南方医科大学学报

南方医科大学学报 ›› 2018, Vol. 38 ›› Issue (10): 1195-.doi: 10.12122/j.issn.1673-4254.2018.10.07

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eEF1A1通过正向调控NOB1的表达促进肝癌细胞的侵袭和转移

张文明，项明峰，郑楚骞，陈磊峰，戈进，晏琛，刘秀霞   

  • 出版日期:2018-10-20 发布日期:2018-10-20

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

  • Online:2018-10-20 Published:2018-10-20

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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NOB1 mRNA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。

Abstract: Objective To explore the role of eukaryotic translation elongation factor 1A1 (eEF1A1) 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 eEF1A1 and NOB1 in different HCC cell lines and normal liver cells. The invasion and migration abilities of HCC cells with eEF1A1 knockdown or overexpression were examined using Transwell chamber assay and RTCA assay, and the changes in NOB1 mRNA and protein expressions in the cells were detected. The effects of increasing NOB1 expression in HCCLM3-sheEF1A1 cells and decreasing NOB1 expression in eEF1A1-overexpressing MHCC97h cells on eEF1A1 expression and cell invasion and migration abilities were analyzed using Western blotting, Transwell chamber assay and RTCA assay. Results The expressions of eEF1A1 and NOB1 were significantly increased in positive correlation in HCC cells as compared with normal hepatocytes. Knockdown of eEF1A1 significantly decreased the invasion and migration of HCC cells and reduced the mRNA and protein expression of NOB1 (P<0.01). Overexpression of eEF1A1 significantly enhanced invasion and migration of HCC cells and increased NOB1 mRNA and protein expressions (P< 0.01). Increasing NOB1 expression in HCCLM3-sheEF1A1 cells led to the restoration of NOB1 expression and cell invasion and migration abilities (P<0.01), whereas decreasing NOB1 in MHCC97h-eEF1A1 cells resulted in inhibition of NOB1 expression and cell invasion and migration (P<0.01). Conclusion eEF1A1 positively regulates the expression of NOB1 to promote the invasion and migration of HCC cells in vitro.