高表达人核糖核酸酶抑制因子的乳腺癌细胞株的建立及其鉴定  

Establishment and identification of breast cancer cell line with over-expressed human ribonuclease inhibitor

作　　者：樊建慧[1] 刘鹏[1] 崔秀云[1] 田余祥[1] 

机构地区：[1]大连医科大学生物化学与分子生物学教研室,辽宁大连116044

出　　处：《中国微生态学杂志》2008年第3期203-204,共2页Chinese Journal of Microecology

基　　金：国家自然基金资助项目(项目编号:39870242)

摘　　要：目的建立稳定高表达人核糖核酸酶抑制因子(hRI)的真核细胞系:乳腺癌细胞系,为进一步研究hRI抗肿瘤、抗氧化的作用机制奠定实验基础。方法将hRI通过逆转录病毒载体(pLNCX-hRI)经过病毒包装细胞(PA317)包装后的高滴度病毒上清,感染乳腺癌(MCF-7)细胞系,经G418筛选后,运用RT-PCR、Western blot等方法进行鉴定hRI在MCF-7中的高表达。结果hRI克隆到乳腺癌细胞(MCF-7)基因组,并随着基因组稳定高表达hRI。结论利用逆转录病毒载体感染真核细胞后获得高表达hRI的肿瘤细胞株,从而为进一步研究真核细胞内hRI的抗肿瘤作用机制提供条件。Objective To establish a stable breast cancer cell line with over expressed human ribonuclease inhibtor （hRI）, and give a base on studying the anti-tumor and anti-oxidative function of hRI. Methods An infection of hRI gene into human breast carcinoma cell line： MCF-7, by the retroviral packaging cell line PA317 which carrying the pLNCX-hRI ,then MCF-7/pLNCX-hRI cell line with a stably higher expression of hRI was selected by G418 ,and then detected the over expression of hRI by RT-PCR, Western blot, et al. Results The study showed that hRI gene integrated into the ge- nome of the MCF-7 cell,and also hRI over-expressed in the infected MCF-7 cell line with its own genome. Conclusion The results suggest that hRI integrated into the MCF-7 cell line successfully, and the establishment of MCF-7/hRI cell line provided the condition for the further study of the mechanism of anti-tumor function of hRI.

关 键 词：人核糖核酸酶抑制因子 逆转录病毒 乳腺癌细胞 

分 类 号：Q26[生物学—细胞生物学]