超声靶向微泡破碎联合半乳糖聚乙烯亚胺介导凋亡素基因诱导肝癌细胞凋亡的研究

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24h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。

Apoptosis of HepG2 cells induced by VP3 gene mediated by ultrasound targeted microbubble destruction and PEI-Gal

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.