Bcl-2 siRNA抑制HL-60细胞bcl-2基因表达  

Inhibition of bcl-2 gene expression by siRNA in HL-60 cells

作　　者：雷小勇[1] 燕春艳[2] 涂玉林[2] 钟苗[1] 冯兰芳[1] 廖端芳[1] 

机构地区：[1]南华大学药物药理研究所 [2]南华大学心血管疾病研究所,湖南衡阳421001

出　　处：《中国药理学通报》2005年第12期1445-1450,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30300426);湖南省教育厅青年基金资助项目(No200416)

摘　　要：目的应用RNA干扰技术观察Bcl2siRNA对白血病细胞HL60中bcl2基因表达的影响,探讨siRNA技术在白血病治疗中的作用。方法利用化学合成法体外化学合成Bcl2siRNA,将Bcl2siRNA与HL60细胞共孵育,用MTT法、荧光染色观察细胞增殖及凋亡情况,用RTPCR检测Bcl2mRNA水平,用荧光染色测Bcl2蛋白水平。结果Bcl2siRNA与HL60细胞共孵育48h后,HL60细胞凋亡率增加,Bcl2mRNA水平下调,Bcl2蛋白表达水平降低。转染GAPDHsiRNA组的细胞对Bcl2表达无影响。结论Bcl2siRNA能够特异性降低Bcl2mRNA水平及蛋白质的表达,促进HL60细胞凋亡。Aim To study the interference role of Bcl-2 siRNA on HL-60 cells. Methods Bcl-2 siRNA was synthesized in vitro transcription with silencer siRNA construction kit. The synthesized siRNA was transfected into HL-60 ceils with lipid siPORT transfection. Forty-eight hours after the transfection, we used MTT and immunofluorescence to detect cell proliferation and apoptosis, and used RT-PCR and immunofluorescence to detect the level of Bcl-2 mRNA and Bcl-2 protein expression. Result Bcl-2 siRNA reduced the level of Bcl-2 mRNA and Bcl-2 protein expression in HL-60 cells and induced cell apoptosis. There was no difference on the effect of other groups compared with the control. Effective Bcl-2 siRNA specifically degraded Bcl-2 expression in the levels of mRNA and protein and induced HL-60 ceils apoptosis. Conclusion These results indicate that siRNA is a highly specific tool for targeted gene knockdown, siRNA-mediated gene silencing is a reliable approach for large-scale screening of gene function and drug target validation.

关 键 词：RNA干扰 BCL-2 SIRNA HL-60 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R341[医药卫生—基础医学]