Bcl-XL siRNA 对肺腺癌细胞A549/DDP药物敏感性及凋亡的影响

中国临床药理学与治疗学 ›› 2007, Vol. 12 ›› Issue (1): 57-61.

Bcl-XL siRNA 对肺腺癌细胞A549/DDP药物敏感性及凋亡的影响

黄泽香, 刘丽娟, 姚淑琼, 雷小勇, 朱炳阳, 唐圣松, 廖端芳

南华大学药物药理研究所, 衡阳 421001, 湖南

收稿日期:2006-11-01 修回日期:2006-12-06 出版日期:2007-01-26 发布日期:2020-10-26
通讯作者: 雷小勇, 男, 副教授, 硕士生导师, 研究方向:肿瘤药理学。Tel:0734-8282176 E-mail: leixiaoyong@yahoo.com.cn
作者简介:黄泽香, 女, 硕士生, 研究方向:肿瘤药理学。Tel:0734-8281408 　E-mail:huangzexiang0717@sina.com
基金资助:
湖南省教育厅青年基金( 03B034)

Bcl-XL siRNA sensitisizes cisplatin-resistant human lung adenocarcinoma cells A549 to cisplatin

HUANG Ze-xiang, LIU Li-juan, YAO Shu-qiong, LEI Xiao-yong, ZHU Bing-yang, TANG Sheng-song, LIAO Duan-fang

Institute of Pharmacy and Pharmacology, Nanhua University, Henyang 421001, Hunan, China

Received:2006-11-01 Revised:2006-12-06 Online:2007-01-26 Published:2020-10-26

摘要/Abstract

摘要： 目的: 探讨小干扰RNA 沉默Bcl-XL 对肺腺癌耐药细胞株A549/DDP 药物敏感性的影响及其凋亡机制。方法: 将Bcl-XL siRNA 质粒载体转染至A549/DDP 细胞, MTT 、流式细胞仪检测细胞药物敏感性的改变;丫啶橙溴化乙锭( AO/EB) 染色法检测细胞凋亡;RT-PCR 检测Bcl-XL mRNA 水平;Western-Blot 检测Bcl-XL 、caspase-3 蛋白表达的变化。结果: 分别用0.2 、2 、20 、200 μg/mL顺铂处理细胞48 h,MTT 显示转染Bcl-XL siRNA 细胞组A₅₇₀吸光度值较阴性siRNA 细胞组和未转染对照组明显降低, 细胞生长抑制率明显增高;用20 μg/mL顺铂处理细胞48 h, 流式细胞仪检测, 转染Bcl-XL siRNA 组凋亡率较转染阴性siRNA 组和未转染组增加;AO/EB 染色显示转染Bcl-XL siRNA 的细胞较转染阴性siRNA 及未转染者凋亡率增加;转染Bcl-XL siRNA 降低了Bcl-XL mRNA 和蛋白水平, 升高了caspase-3 活性形式蛋白表达。结论: Bcl-XL siRNA 特异性下调A549/DDP 细胞Bcl-XL 的表达, 活化caspase-3 蛋白, 促进A549/DDP 细胞凋亡, 增强该肺腺瘤细胞对顺铂的敏感性。

关键词: 小干扰RNA, Bcl-XL, A549/DDP 细胞, 凋亡, 顺铂

Abstract: AIM: To investigate the effect of Bcl-XL siRNA on apoptosis and drug sensitization in cisplatin-resistance human lung adenocarcinoma cell line A549/DDP and to determine the inhibitory effect of the Bcl-XL siRNA on the expression of Bcl-XL gene in cells A549/DDP. METHODS: Bcl-XL siRNA and negative siRNA plasmid vector were stably transfected into A549/DDP cells.Drug sensitivity of the cells to cisplatin ( DDP) was analyzed with MTT and flow cytometry.Spontaneous apoptosis of cells was detected by AO/EB dyeing.RT-PCR and Western-blot were used to detect the target gene expression. RESULTS: MTT results showed that Bcl-XL siRNA transfectants had a higher cell inhibition rate than negative siRNA or untreated cells after treated with 0.2, 2, 20, 200 μg/mL DDP.Moreover, flow cytometry results demonstrated that Bcl-XL siRNA cells had increased apoptosis rate after addition of 20 μg/mL DDP.Spontaneous apoptosis of cells were significantly increased in Bcl-XL siRNA stable transfectants.The mRNA and protein expression level of Bcl-XL in Bcl-XL siRNA stable transfectants were obviously reduced compared with negative siRNA transfectants or untreated cells, moreover, Bcl-XL siRNA increased the activity of active caspase-3. CONCLUSION: siRNA targeting Bcl-XL gene can specifically down-regulate Bcl-XL expression in A549 DDP cells, increase cell spontaneous apoptosis and sensitize cells to DDP.Bcl-XL siRNA may be a potential therapy agent against human lung adenocarcinoma.

Key words: siRNA, Bcl-XL, A549/DDP, apoptosis, cisplatin

中图分类号:

R966

黄泽香, 刘丽娟, 姚淑琼, 雷小勇, 朱炳阳, 唐圣松, 廖端芳. Bcl-XL siRNA 对肺腺癌细胞A549/DDP药物敏感性及凋亡的影响[J]. 中国临床药理学与治疗学, 2007, 12(1): 57-61.

HUANG Ze-xiang, LIU Li-juan, YAO Shu-qiong, LEI Xiao-yong, ZHU Bing-yang, TANG Sheng-song, LIAO Duan-fang. Bcl-XL siRNA sensitisizes cisplatin-resistant human lung adenocarcinoma cells A549 to cisplatin[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2007, 12(1): 57-61.

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