人胃癌细胞株MGC-803 FHIT基因转染对阿霉素敏感性的影响

Effect of fragile histidine triad gene transfection into the human gastric cancer cell line MGC-803 on Adriamycin

目的:将FHIT基因导入该基因表达缺失的人胃癌细胞株MGC-803,探讨胃癌中FHIT基因表达对阿霉素(ADM)敏感性的影响。方法:将载有人外源性FHIT基因的真核表达质粒pRcCMV-FHIT用脂质体介导转入FHIT蛋白表达缺失的胃癌细胞株MGC-803,筛选阳性克隆,同时以空载体pRcCMV转染的胃癌细胞及未转染的胃癌细胞株作为对照,以ADM作用于三组细胞,用MTT法测定细胞的生长抑制率,用流式细胞仪检测ADM处理前后各组胃癌细胞的凋亡率及细胞周期的变化。结果:经ADM处理后,转染FHIT基因的MGC-803细胞的凋亡水平(40.66%)与空质粒转染组(13.94%)及胃癌细胞株组(15.81%)相比明显增高(P<0.01),并且FHIT基因与ADM有轻度的协同促进凋亡作用(P<0.05);同时ADM处理前后实验组胃癌细胞的生长周期较对照组均出现了明显的G0/G1期阻滞(74.43%vs 56.30%,99.27%vs 95.10%),且细胞的生长抑制率存在浓度和时间依赖性。结论:外源性FHIT基因表达与ADM协同促进MGC-803细胞凋亡及细胞周期阻滞,FHIT基因可增加胃癌细胞对ADM的敏感性。

Objective： The wild type FHIT gene was transfected into the human gastric cancer cell line MGC-803 in which the FHIT gene had been totally lost so as to investigate its effect on Adriamycin （ADM）. Methods： The FHIT gene was transfected into the gastric cancer cell line MGC-803 by liposome. Western bolt assay was employed to determine the expression of FHIT. Cancer cells which were transfected with empty vector pRcCMV and the non-transfected gastric cancer cell line MGC-803 were viewed as a control and a blank respectively. After treatment with Adriamycin, an MTr colormetric assay was applied to determine the inhibition ratio. The apoptosis and cell cycle were analyzed by flow cytometry （FCM）. Results： A stable expression of FHIT protein was obtained in transfected MGC-803 ceils. After treatment with Adriamycin, apoptosis cells were significantly more in FHIT-transfected MGC-803 ceils than in control cells and blank ceils （40.66% VS 13.94%, 15.81%, P 〈 0.01）. Adriamycin-induced apoptosis can be enhanced by wild FHIT expression （P 〈 0.05）. Before and after treatment of Adriamycin, the rates of G0/G1 phase were statistically higher in FHIT-transfected cells than in control cells （74.43% VS 56.30%, 99.27% VS 95.10%, respectively）. Conclusion： The FHIT gene might be involved in apoptosis and blocks the eell eyele of gastrie eaneer eells. Expression of the wild FHIT gene ean inerease the sensitivity of gastrie eaneer eell line MGC-803 to Adriamyein.