microRNA-320d通过PBX3抑制子宫内膜癌JEC细胞上皮-间质转化功能  

MicroRNA-320d Inhibits Epithelial Mesenchymal Transition Function in Endometrial Carcinoma JEC Cells by PBX3

作　　者：王晶[1] 龚凤球[1] 何科[1] 姚书忠[1] 牛刚[1] WANG Jing GONG Feng-qiu HE Ke YAO Shu-zhong NIU Gang(Department of Obstetrics and Gynecology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China)

机构地区：[1]中山大学附属第一医院妇产科,广东广州510080

出　　处：《中山大学学报（医学科学版）》2017年第5期651-657,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学青年基金(81602723);广东省科技发展专项(公益研究与能力建设;2016A020215214);广东省医学科研基金(A2015130)

摘　　要：【目的】研究microRNA-320d(miR-320d)对子宫内膜癌JEC细胞上皮-间质转化功能的影响及其相关机制。【方法】JEC子宫内膜癌细胞株分别转染miR-320d mimics和negative control mimic,分别作为M320d、NCM组,并设立未转染对照control组,采用RT-PCR法检测各组细胞miR-320d含量。Transwell实验检测3组细胞迁移能力和侵袭能力。Westernblot法检测3组细胞α-Catenin和PBX3表达水平,及PBX3过表达对miR-320d抑制EMT的拮抗作用。双荧光素酶实验检测miR-320d与PBX3的关系。【结果】M320d组miR-320d的表达水平为control组的(808.25±15.58)倍(P<0.05)。M320d组迁移细胞数量29.56±0.59低于control组的94.48±1.02(P<0.05)。M320d组侵袭细胞数量7.33±0.84低于control组的86.28±3.51(P<0.05)。M320d组细胞α-Catenin、E-cadherin蛋白表达量(0.365±0.017、0.261±0.008)高于对照组(0.114±0.010、0.151±0.021),Vimentin蛋白表达量(0.105±0.009)低于对照组(0.211±0.025),PBX3蛋白表达量(0.181±0.002)低于对照组(0.311±0.007)。PBX3过表达M320d组子宫内膜癌细胞中α-Catenin、E-cadherin蛋白表达量(0.139±0.019、0.143±0.007)低于对照组(0.399±0.034、0.261±0.017),Vimentin蛋白表达量(0.234±0.008)高于对照组(0.105±0.009),差异均有统计学意义(P<0.05)。双荧光素酶检验结果显示PBX3为miR-320d的下游靶基因。【结论】miR-320d可能通过降低下游靶基因PBX3水平影响EMT相关蛋白表达,抑制子宫内膜癌JEC细胞的上皮-间质转化功能。【Objective】To investigate the inhibitory effect and mechanism of the microRNA-320d（miR-320d）on epithelial mesenchymal transition in endometrial carcinoma JEC cells. 【Methods】JEC endometrial carcinoma cell lines were transfected with miR-320 d mimics or negative control mimic,respectively,as M320 d or NCM group. Control group was established with untreated JEC endometrial carcinoma cells. miR-320 d content in each group was detected by RT-PCR method. Transwell assay was used to detect the migration and invasion ability of the 3 groups. Western-blot assay was used to detect the expressions of α-Catenin,E-cadherin,Vimentin and PBX3 protein in 3 groups. Antagonistic effect of PBX3 overexpression on miR-320 d inhibition of EMT was detected by western blot assay. The relationship between miR-320 d and PBX3 was detected by dual luciferase assay.【Results】The expression level of miR-320 d in M320 d group was significantly up-regulated,and the expression level of miR-320 d was 808.25 ± 15.58 times higher than that of control group（P〈0.05）. The number of migrating cells in M320 d group was 29.56 ± 0.59,which was significantly lower than that of control group at 94.48 ± 1.02（P〈0.05）. The number of invasive cells in M320 d group was 7.33±0.84,which was significantly lower than that of group control 86.28 ± 3.51（P〈0.05）. Compared with control group,the expression ofα-Catenin and E-cadherin protein was significantly increased,the expression of Vimentin protein was significantly decreased,and the expression of PBX3 protein was significantly decreased. After PBX3 overexpression,the expression of α-Catenin and E-cadherin protein were significantly decreased,the expression of Vimentin protein were significantly increased. Dual luciferase assay showed that PBX3 is a downstream target gene of miR-320d（P〈0.05）.【Conclusion】miR-320 d may inhibit the expression of EMT related protein through the downstream target gene PBX3 and inhibit the epithelial mesenchymal transition function of en

关 键 词：子宫内膜癌 微小RNA-320d 上皮-间质转化 

分 类 号：R711[医药卫生—妇产科学]