Effects of BMI-1 over-expression on HOX family expression and cell cycle in HeLa cells
- VernacularTitle:过表达BMI-1对HeLa细胞中HOX基因表达和细胞周期的影响
- Author:
Fenghua CHEN
;
Yirong LI
;
Lin WANG
;
Lihua HU
- Publication Type:Journal Article
- Keywords:
B-cell specific moloney leukemia virus insertion site 1;
Cyclin-dependent kinase inhibitor P16~(INK4a);
Homeobox A9;
Homeobox C13
- From:Chinese Journal of Pathophysiology
2009;25(12):2366-2370
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To observe whether transfection of mammalian expression vector pEGFP containing the gene of B-cell specific moloney leukemia virus insertion site 1(BMI-1) could express in human cervix cancer cell line HeLa, and to detect its effect on HOX family expression and cell cycle.METHODS: pEGFP-BMI-1 was transfected into HeLa cells with Lipofectamine 2000. The expression of pEGFP-BMI-1 was determined by EGFP fluorescence and Western blotting. SYBR green I real-time RT-PCR was used to quantitate mRNA expression of P16~(INK4a), hTERT, HOXA9, HOXB4 and HOXC13. FACS analysis was used to detect the change of cell cycle.RESULTS: In HeLa cells transfected with pEGFP-BMI-1, the results of real-time RT-PCR showed that the mRNA expressions of P16~(INK4a), HOXA9 and HOXC13 were reduced to 9.2%, 10.9% and 69.7%, respectively, as compared to control HeLa cells (P<0.01). However, hTERT and HOXB4 mRNA expressions did not change significantly (P>0.05). FACS analysis showed a decrease from 65.68 % to 50.53% in G_1 population and a significant increase from 27.17% to 39.59 % in S population after transfection (P<0.01).CONCLUSION: BMI-1 over-expression in HeLa cells down-regulates mRNA expressions of P16~(INK4a), HOXA9 and HOXC13, decreases G_1 population and increases S population. Therefore, BMI-1 may be involved in carcinogenesis and cancer development.
