Bcl-XL siRNA 对人胃腺癌 MGC-803 细胞药物敏感性的影响

中国临床药理学与治疗学 ›› 2006, Vol. 11 ›› Issue (1): 33-38.

Bcl-XL siRNA 对人胃腺癌 MGC-803 细胞药物敏感性的影响

钟苗, 雷小勇, 冯兰芳, 朱炳阳, 唐圣松, 廖端芳

南华大学生命科学与技术学院药物药理研究所, 衡阳 421001, 湖南

收稿日期:2005-10-14 修回日期:2005-11-26 出版日期:2006-01-06 发布日期:2020-10-23
通讯作者: 雷小勇, 通讯作者, 男, 博士, 硕士生导师, 副教授, 研究方向为肿瘤药理学。 Tel:0734-8282624 E-mail:lei xiaoyong@yahoo .com .cn
基金资助:
国家自然科学基金项目( №30300426 ) ;湖南省教育厅青年基金 ( №03B034)

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

ZHONG Miao, LEI Xiao-yong, FENG Lan-fang, ZHU Bing-yang, TANG Sheng-song, LIAO Duan-fang

Institute of Pharmacy and Pharmacology , College of Life Science and Technology , Nanhua University, Hengyang 421001 , Hunan, China

Received:2005-10-14 Revised:2005-11-26 Online:2006-01-06 Published:2020-10-23

摘要/Abstract

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 、50 mg·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 ( 50 mg·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。

关键词: 小干扰 RNA ;Bcl-XL;MGC-803 细胞;凋亡 , DADS ;5-FU

Abstract: 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 cells.METHODS:Bcl-XL siRNA and negative siRNA were constructed and stably transfected into MGC-803 cells .Immunofluorescence was used to detect the Bcl-XL gene expression .Drug sensitivity of the cells to 5-fluorou- racil ( 5-FU) and diallyl disulfide ( DADS) was analyzed with MTT and flow cytometry .RESULTS:Protein ex- pression level of Bcl-XL in Bcl-XL siRNA stable transfec- tants was reduced to almost background level compared with negative siRNA transfectants or untreated cells .MTT results showed that the cells inhibitary rates of Bcl-XL siRNA transfectants were higher than that of negative vec- tor or untreated cells after treated with 13, 130, 1300, and 13 000 mg·L-1 of 5-FU or treated with 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.

Key words: Small interference RNA , Bcl-XL , MGC- 803 cells , apoptosis , 5-FU , DADS

中图分类号:

R965

钟苗, 雷小勇, 冯兰芳, 朱炳阳, 唐圣松, 廖端芳. Bcl-XL siRNA 对人胃腺癌 MGC-803 细胞药物敏感性的影响[J]. 中国临床药理学与治疗学, 2006, 11(1): 33-38.

ZHONG Miao, LEI Xiao-yong, FENG Lan-fang, ZHU Bing-yang, TANG Sheng-song, LIAO Duan-fang. Bcl-XL siRNA sensitizes human MGC-803 gastric cancer cells to 5-flu- orouracil and diallyl disulfide[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2006, 11(1): 33-38.

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