外源性FHIT基因对多柔比星诱导胃癌MGC-803细胞凋亡的影响  

Influence of exogenous fragile histidine triad gene on Doxorubicin-induced apoptosis of gastric cancer cell MGC-803

作　　者：薛淑芳[1] 许洪伟[1] 张春清[1] 郝菁华[1] 秦成勇[1] 王潍博[2] 

机构地区：[1]山东大学山东省立医院消化内科,济南250021 [2]山东省立医院化疗科,济南250021

出　　处：《中国肿瘤生物治疗杂志》2007年第3期279-283,共5页Chinese Journal of Cancer Biotherapy

基　　金：山东省自然科学基金资助项目(No2004ZX05)~~

摘　　要：目的:将脆性组氨酸三联体(fragilehistidinetriad,FHIT)基因导入该基因表达缺失的人胃癌细胞株MGC-803,探讨FHIT基因表达对多柔比星诱导胃癌细胞凋亡的影响。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表达缺失的胃癌细胞MGC-803,筛选阳性克隆,同时以空载体pRcCMV转染的胃癌细胞及胃癌细胞株作为对照;以多柔比星作用于3组细胞,用MTT法测定细胞增殖的抑制率,倒置显微镜下观察细胞形态变化,用丫啶橙荧光染色与流式细胞术检测多柔比星处理前后各组胃癌细胞的凋亡率,流式细胞术检测细胞周期的变化。结果:通过流式细胞术检测,经多柔比星处理后,转染FHIT基因的MGC-803细胞凋亡水平(40.66%)与空质粒转染细胞(13.94%)及胃癌细胞(15.81%)相比明显增高,存在显著性差异(P<0.01),FHIT基因与多柔比星有轻度的协同促凋亡作用(P<0.05);同时FHIT基因转染后的胃癌细胞生长周期出现了明显的G0/G1期阻滞(74.43%vs56.30%、52.30%);丫啶橙染色亦见转染FHIT基因的胃癌细胞凋亡数明显增多,且细胞的增殖抑制率存在浓度和时间依赖性。结论:外源性FHIT基因表达与多柔比星协同促进胃癌MGC-803细胞凋亡,FHIT基因可提高胃癌细胞对多柔比星的敏感性。Objective： To study the influence of exogenous fragile histidine triad （FHIT） gene on Doxorubicin-induced apoptosis of gastric cancer cell MGC - 803. Methods： Recombinant plasmid pRcCMV-FHIT was transfected into gastric cancer cell line MGC-803 with the help of liposome and the positive clones were selected. MGC-803 cells transfected with empty pRcCMV and untransfected MGC-803 cells were taken as control and blank. After treatment with doxorubicin, the inhibition rate of the cells in 3 group was determined by MTr assay. The morphological changes of cancer cells were observed under inverted microscope and cell apoptosis was analyzed by acridine orange fluorescence staining method and flow cytometry （FCM）. Cell cycle was analyzed by FCM. Results：Stable expression of FHIT protein was obtained in transfected MGC-803 cells. FCM showed that, after treatment with doxorubicin, the apoptosis cells were significantly higher in FHIT-transfected MGC-803 cells than those in control cells and blank cells （40.66% vs 13.94% and 15.81%, P〈 0. 01 ）. There was a slight synergistic effect of FHIT gene and doxorubicin in inducing apoptosis （ P 〈 0.05 ）. We also found that cells had a G0/G1 phase arrest in FHIT-transfected cells, significantly higher than those in control cells and blank cells （74. 43% vs 56.30% and 52.30% ）. Acridine orange staining showed obvious apoptosis of cells after transfected with FHIT; the inhibition of cells was in a concentration- and time-dependent manner. Conclusion： Doxorubicin and FHIT gene can synergistically induce apoptosis of gastric cancer cell line MGC-803. FHIT gene might increase the sensitivity of gastric cancer cells to doxorubicin.

关 键 词：胃癌 脆性组氨酸三联体基因 多柔比星 基因转染 细胞凋亡 

分 类 号：R730.54[医药卫生—肿瘤]