稳定表达BCR/ABL的小鼠BaF3-P210细胞株的建立及其生物学特性的研究

目的构建能稳定表达BCR／ABL的小鼠转化细胞株BaF3-P210，并初步探讨BCR／ABL对BaF3细胞生物学特性的影响。方法将含有bcr／abl基因的逆转录病毒载体转染到包装细胞，收集病毒感染BaF3细胞，经筛选和亚克隆，获得稳定表达BCR／ABL的转基因BaF3-P210细胞。用PCR、DNA测序、RT-PCR和Western blot检测bcr／abl基因在靶细胞中的整合和表达；用台盼蓝细胞计数法、流式细胞仪（FCM）和MTT法研究BCR／ABL对BaF3细胞生物学特性的影响。结果bcr／abl基因整合在BaF3细胞基因组中，BaF3-P210细胞能稳定表达bcr／abl基因及其蛋白。与对照组相比，BaF3-P210细胞增殖速度增快（P〈0．05）；G0／G1期细胞比例下降，S期和G2／M期细胞比例增高（P〈0．05）；经STI571处理后的细胞增殖抑制率为（54．08±1．90）％（P〈0．05），早期凋亡率为（12．35±2．04）％（P〈0．05）。结论成功构建的能稳定表达BCR／ABL的小鼠BaF3-P210转化细胞株具有增殖能力增强、细胞周期进程加速和对STI571敏感的恶性表型特征。

Construction of BaF3-P210 cell line stably expressing BCR/ABL protein and its biological activity

Objective To construct the transformed mouse BaF3-P210 cell line stably expressing BCR/ABL and to initially investigate the influence of BCR/ABL on the cell biological characteristics of BaF3 cell line. Methods The retroviral vector with bcr/abl gene was transfected into the packaging cell line. The BaF3 cells were infected with the collected viral supernatant. The transgenic BaF3-P210 cell line stably expressing BCR/ABL were screened and subcloned. The integration of the bcr/abl gene in the genome of the t...