线粒体融合素基因2对肝癌细胞的增殖及对化疗敏感性的影响

Effect of mitofusin-2 gene transfection on the proliferation and chemosensitivity of hepatocellular carcinoma cells

目的探讨线粒体融合素基因2(mfn2)对肝癌细胞的增殖及对化疗敏感性的影响。方法实验分3组:重组质粒pEGFPmfn2转染HepG2细胞为实验组,空质粒pEGFP?N2转染HepG2细胞为阴性对照组,HepG2细胞为空白对照组。RT-PCR和Western Blot分别检测转染后各组细胞mfn2mR-NA和蛋白水平的表达。采用细胞计数法和MTT法检测mfn2基因对肝癌细胞增殖的影响;流式细胞术检测转染后细胞周期的分布情况;MTT法检测转染mfn2基因对化疗敏感性的影响。结果转染48h后,转染mfn2组细胞生长开始受到明显抑制,明显低于空白对照组和转染空载体组,差异均有显著性(P<0.05);转染pEGFPmfn2组G0/G1期所占比例为(83.2±1.5)%,明显高于转染空质粒pEGFP组与空白对照组G0/G1期所占比例,差异均有显著性(P<0.05)。实验组HepG2细胞对化疗药物5-氟尿嘧啶的敏感性增强。结论mfn2对肝癌细胞具有增殖抑制作用,其机制可能与细胞周期阻滞有关;mfn2可增强化疗药物的敏感性。

Objective To investigate the effect of mitofusin-2 gene （mfn2） transfection on the proliferation and chemosensitivity of hepatocellular carcinoma cell line HepG2. Methods The experiment were divded into three groups： pEGFPmfn2 transfected cells, pEGFP-N2 transfected cells and HepG2 cells. The mRNA expression in cells was detected by RT-PCR and protein expression by Western-blot assay. Cell counting method and MTT assay were used to detect the cell proliferation of HepG2 cells. Cell cycle of HepG2 cells was observed by flow cytometry, and chemosensitivity observed by MTT assay. Results The viable count of pEGFPmfn2-transfected cells was less than that of pEGFP-transfected cells and untransfected cells at 48 h after transfection （P 〈 0. 05 ）. Flow cytometry assay showed that in the cell cycle, the G0/G1 phase proportion was significantly higher in pEGFPmfn2-transfected cells [ （83. 2 ± 1. 5 ）% ] than in pEGFP-transfected cells and untransfected cells after transfection （ P 〈 0.05 ） , and MTT assay showed that mfn2 gene could increase their ehemosensitivity to 5-FU. Conclusions Mfn2 could inhibit the proliferation of HepG2 cells, which might result from blockage of the cell cycle. It also could increase the chemosensitivity of HepG2 cells.