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目的:探讨miR-205-5p对Hep G2.2.15细胞增殖及HBx基因表达的影响。方法:将miR-205-5p mimics用LipofectamineTM 2000转染HepG2.2.15细胞,荧光定量PCR检测转染后细胞中miR-205-5p与HBx、E2F1 mRNA相对表达量,ELISA法检测HBxAg蛋白表达量,CCK-8法检测细胞增殖能力,流式细胞术检测细胞周期变化,Western blot检测E2F1蛋白表达。结果:miR-205-5p mimics转染Hep G2.2.15细胞后,细胞增殖能力被抑制(P<0.05),G0/G1期细胞增加、S期细胞减少(P<0.05),细胞中HBx mRNA及上清液中HBx Ag含量下降,E2F1mRNA及蛋白表达降低(P<0.05)。结论:miR-205-5p可能通过下调E2F1的表达来抑制HepG2.2.15细胞增殖,并可抑制HBx基因表达。
Objective: To investigate the effect of miR-205-5p on proliferation and HBx gene expression in Hep G2.2.15 cells. Methods: HepG2.2.15 cells were transfected with miR-205-5p mimics by LipofectamineTM 2000. The relative expression of miR-205-5p, HBx and E2F1 mRNA was detected by fluorescence quantitative PCR. The expression of HBxAg protein was detected by ELISA. The cell proliferation was detected by CCK-8 assay. The cell cycle was detected by flow cytometry. The expression of E2F1 protein was detected by Western blot. Results: The proliferation of Hep G2.2.15 cells was inhibited by miR-205-5p mimics (P <0.05), the cells in G0 / G1 phase were increased, while the cells in S phase were decreased (P <0.05) The content of HBxAg in the supernatant decreased and the expression of E2F1 mRNA and protein decreased (P <0.05). Conclusion: miR-205-5p may inhibit the proliferation of HepG2.2.15 cells and down-regulate the expression of HBx gene by down-regulating the expression of E2F1.