摘要
目的探讨miR-454-3p在肝癌细胞增殖和侵袭中的作用及其可能分子学机制。方法应用实时定量聚合酶链反应技术检测肝癌组织和细胞中miR-454-3p的表达水平。体外通过CCK-8法和Matrigel-Transwell实验评估miR-454-3p人肝癌细胞增殖和侵袭能力的影响。双荧光素酶报告基因验证miR-454-3p对下游靶基因VGLL4蛋白的调控作用。通过蛋白质印迹法来检测VGLL4蛋白在人肝癌细胞中的表达水平。结果在本研究中,实验数据证明miR-454-3p在肝癌组织和细胞系中表达上调。细胞功能学实验证实过表达miR-454-3p增强肝癌细胞的增殖和侵袭能力,而抑制miR-454-3p表达则降低肝癌细胞的增殖和侵袭能力。生物信息学预测、蛋白质印迹法、qRT-PCR和荧光素酶报告分析证实VGLL4是miR-454-3p的直接靶基因,并证实增强VGLL4表达可削弱miR-454-3p促进细胞增殖和侵袭的作用。结论miR-454-3p可通过靶向抑制VGLL4基因的表达促进肝癌细胞的增殖和侵袭。
Objective To investigate the expression of miR-454-3pin hepatocellular carcinoma(HCC)and explore its effects on cell proliferation and invasion,and the target genes of HCC cells.Methods Real-time PCR was performed to detect miRNA-454-3plevels in HCC tissues and cell lines.The malignant behavior of cell lines was assessed with MTT and Transwell assays.A dual-luciferase reporter assay was used to evaluate the interaction between miR-454-3p and VGLL4.Protein expression was observed by western blotting.Results In the present study,found that miR-454-3p was up-regulated in HCC tissues and cell lines.And also demonstrated that miR-454-3p overexpression enhanced the ability of proliferation and invasion in HCC cell lines,and conversely,silencing miR-454-3p expression induced the opposite effects.VGLL4,as the direct target of miR-454-3p,was predicted by bioinformatic websites and confirmed by western blot,qRT-PCR,and luciferase reporter assay.Enforced VGLL4 expression rescued the effects of miR-454-3p mimics on cell proliferation and invasion.Conclusion The findings suggested that the miR-454-3p promoted HCC cell proliferation and invasion via targeting VGLL4.
作者
陈建清
陶元平
杨远
胡良凯
Chen Jianqing;Tao Yuanping;Yang Yuan;Hu Liangkai(The First Department of General Surgeny,Shidong Hospital,Yangpu District,Shanghai,Anhui Medical University,999 Shiguang Road,Shanghai 200438,China;The Third Department of Hepatic Surgery,Eastern Hepatobiliary Surgery Hospital,Second Military Medical University,225 Changhai Road,Shanghai 200433,China)
出处
《贵州医药》
CAS
2020年第11期1683-1686,F0003,共5页
Guizhou Medical Journal
基金
国家自然基金面上项目(No:81471038)
上海市卫生和计划生育委员会科研课题青年项目(编号:20154Y0083)
上海市卫计委青年基金(编号:20164Y0189)
国自然创新团体(编号:81521091)
国家人类资源平台(编号:2005DKA21300)。
