Apg-2过表达对BaF3-MIGR1及稳定表达BCR/ABL的BaF3-P210细胞增殖的影响  

Effect of Overexpression of Apg-2 on Proliferations of BaF3-MIGR1 Cells and BaF3-P210 Cells Stably Expressing BCR / ABL

作　　者：李春莉[1] 刘钉宾[1] 袁颖[2] 彭智[1] 陶崑[1] 史梦[1] 曹唯希[1] 冯文莉[1] 

机构地区：[1]重庆医科大学医学检验性临床血液学教研室临床检验诊断学教育部重点实验室,重庆400016 [2]重庆医科大学附一院普外科,重庆400016

出　　处：《中国生物制品学杂志》2010年第2期129-133,共5页Chinese Journal of Biologicals

基　　金：教育部博士学科点专项基金(2005631007)

摘　　要：目的探讨热休克蛋白Apg-2过表达对小鼠pMIGR1空载体感染细胞BaF3-MIGR1和BCR/ABL转化细胞BaF3-P210(简称BP210)增殖的影响。方法构建pIERS2-EGFP-Apg-2真核表达质粒,电穿孔转染BaF3-MIGR1和BP210细胞,经G418筛选稳定表达细胞株,分别用RT-PCR和Western blot鉴定Apg-2的表达。采用Am-Blue法和流式细胞仪(FCM)检测Apg-2过表达对BaF3-MIGR1和BP210细胞增殖的影响,光镜观察细胞形态变化。结果pIERS2-EGFP-Apg-2真核表达质粒构建正确,转染细胞后筛选到稳定过表达Apg-2的BaF3-MIGR1-Apg-2和BP210-Apg-2细胞株,BaF3-MIGR1-Apg-2细胞的增殖速度明显低于BaF3-MIGR1细胞,BP210-Apg-2细胞比BP210细胞增殖加快。光镜下可见Apg-2过表达的两种细胞形态发生变化,瑞氏染色可见细胞核增多。结论Apg-2能够促进BCR/ABL阳性细胞的增殖,可能与慢性粒细胞白血病的发生发展相关。Objective To investigate the effect of overexpression of heat shock protein Apg-2 on the proliferations of BaF3-MIGR1 cells infected with empty vector pMIGR1 and BaF3-P210（BP210）cells transformed with BCR / ABL. Methods A eukaryotic expression vector pIERS2-EGFP-Apg-2 was constructed and transfected to BaF3-MIGR1 and BP210 cells by eletroporation. The cell strains for stable expression were screened with G418, and the expression of Apg-2 was identified by RT-PCR and Western blot. The effect of overexpression of Apg-2 on the proliferations of BaF3-MIGR1 and BP210 cells were evaluated by Am-Blue method and flow cytometry（FCM）, and the morphology of cells were observed by light microscopy. Results Recombinant plasmid pIERS2-EGFP-Apg-2 was constructed correctly. After transfection, the cell strains BaF3-MIGR1-Apg-2 and BP210-Apg-2 for stable overexpression of Apg-2 were screened. BaF3-MIGRE1-Apg-2 cells were proliferated slowly as compared with BaF3-MIGR1 cells. However, BP210-Apg-2 cells were proliferated rapidly as compared with BP210 cells. Morphological changes of the two cell strains were observed by light microscopy, and increased nucleuses by Wright staining. Conclusion Apg-2 promoted the proliferation of BCR / ABL positive cells, thus might be associated with the onset and progression of chronic myelogeneous leukemia.

关 键 词：Apg-2 慢性粒细胞白血病 BCR/ABL 细胞增殖 

分 类 号：Q789[生物学—分子生物学] Q25