逆转录病毒载体介导的端粒酶抑制基因促进肝癌细胞凋亡

Apoptosis of BEL-7402 cell line mediated by retroviral vector containing antisense human telomerase RNA

目的:探讨逆转录病毒载体介导的反义端粒酶抑制基因对肝癌细胞系BEL-7402的凋亡诱导作用.方法:用电穿孔的方法把正、反义端粒酶抑制基因导入包装细胞,包装出完整的病毒,用此病毒感染肝癌细胞系BEL-7402,绘制细胞生长曲线来研究其抗肿瘤疗效.应用MTT、流式细胞术研究细胞凋亡情况.结果:肿瘤细胞感染逆转录病毒后,肿瘤生长受到明显抑制,转染反义hTR病毒组BEL-7402细胞凋亡率显著高于转染正义hTR病毒和生理盐水组(61.32%±2.24%vs23.02%±2.13%,4.11%±1.00%,P<0.01),转染反义hTR与5-FU联用细胞凋亡率更高,为71.71%±2.53%.结论:反义端粒酶抑制基因能明显抑制肿瘤细胞的体外生长,与抗肿瘤药物5-FU有协同作用,联合用药可以增强疗效,具有明显抗肿瘤作用.

AIM： To investigate the apoptosis-inducing effect of retroviral vector containing antisense human telomerase RNA （hTR） on hepatocellular carcinoma （HCC） cell line BEL-7402. METHODS： The complete virus was obtained by importing the sense and antisense hTR into the PT67 cell line with electroporation. Bel-7402 cell line was then transfected with the virus. The therapeutic effect was evaluated by cell growth curve. MTr assay and flow cytometry were used to investigate the apoptosis of BEL-7402 cells. RESULTS： After transfection with retroviral vector, the growth of BEL-7402 cells was obviously inhibited. The apoptotic rate was significantly higher in the cells transfected with antisense hTR than that in the cells transfected with sense hTR and normal saline controls （61.32% ± 2.24% vs 23.02% ± 2.13%, 4.11% ± 1.00%, P 〈 0.01）, and the apoptotic rate was more significant when the antisense hTR-transfected cells were treated with 5-FU （71.71% ± 2.53%）. CONCLUSION： hTR can induce significant apoptosis of BEL-7402 cell line, and it plays a synergic role with 5-FU.