eEF1A1通过正向调控NOB1的表达促进肝癌细胞的侵袭和转移  

Eukaryotic translation elongation factor 1A1 positively regulates NOB1 expression to promote invasion and metastasis of hepatocellular carcinoma cells in vitro

作　　者：张文明 项明峰[3] 郑楚骞 陈磊峰 戈进 晏琛 刘秀霞[2] ZHANG Wenming;XIANG Mingfeng;ZHENG Chuqian;CHEN Leifeng;GE Jin;YAN Chen;LIU Xiuxia(Department of General Surgery;Department of Urology;Department of Rheumatology;Jiangxi Key Laboratory of Molecular Medicine,Second Affiliated Hospital of Nanchang University,Nanchang 330000,China)

机构地区：[1]南昌大学第二附属医院普外科,江西南昌330000 [2]南昌大学第二附属医院江西分子医学重点实验室,江西南昌330000 [3]南昌大学第二附属医院泌尿外科,江西南昌330000 [4]南昌大学第二附属医院风湿免疫科,江西南昌330000

出　　处：《南方医科大学学报》2018年第10期1195-1202,共8页Journal of Southern Medical University

基　　金：国家自然科学基金(81460453);江西省青年科学基金(20171ACB21063);江西省教育厅科学技术研究项目(GJJ160164;GJJ160251);江西省研究生创新专项资金项目(YC2012-B014)~~

摘　　要：目的探讨真核翻译延长因子1A1(eEF1A1)对肝癌侵袭转移的影响机制。方法采用荧光定量PCR和Western blot法检测多种肝癌细胞系和正常肝脏细胞中eEF1A1和NI1/RPN12结合蛋白1同源物(NOB1)的mRNA和蛋白表达。肝癌细胞中干扰和过表达eEF1A1后,通过Transwell侵袭实验和RTCA实验观察肝癌细胞侵袭迁移能力的变化,并观察肝癌细胞中NOB1mRNA及蛋白表达变化。在稳定干扰eEF1A1表达的HCCLM3细胞中过表达NOB1,或在过表达eEF1A1的MHCC97h细胞中干扰NOB1的表达,分析eEF1A1和NOB1蛋白表达水平和细胞侵袭迁移能力。结果肝癌细胞中eEF1A1和NOB1表达明显高于正常肝细胞,并呈正相关。肝癌细胞中干扰eEF1A1表达可明显降低肝癌细胞的侵袭迁移能力,同时NOB1的mRNA和蛋白表达也显著被降低(P均<0.01);过表达eEF1A1后明显增加肝癌细胞的侵袭迁移能力,同时也增加NOB1的mRNA和蛋白表达(P均<0.01)。在稳定干扰eEF1A1表达的HCCLM3细胞中过表达NOB1导致NOB1表达和肝癌细胞侵袭迁移能力的恢复(P均<0.01);然而在稳定过表达eEF1A1的肝癌细胞系中降低NOB1导致NOB1表达的下调和肝癌细胞侵袭迁移能力的抑制(P均<0.01)。结论肝癌细胞中eEF1A1正向调控NOB1表达,进而影响肝癌细胞的侵袭和迁移。Objective To explore the role of eukaryotic translation elongation factor 1 A1（eEF1 A1） in regulating the invasion and metastasis of hepatocellular carcinoma（HCC） cells and the possible mechanism. Methods qRT-PCR and Western blotting were used to detect the mRNA and protein expression of eEF1 A1 and NOB1 in different HCC cell lines and normal liver cells. The invasion and migration abilities of HCC cells with eEF1 A1 knockdown or overexpression were examined using Transwell chamber assay and RTCA assay, and the changes in NOB1 m RNA and protein expressions in the cells were detected. The effects of increasing NOB1 expression in HCCLM3-sheEF1 A1 cells and decreasing NOB1 expression in eEF1 A1-overexpressing MHCC97 h cells on eEF1 A1 expression and cell invasion and migration abilities were analyzed using Western blotting,Transwell chamber assay and RTCA assay. Results The expressions of eEF1 A1 and NOB1 were significantly increased in positive correlation in HCC cells as compared with normal hepatocytes. Knockdown of eEF1 A1 significantly decreased the invasion and migration of HCC cells and reduced the mRNA and protein expression of NOB1（P〈0.01）. Overexpression of eEF1 A1 significantly enhanced invasion and migration of HCC cells and increased NOB1 mRNA and protein expressions（P〈0.01）. Increasing NOB1 expression in HCCLM3-sheEF1 A1 cells led to the restoration of NOB1 expression and cell invasion and migration abilities（P〈0.01）, whereas decreasing NOB1 in MHCC97 h-eEF1 A1 cells resulted in inhibition of NOB1 expression and cell invasion and migration（P〈0.01）. Conclusion eEF1 A1 positively regulates the expression of NOB1 to promote the invasion and migration of HCC cells in vitro.

关 键 词：肝细胞性肝癌 eEF1A1 NOB1 侵袭和迁移 

分 类 号：R735.7[医药卫生—肿瘤]