Bmi-1调节干性相关分子转录的初步分析  

Transcriptional regulatory effects of Bmi-1 on stemness-associated genes in embryonic stem cells

作　　者：阮艳[1] 蹇锐[1] 程小星[2] 牛翰婕[2] 周恒宇[3] 郑宏庭[3] 胡福泉[1] 

机构地区：[1]第三军医大学基础医学部微生物学教研室,重庆市微生物工程实验室,重庆400038 [2]解放军总医院309临床部,北京100091 [3]第三军医大学新桥医院内分泌科,重庆400037

出　　处：《第三军医大学学报》2009年第9期768-771,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30700418,30700384)~~

摘　　要：目的探讨Bmi-1对肿瘤及胚胎干细胞中干性相关分子转录表达的影响。方法分别构建鼠源和人源Bmi-1过表达载体pCI-GFP-mbmi1和pCI-GFP-hbmi1;瞬时转染鼠ES细胞CCE和人神经胶质瘤细胞U87;半定量RT-PCR检测胚胎干细胞中Oct-4、Nanog、c-Myc、Klf4和Sox2及肿瘤细胞中CD133、Nestin等干性分子的表达;克隆形成实验检测CCE细胞自我更新克隆形成能力。结果瞬时转染Bmi-1过表达载体后,CCE细胞中Nanog的表达上调,Oct-4、Sox2和Klf4的表达无明显变化;U87细胞中Nestin和Oct-4表达上调,CD133、Nanog变化不明显;两者c-Myc的转录表达均显著降低;过表达Bmi-1促进CCE细胞自我更新克隆形成。结论Bmi-1参与了对肿瘤及胚胎干细胞中干性相关分子的转录调节。Objective To investigate the transcriptional polycomb group, on stemness-associated genes in cancer and regulatory effects of Bmi-1, a member of embryonic stem cells （CSCs and ESCs ）. Methods Eukaryotic expression vectors pCI-GFP-mbmi and pCI-GFP-hbmi were constructed and then transiently transfected into mouse ESCs, CCE cells and human glioma cell line, U87 cells respectively. Semi- quantitative RT-PCR was used to detect mRNA expressions of sternness factors, such as Oct-4, Nanog, c-Myc, Klf4 and Sox2 in CCE cells, and expressions of Bmi-1, Oct-4, Nanog, c-Myc, CD133 and Nestin in U87 cells. Clone formation assay was used to elucidate the potential to form self-renewal clones of CCE cells. Results Overexpression of Bmi-1 in CCE cells up-regulated the expression of Nanog. The expressions of other sternness factors such as Oct-4, Sox2 and Klf4 had no significant change. U87 ceils transiently transfected with Bmi-1 promoted the expression of Oct-4 and Nestin. The expression level of c-Myc was reduced in the 2 kinds of ceils. Overexpression of Bmi-1 promoted the formation of self-renewal clones in CCE cells. Conclusion Bmi-1 is involved in the transcriptional regulation of stemness-associated genes in CSCs and ESCs.

关 键 词：BMI-1 干细胞 干性相关分子 转录调控 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R394-33[医药卫生—基础医学]