过表达BMI-1对HeLa细胞中HOX基因表达和细胞周期的影响  

Effects of BMI-1 over-expression on HOX family expression and cell cycle in HeLa cells

作　　者：陈凤花[1] 李一荣[1] 王琳[1] 胡丽华[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院,湖北武汉430022

出　　处：《中国病理生理杂志》2009年第12期2366-2370,共5页Chinese Journal of Pathophysiology

基　　金：湖北省科技攻关资助项目(No.2005AA304B08)

摘　　要：目的:将构建成功的真核表达载体pEGFP-BMI-1转染宫颈癌细胞系HeLa,检测其对同源盒(HOX)基因表达和细胞周期的影响。方法:采用脂质体转染法,将质粒pEGFP-BMI-1DNA瞬时转染HeLa细胞,确定融合蛋白B细胞特异性莫洛尼氏白血病毒插入位点1-加强型绿色荧光蛋白(BMI-1-EGFP)表达后,实时荧光定量RT-PCR方法检测转染前后HeLa细胞中周期素依赖性激酶抑制剂P16INK4a、人类端粒酶逆转录酶(hTERT)、同源盒A9(HOXA9)、同源盒B4(HOXB4)和同源盒C13(HOXC13)mRNA的表达变化,PI染色流式细胞仪检测细胞周期。结果:(1)在HeLa细胞中过表达BMI-1显著下调P16INK4a、HOXA9和HOXC13 mRNA的表达,分别平均降低为对照组的9.2%、10.9%和69.7%(P<0.01),而hTERT和HOXB4 mRNA的表达变化无显著差异(P>0.05)。(2)pEGFP-BMI-1转染HeLa细胞后,G1期细胞由65.68%减少至50.53%,S期细胞则由27.17%增加至39.59%(P<0.01)。结论:真核表达载体pEGFP-BMI-1转染HeLa细胞过表达外源性BMI-1,显著下调P16INK4a、HOXA9和HOXC13的表达,同时G1期细胞减少、S期细胞增加,这可能是BMI-1参与肿瘤发生发展的机制之一。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 P16INK4a, 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 P16INK4a, 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 G1 population and a significant increase from 27. 17% to 39. 59 % in S population after trans- fection （P 〈 0.01 ）. CONCLUSION： BMI- 1 over- expression in HeLa cells downregulates mRNA expressions of P16tNK4a, HOXA9 and HOXC13, decreases G1 population and increases S population. Therefore, BMI - 1 may be involved in carcinogenesis and cancer development.

关 键 词：B细胞特异性莫洛尼氏白血病毒插入位点1 周期素依赖性激酶抑制剂P16INK4a 同源盒A9 同源盒C13 

分 类 号：R363[医药卫生—病理学]