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目的探讨Alk B同源蛋白5(Alk B homologue 5,ALKBH5)对正常肝细胞系L-02和肝癌细胞系Hep G2的增殖、周期、凋亡的影响。方法通过慢病毒稳定转染ALKBH5基因的重组载体(p EGFP-C1b-ALKBH5)至肝癌细胞系Hep G2和肝细胞系L-02中,Western印迹鉴定绿色荧光蛋白(GFP-ALKBH5)的稳定表达;实验分GFP-ALKBH5慢病毒组和GFP慢病毒组;细胞增殖实验(cell counting kit-8,CCK-8)检测两组细胞的增殖能力;流式细胞术检测两组细胞周期和细胞凋亡的情况;平板克隆形成实验检测两组细胞的生长能力。结果与GFP对照组相比,过表达GFP-ALKBH5可显著抑制Hep G2和L-02细胞的增殖,引起Hep G2和L-02细胞的周期阻滞,但不影响细胞的凋亡。结论ALKBH5可显著抑制肝癌细胞Hep G2和肝细胞L-02的增殖能力,扮演了抑癌基因的角色,可能在肝细胞癌的发生和发展中起重要作用。
Objective To investigate the effects of Alk B homologue 5 (ALKBH5) on proliferation, cell cycle and apoptosis of normal liver cell line L-02 and hepatocellular carcinoma cell line Hep G2. Methods Stable expression of green fluorescent protein (GFP-ALKBH5) was confirmed by Western blotting using recombinant lentivirus vector (p EGFP-C1b-ALKBH5) to Hep G2 and L-02 hepatocellular carcinoma cell lines. The GFP-ALKBH5 lentivirus group and GFP lentivirus group were divided into experimental group and control group. Cell proliferation kit (CCK-8) was used to detect the proliferation of the two groups. Flow cytometry was used to detect cell cycle and apoptosis Situation; plate clone formation test to test the growth ability of two groups of cells. Results Compared with the GFP control group, overexpression of GFP-ALKBH5 significantly inhibited the proliferation of Hep G2 and L-02 cells and caused cell cycle arrest in Hep G2 and L-02 cells but not apoptosis. Conclusion ALKBH5 can significantly inhibit the proliferation of Hep G2 cells and L-02 cells and play a role of tumor suppressor gene, which may play an important role in the occurrence and development of hepatocellular carcinoma.