛋白1(PTBP1)对子宫内膜癌细胞(Ishikawa)增殖、迁移及侵袭的影响。方法采用RT-qPCR法检测子宫内膜癌组织miR-613及PTBP1 mRNA表达;体外培养Ishikawa细胞并对Ishikawa细胞进行干预,过表达miR-613、抑制PTBP1表达或共同过表达miR-613与PTBP1,分别检测Ishikawa细胞增殖、迁移与侵袭及PTBP1、基质金属蛋白酶2(MMP-2)、MMP-9蛋白表达情况;双荧光素酶报告实验验证miR-613与PTBP1的靶向关系。结果子宫内膜癌组织中miR-613表达水平为0.48±0.09,显著低于癌旁组织的1.03±0.11(P<0.05),PTBP1 mRNA表达水平为2.78±0.23,显著高于癌旁组织的1.01±0.12(P<0.05),miR-613与PTBP1 mRNA呈负相关关系(r=-0.523,P<0.05);双荧光素酶报告基因检测结果显示,miR-613靶向负调控PTBP1表达;过表达miR-613与抑制PTBP1表达,Ishikawa细胞存活率、细胞迁移与侵袭数、PTBP1、MMP-2及MMP-9表达水平显著降低(均P<0.05);过表达PTBP1逆转过表达miR-613对Ishikawa细胞增殖、迁移与侵袭的抑制作用。结论miR-613在子宫内膜癌组织中低表达,过表达miR-613可通过靶向抑制PTBP1表达,抑制人子宫内膜癌细胞的增殖、迁移及侵袭。Objective To investigate the effects of miR-613 targeting polypyrimidine tract-binding protein 1(PTBP1)on the proliferation,migration and invasion of endometrial cancer cells(Ishikawa).Methods Real-time fluorescent quantitative PCR(RT-qPCR)was conducted to detect the expression of miR-613 and PTBP1 mRNA in endometrial cancer tissues.Ishikawa cells were cultured in vitro,and intervened to overexpress miR-613,inhibit PTBP1 expression,or co-overexpress miR-613 and PTBP1.The proliferation,migration and invasion of Ishikawa cells and the protein expression levels of PTBP1,matrix metalloproteinase-2(MMP-2)and MMP-9 were detected respectively.Dual-luciferase reporter assay was conducted to verify the targeting relationship between miR-613 and PTBP1.Results The expression level of miR-613 in endometrial cancer tissue was 0.48±0.09,which was significantly lower than that in paracancerous tissue(1.03±0.11,P<0.05).The expression level of PTBP1 mRNA was 2.78±0.23,which was significantly higher than that in paracancerous tissue(1.01±0.12,P<0.05).MiR-613 was negatively correlated with PTBP1 mRNA(r=-0.523,P<0.05).The results of dual-luciferase reporter gene assay showed that miR-613 targeted and negatively regulated the expression of PTBP1.Overexpression of miR-613 and inhibition of PTBP1 expression significantly decreased the survival rate of Ishikawa cells,the number of cell migration and invasion,and the expression levels of PTBP1,MMP-2 and MMP-9(all P<0.05).Overexpression of PTBP1 reversed the inhibitory effects of overexpression of miR-613 on Ishikawa cell proliferation,migration and invasion.Conclusion The expression of miR-613 is low in endometrial cancer tissues.Overexpression of miR-613 can inhibit the proliferation,migration and invasion of human endometrial cancer cells by targeting and inhibiting the expression of PTBP1.

关 键 词：微RNA-613 多聚嘧啶区结合蛋白1 子宫内膜癌细胞 增殖 迁移 侵袭 

分 类 号：R737.33[医药卫生—肿瘤] R730.2[医药卫生—临床医学]