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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 effect of Bmi-1 on the transcription of stem-related molecules in tumor and embryonic stem cells. Methods The murine and human Bmi-1 overexpression vectors pCI-GFP-mbmi1 and pCI-GFP-hbmi1 were constructed respectively. The transient expression of CCE and human glioma U87 cells were detected by semi-quantitative RT-PCR. The expression of CD133, Nestin and other dry molecules in Oct-4, Nanog, c-Myc, Klf4 and Sox2 as well as tumor cells were detected by clone formation assay. Results After transient transfection of Bmi-1 overexpression vector, the expression of Nanog in CCE cells was up-regulated, while the expression of Oct-4, Sox2 and Klf4 did not change significantly. The expression of Nestin and Oct-4 in U87 cells was up-regulated while the changes of CD133 and Nanog were not obvious ; Both c-Myc transcriptional expression were significantly reduced; overexpression of Bmi-1 CCE cells to promote self-renewal clone formation. Conclusion Bmi-1 is involved in transcriptional regulation of stem-related molecules in tumor and embryonic stem cells.