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

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.