摘要
[目的]探讨miR-373-3p在肝细胞癌中的生物学作用和分子机制。[方法]用miR-373-3p模拟物或miR-373-3p抑制剂转染肝细胞癌细胞系。通过MTT、克隆形成和Transwell实验鉴定miR-373-3p对肝细胞癌细胞增殖和迁移能力的影响。Western blot和实时荧光定量逆转录聚合酶链反应(real-time quantitative reverse transcription PCR,qRT-PCR)检测相关基因蛋白和mRNA的表达水平。采用双荧光素酶法验证miR-373-3p与YWHAZ的关系。[结果]QRT-PCR检测显示,miR-373-3p在肝癌组织中的表达明显低于癌旁组织,且miR-373-3p的低表达与肝癌患者的肿瘤大小、血管浸润和不良预后显著相关。体外细胞学实验结果显示,过表达miR-373-3p能够显著抑制肝癌细胞的增殖和侵袭能力,而下调miR-373-3p的表达则作用相反。双荧光素酶报告、qRT-PCR和Western blot实验证实,miR-373-3p能够直接靶向YWHAZ的3′-非翻译区(UTR)且抑制YWHAZ在肝癌细胞中的表达。过表达的YWHAZ能够抵消miR-373-3p对肝细胞癌细胞增殖和转移的抑制作用。[结论] Mi R-373-3p可能是一种新的潜在的肝癌治疗靶点。
[Purpose]To investigate the biological function and molecular mechanism of miR-373-3p in hepatocellular carcinoma.[Methods]Hepatocellular carcinoma(HCC)cells were transfected with miR-373-3p mimics or miR-373-3p inhibitor.Proliferation and migration were identified by MTT assay,colony formation and transwell assays.Protein and mRNA expression levels of associated genes were measured by Western blot and real-time quantitative reverse transcription PCR(q RT-PCR).Dual luciferase assay was used to determine the relation of miR-373-3p to YWHAZ gene.[Results]The results of qRT-PCR assay demonstrated that expression of miR-373-3p was significantly lower in HCC tissues compared with adjacent liver tissues.The lower miR-373-3p expression significantly associated with tumor size,vascular infiltration and poor prognostic outcome in HCC patients.In vitro,ectopic overexpression of miR-373-3p significantly inhibited cell proliferation and invasion capability,while down-regulated miR-373-3p had reversed effects.Furthermore,luciferase reporter gene assay,qRT-PCR,and Western blot assays demonstrated that miR-373-3p targeted 3′-untranslated region(UTR)of YWHAZ gene and inhibited its expression in HCC cells.Overexpression of YWHAZ partially abolished the tumor suppressing effects induced by upregulating miR-373-3p in HCC cells.[Conclusion]MiR-373-3p might represent a novel potentially therapeutic target for HCC.
作者
陶元平
姚乃心
黄智平
周伟平
汪珍光
TAO Yuan-ping;YAO Nai-xin;HUANG Zhi-ping;ZHOU Wei-ping;WANG Zhen-guang(Eastern Hepatobiliary Surgery Hospital,Naval Military Medical University,Shanghai 200433,China;College of Basic Medicine,Naval Military Medical University,Shanghai 200433,China)
出处
《中国肿瘤》
CAS
CSCD
北大核心
2019年第7期549-556,共8页
China Cancer
基金
上海市卫计委青年基金(20164Y0189)
国家自然科学基金创新团体(81521091)
国家人类资源平台(2005DKA21300)
上海市卫生和计划生育委员会科研课题青年项目(20154Y0083)
