Eukaryotic translation elongation factor 1A1 promotes proliferation of hepatocellular carcinoma cells by regulating cyclin D2 expression / 肿瘤

ABSTRACT

Objective: To study the effects of eukaryotic translation elongation factor 1A1 (eEF1A1) on the cell cycle and proliferation of hepatocellular carcinoma (HCC) cells, and to explore the molecular mechanism. Methods: The expression of eEF1A1 protein in 101 HCC tissues and their adjacent tissues were detected by immunohistochemistry, and the correlations between the expression of eEF1A1 protein and the clinicopathologic parameters of HCC patients was further analyzed. The expressions of eEF1A1 mRNA and protein in HCC cells (including Huh7, SMMC7721, MHCC97H and Hep3B) and the normal liver cells HL-7702 were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The specific shRNA-1/2 targeting eEF1A1 gene (eEF1A1-shRNA-1/2) and the overexpression plasmid pcDNA3.1- eEF1A1 were respectively transfected into Huh7 and SMMC7721 cells, and the alteration of eEF1A1 expression was verified by real-time fluorescent quantitative PCR and Western blotting, respectively. Then the proliferation and cell cycle of HCC cells were detected by EdU and FCM assays, respectively. In addition, the expressions of cyclin D2 and the signal transducer and activator of transcription 1 (STAT1)-related pathway proteins in Huh7 cells transfected with eEF1A1-shRNA-1/2 and in SMMC7721 cells transfected with pcDNA3.1- eEF1A1 were detected by Western blotting. Results: The expression level of eEF1A1 protein in HCC tissues was markedly higher than that in the adjacent tissues (P 0.05) in SMMC7721 cells after eEF1A1 overexpression and treatment with STAT1 inhibitor fludarabine. Conclusion: eEF1A1 regulates the expression of cyclin D2 through STAT1 pathway, thereby promoting the cell cycle progression and proliferation of primary HCC cells.