摘要
目的通过生物信息学技术、分子生物学等研究ENAH基因在肝癌中的表达及对肝癌细胞侵袭、运动的影响。方法检索Oncomine、TCGA、Human Protein Atlas等肿瘤数据库,分析ENAH在肝癌中的表达,免疫组化分析ENAH在肝癌组织中的表达定位及丰度,研究ENAH的调控microRNA,并通过免疫荧光、Western blot法分析miR-497-5p及ENAH对肝癌细胞侵袭、运动的影响。结果Oncomine数据库各肿瘤中ENAH差异分析,且通过Meta分析证实ENAH蛋白在肝癌中呈显著高表达。在Human Protein Atlas免疫组化数据分析肝癌组织中ENAH蛋白呈明显深染,显著高于对照组。通过TCGA数据,生存分析表明高表达的ENAH肝癌患者预后明显差于低表达组。在肝癌细胞HepG2中,miR-497-5p能够下调ENAH的表达,并且抑制HepG2细胞的paxillin磷酸化,抑制paxillin介导的细胞侵袭、极化。结论ENAH蛋白在肝癌中呈现高表达,而且ENAH的调控者miR-497-5p通过下调ENAH来抑制肝癌细胞的细胞骨架结构极化、细胞侵袭及运动。
Objective To explore the expression of ENAH in liver cancer and its effort of promoting invasion and motility of liver cancer cells.Methods The Oncomine,TCGA and Human protein atlas tumor databases were implied to analyze the expression of ENAH in hepatocellular carcinoma.The abundance of ENAH protein in hepatocarcinoma were analyzed by immunohistochemistry.The regulation of microRNA by ENAH was studied by RT-PCR and Western blot.The effects of miR-497-5p and ENAH on the invasion and movement of hepatoma cells were analyzed by immunofluorescent analysis.Results The high expression of ENAH in liver cancer was confirmed by Meta-analysis.ENAH protein was significantly stained in liver tissue at Human protein atlas immunohistochemical data.Survival analysis of ENAH in liver cancer patients by TCGA data showed that worse prognosis occurred in the low ENAH expression group.In hepatoma cell HepG2,miR-497-5p down-regulates ENAH expression,inhibits paxillin phosphorylation and suppresses the paxillin-mediated cell polarization and invasion.Conclusion ENAH protein highly expressed in hepatocarcinoma,and the miR-497-5p-ENAH participated in regulating cell cytoskeletal polarization,cell motality and invasion.
作者
慕刚刚
朱益洁
于红刚
李红艳
Mu Ganggang;Zhu Yijie;Yu Honggang(Gastroenterology Department,Renmin Hospital of Wuhan University,Hubei 430061,China)
出处
《医学研究杂志》
2020年第5期20-25,30,共7页
Journal of Medical Research
基金
国家自然科学基金青年科学基金资助项目(81602116)。
