stathmin基因的RNA干涉抑制人白血病K562细胞体外增殖的作用  

Inhibitory effect on human leukemia K562 cells by RNA interference of stathmin gene

作　　者：吴冰 白海 王存邦 王燕[2] 张惠中[2] 

机构地区：[1]兰州军区血液病研究所,甘肃兰州730050 [2]第四军医大学唐都医院中心实验室,陕西西安710038

出　　处：《现代肿瘤医学》2008年第3期329-331,共3页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(30371445)

摘　　要：目的:研究RNA干涉(RNA interference,RNAi)抑制stathmin基因表达后对人白血病K562细胞体外增殖的作用。方法:将合成的寡核苷酸链退火形成双链,连接入经BamHⅠ和HindⅢ双酶切后的pSilencer4.1-CMV neo真核表达载体。酶切及测序鉴定。脂质体法转染重组质粒入K562细胞,RT-PCR检测其对mR-NA的干涉效果;MTT比色法检测K562细胞体外增殖能力。结果:经酶切及测序鉴定,成功构建重组质粒。RT-PCR显示所构建的干涉载体成功地抑制了目的基因的转录,同时细胞增殖活性降低。结论:RNA干涉成功抑制stathmin基因表达并使人白血病K562细胞体外增殖活性明显降低。Objective：To study the RNA interference -mediated inhibition of stathmin gene on the proliferation of human leukemia K562 cells. Methods： Two target gene segments were synthesized and cloned into vector respectively to construct two recombinant eukaryotic expression vectors ： pSilencer4.1 - CMV neo - S1 and pSilencer4.1 - CMV neo - S2. The two recombinant vectors were identified by enzyme digestion analysis and DNA sequencing. Then K562 cells were transfected with pSilencer4.1 -CMV neo -S1 and pSilencer4.1 -CMV neo -S2 , then subjected to Neomycin selection. In Neomycin - resistant cells, the interference effect was detected by RT - PCR. The proliferation of transfected K562 cells was examined by MTr assay. Results： Enzyme digestion analysis and DNA sequencing showed that the target segments were cloned into pSilencer4.1 - CMV neo vector respectively. The result of RT - PCR indicated that both vectors could knock down the transcription of stathmin gene. At the same time, the growth of transfected cells was decelerated. Conclusion： stathmin gene silencing by RNA interference contributed to a distinctive inhibition of the proliferation of human leukemia K562 cells in vitro.

关 键 词：STATHMIN RNA干涉 真核表达载体 K562细胞 

分 类 号：R733.7[医药卫生—肿瘤]