Objective To investigate the effects of VP3 gene mediated by ultrasound targeted microbubble destruction (UTMD) and PEI-Gal on the apoptosis of HepG2 cells. Methods The cultured HepG2 cells were divided into 4 groups:control group, PEI-Gal group, PEI-Gal and ultrasound group, PEI-Gal plus ultrasound and microbubble group. The expression of pEGFP-VP3 in HepG2 cells 24 hours after transfection was assessed by fluorescent microscopy and flow cytometry. The expressions of VP3 mRNA and VP3 protein were measured by RT-PCR or Western Blot, and the apoptosis rate of HepG2 cells was measured by AnnexinV-FITC/Pl method. Results The transfection rate was (28.83 ± 2.07)%, the expression of VP3 mRNA and VP3 protein were 0.92 ± 0.02 and 1.65 ± 0.06, the apoptosis rate of HepG2 cells was (7.40 ± 2.12)% in PEI-Gal plus ultrasound and microbubble group, and were significantly higher than those in the other three groups (P < 0.01). Conclusions UTMD combined with PEI-Gal can increase the efficiency of VP3 gene transfecting HepG2 cells and enhance the apoptosis of HepG2 cells.%目的:探讨超声靶向微泡破碎(ultrasound targeted microbubble destruction,UTMD)联合半乳糖聚乙烯亚胺(PEI-Gal)介导凋亡素基因(VP3)诱导肝癌细胞HepG2凋亡的影响.方法:体外培养HepG2细胞,随机分为4组:(1)对照组;(2)PEI-Gal组;(3)PEI-Gal+超声组;(4)PEI-Gal+超声+微泡组.转染24 h后荧光显微镜观察pEGFP-VP3在HepG2细胞中的表达,流式细胞仪检测转染效率,RT-PcR和Western blot分别检测VP3 mRNA和蛋白表达水平,AnnexinV-FITC/PI检测细胞凋亡率.结果:流式细胞仪、RT-PCR和Western blot分析表明,PEI-Gal+超声+微泡组的转染效率为(28.83±2.07)%、VP3 mRNA水平为0.92±0.02,蛋白水平为1.65±0.06,HepG2凋亡率为(37.40±2.12)%,均显著高于其他各组(均P<0.01).结论:UTMD联合PEI-Gal可明显提高VP3基因转染HepG2细胞的效率及在HepG2细胞内的表达,增加HepG2细胞的凋亡率.
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