ACTN4通过靶向NDUFV1对食管鳞状细胞癌的细胞增殖的影响

Effect of ACTN4 on cell proliferation of esophageal squamous cell carcinoma by targeting NDUFV1

目的探究α-辅肌动蛋白-4(ACTN4)在食管鳞状细胞癌(ESCC)中的表达情况及其对ESCC细胞增殖的影响。方法采用免疫组化实验检测ESCC组织及配对正常组织中ACTN4的表达,统计学方法分析ACTN4与临床病理特征的相关性;构建ACTN4 shRNA慢病毒表达载体,利用慢病毒包装技术建立ACTN4稳定低表达的ESCC细胞株,Western blot检测敲低效果,克隆形成实验检测ACTN4敲低对细胞增殖的影响;基于前期黑色素瘤A375细胞中ACTN4敲低的蛋白质组学分析结果,选取候选下游靶蛋白并在ESCC细胞中进行验证。结果免疫组化实验结果显示,ESCC组织中的ACTN4表达量高于正常组织(P<0.0001);成功构建ACTN4 shRNA慢病毒质粒并获得稳定低表达ACTN4的ECA109细胞株;克隆形成实验结果表明,ACTN4敲低抑制ECA109细胞的增殖;ESCC细胞中ACTN4的敲低下调NADH∶泛素氧化还原酶核心亚基V1(NDUFV1)蛋白的表达。结论ESCC细胞中,高表达的ACTN4能够促进细胞增殖,并上调NDUFV1的蛋白表达。

Objective To investigate the expression of alpha-actinin-4(ACTN4)and the effect on cell proliferation in esophageal squamous cell carcinoma(ESCC).Methods The expression of ACTN4 in ESCC tissues and paired normal tissues was detected by immunohistochemistry,and the correlation between ACTN4 and the clinicopathologi-cal features was analyzed statistically.The ACTN4 shRNA lentiviral plasmids were constructed,and the stable ECA109 strain with ACTN4 shRNA knockdown was established using lentivirus packaging technology.The knockdown efficiency on protein level was checked by Western blot,and cell proliferation was detected by colony formation assays.The downstream target proteins were validated in ESCC cell line ECA109 based on the previous proteomics analyses in melanoma cell line A375 with or without ACTN4 shRNA knockdown.Results The expression of ACTN4 in ESCC tissues was significantly higher than that of normal tissues.ACTN4 shRNA stable knockdown ECA109 cell strains were successfully constructed.The results of colony formation assays showed that ACTN4 knockdown inhibited the cell proliferation and down-regulated NADH∶Ubiquinone oxidoreductase core subunit V1(NDUFV1)protein expression in ECA109 cells.Conclusion Upregulation of ACTN4 in ESCC cells promotes the cell proliferation and enhances the protein expression of NDUFV1.