Bcl-XL siRNA对人胃腺癌MGC-803细胞药物敏感性的影响  

Bcl-XL siRNA sensitizes human MGC-803 gastric cancer cells to 5-fluorouracil and diallyl disulfide

作　　者：钟苗[1] 雷小勇[1] 冯兰芳[1] 朱炳阳[1] 唐圣松[1] 廖端芳[1] 

机构地区：[1]南华大学生命科学与技术学院药物药理研究所

出　　处：《中国临床药理学与治疗学》2006年第1期33-38,共6页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金项目(№30300426);湖南省教育厅青年基金(№03B034)

摘　　要：目的:研究靶向Bcl-XL基因的小干扰RNA对胃腺癌MGC-803细胞Bcl-XL基因表达的作用及对药物敏感性的影响。方法:构建的Bcl-XLsiRNA载体或阴性siRNA并稳定转染到胃癌MGC-803细胞,G418抗生素筛选阳性克隆,克隆扩大培养,免疫荧光观察细胞蛋白表达。用不同浓度的5-FU或DADS处理稳定转染细胞,MTT观察细胞增殖的变化。用一种浓度的5-FU或DADS处理稳定转染细胞,流式细胞术观察亚G1细胞的比例。结果:免疫荧光结果表明,Bcl-XLsiRNA稳定转染组细胞中Bcl-XL蛋白的表达比阴性siRNA稳定转染组细胞Bcl-XL基因的表达均有明显的降低。当不同浓度的5-FU(13、130、1 300、13 000 mg.L-1)或DADS(20、35、50mg.L-1)处理细胞24 h后,MTT结果表明Bcl-XLsiR-NA细胞组A570吸光度值较阴性siRNA细胞组和未转染对照明显降低,细胞生长抑制率明显增高。5-FU或DADS药物的IC50值在Bcl-XLsiRNA细胞组有明显降低。使用5-FU(130 mg.L-1)或DADS(50mg.L-1)处理细胞24 h后,流式结果表明,Bcl-XL-siRNA细胞组较阴性siRNA和正常对照细胞组亚G1细胞比例有明显增高。结论:Bcl-XL siRNA下调了MGC-803细胞Bcl-XL基因的表达;Bcl-XLsiRNA能够促进MGC-803细胞凋亡,增强细胞对化疗药物的敏感性。AIM： To determine the inhibitory effect of the hairpin Bcl-XL siRNA on the expression of Bcl-XL gene in human gastric cancer cell line MGC-803, and the effect of Bcl-XL siRNA on drug sensitization in MGC-803 ceils. METHODS： Bcl-XL siRNA and negative siRNA were constructed and stably trarlsfected into MGC-803 cells. Immunofluorescence was used to detect the Bcl-XL gene expression. Drug sensitivity of the cells to 5-fluorouracil （5-FU） and diallyl disulfide （DADS） was analyzed with MTr and flow cytometry. RESULTS： Protein expression level of Bcl-XL in Bcl-XL siRNA stable transfectants was reduced to almost background level compared with negative siRNA transfectants or untreated cells. MTr results showed that the cells inhibitary rates of Bcl-XL siRNA transfectants were higher than that of negative vector or untreated cells after treated with 13, 130, 1300, and 13 000 mg· L^- 1 of 5-FU or treated wig 20,35, and 50 mg· L^-1 of DADS. IC50 value of DADS or 5-FU in Bcl-XL siRNA transfected cells was significant lower than that of negative siRNA or untreated cells. Moreover, flow cytometry results demonstrated that Bcl-XL siRNA cells showed higher sub G1 population than negative siRNA or untreated cells after addition 50 mg· L^-1 of DADS or 130 mg·L^-1 of 5-FU. CONCLUSION： siRNA targeting Bcl- XL gene can specifically down-regulate Bcl-XL expression in MGC-803 cells, and sensitize cells to 5-FU or DADS. Bcl-XL siRNA may be a potential therapy agent against human gastric adenocarcinoma.

关 键 词：小干扰RNA Bcl—XL MGC-803细胞 凋亡 DADS 5-FU 

分 类 号：R965[医药卫生—药理学]