miR-138-5p靶向TCF4调节眼葡萄膜黑色素瘤细胞的恶性生物学行为  

MiR-138-5p targeting TCF4 regulates the malignant biological behavior of ocular uveal melanoma cells

作　　者：魏丽[1] 连红梅[2] 刘鹏[1] 刘兴华[1] 吴书一[3] 刘萌萌[1] Wei Li;Lian Hongmei;Liu Peng(Dept of Pathology,The Third People′s Hospital of Zhengzhou,Zhengzhou 450045;Dept of Anesthesia,The Third People′s Hospital Affiliated to Zhengzhou University,Zhengzhou 450045)

机构地区：[1]郑州市第三人民医院病理科,郑州450045 [2]郑州大学附属第三人民医院麻醉科,郑州450045 [3]郑州市第三人民医院中心实验室,郑州450045

出　　处：《安徽医科大学学报》2021年第6期887-893,共7页Acta Universitatis Medicinalis Anhui

基　　金：2019年度河南省医学科技攻关计划项目(编号:LHGJ20191025)。

摘　　要：目的探讨miR-138-5p与转录因子4(TCF4)对眼葡萄膜黑色素瘤MuM-2B细胞恶性生物学行为的影响。方法miR-138-5p mimic和pcDNA-TCF4分别或共转染MuM-2B细胞,构建小鼠体内移植瘤模型。RT-qPCR检测细胞转染效率;生物信息学软件预测miR-138-3p和TCF4靶向位点;双荧光素酶报告基因实验验证靶向关系;克隆形成实验检测细胞增殖;Transwell实验检测细胞侵袭能力;Western blot检测TCF4及上皮-间质转化(EMT)相关蛋白表达;免疫组化检测Ki67、VEGF、Vimentin在肿瘤组织中的表达。结果与对照组相比,转染组miR-138-5p和TCF4均升高(P<0.05),且miR-138-5p和TCF4是靶向关系。miR-138-5p过表达通过靶向TCF4抑制MuM-2B细胞增殖和侵袭力并下调E-cadherin和Vimentin蛋白表达量,上调N-cadherin蛋白表达量(P<0.05)。在体内移植瘤中,miR-138-5p过表达减轻肿瘤重量(P<0.05),miR-138-5p过表达下调Ki67、VEGF、Vimentin在肿瘤组织中阳性细胞率(P<0.05)。结论miR-138-5p过表达可下调TCF4水平,进而抑制葡萄膜黑色素瘤MuM-2B细胞增殖、侵袭及EMT。Objective To investigate the effects of miR-138-5p and TCF4 on the malignant biological behavior of MuM-2B cells in uveal melanoma.Methods MuM-2B cells were transfected or co-transfected with miR-138-5p mimic and pcDNA-TCF4,respectively.A mouse model of in vivo xenograft tumor was established.Cell transfection efficiency was detected by RT-qPCR.Bioinformatics software predicted miR-138-5p and TCF4 target sites.Dual luciferase reporter gene assay was used to verify the targeted relationship.Clone formation assay was used to detect the ability of cell clone formation.The invasion ability of cells was detected by Transwell assay.Western blot analysis of TCF4 and epidermal mesenchymal transformation(EMT)related protein expression.Immunohistochemistry was used to detect the expression of Ki67,VEGF and Vimentin in tumor tissues.Results Compared with control group,miR-138-5p and TCF4 were up-regulated in transfected group(P<0.05).Between miR-138-5p and TCF4 were target relationship.Overexpression of miR-138-5p inhibited MuM-2B cell proliferation,invasion ability as well as up-regulating the protein expressive levels of E-cadherin and Vimentin and down-regulating the protein expressive levels of N-cadherin by targeting TCF4 in Mum-2B cells(P<0.05).In transplanted tumors in vivo,overexpression of miR-138-5p significantly reduced tumor weight(P<0.05),and overexpression of miR-138-5p down-regulated the positive cell rate of Ki67,VEGF and Vimentin in tumor tissues(P<0.05).Conclusion The overexpression of miR-138-5p down-regulated the TCF4 levels,and further inhibited the cell proliferation,invasion and EMT of uveal melanin Mum-2B cells.

关 键 词：miR-138-5p 转录因子4 MuM-2B细胞 克隆形成 细胞侵袭 

分 类 号：R773.9[医药卫生—眼科]