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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 mfn2 gene on the proliferation and chemosensitivity of hepatoma cells. Methods The experiment was divided into three groups: HepG2 cells transfected with recombinant plasmid pEGFPmfn2 as experimental group, HepG2 cells transfected with empty plasmid pEGFP? N2 as negative control group, and HepG2 cells as blank control group. The expression of mfn2mR-NA and protein in each group were detected by RT-PCR and Western Blot. The effect of mfn2 gene on the proliferation of hepatoma cells was detected by cytometry and MTT assay. The distribution of cell cycle after transfection was detected by flow cytometry. The effect of mfn2 gene transfection on chemosensitivity was detected by MTT assay. Results After 48 hours of transfection, the cell growth in mfn2 group was significantly inhibited (P <0.05), and was significantly lower than that in blank control group and transfected empty vector group (83.2 ± 1.5)%, which was significantly higher than the proportion of G0 / G1 phase in transfected empty plasmid pEGFP group and blank control group, the difference was significant (P <0.05). Experimental group HepG2 cells on the chemotherapy drug 5-fluorouracil sensitivity increased. Conclusion mfn2 can inhibit the proliferation of hepatoma cells. The mechanism may be related to the cell cycle arrest. Mfn2 can enhance the sensitivity of chemotherapeutic drugs.