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纸质出版:2008
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史梦,冯文莉,张文萍,王小中,黄世锋,曾建明,温健萍,陶昆,陈新敏,曹唯希,黄宗干.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[J].陆军军医大学学报(原第三军医大学学报),2008,30(19):1791-1794.
SHI Meng,FENG Wen-li,ZHANG Wen-ping,WANG Xiao-zhong,HUANG Shi-feng,ZENG Jian-ming,WEN Jian-ping,TAO Kun,CHEN Xin-min,CAO Wei-xi,HUANG Zong-gan.Construction of BaF3-P210 cell line stably expressing BCR/ABL protein and its biological activity[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(19):1791-1794.
史梦,冯文莉,张文萍,王小中,黄世锋,曾建明,温健萍,陶昆,陈新敏,曹唯希,黄宗干.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[J].陆军军医大学学报(原第三军医大学学报),2008,30(19):1791-1794. DOI:
SHI Meng,FENG Wen-li,ZHANG Wen-ping,WANG Xiao-zhong,HUANG Shi-feng,ZENG Jian-ming,WEN Jian-ping,TAO Kun,CHEN Xin-min,CAO Wei-xi,HUANG Zong-gan.Construction of BaF3-P210 cell line stably expressing BCR/ABL protein and its biological activity[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(19):1791-1794. DOI:
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(P0.05);G
0
/G
1
期细胞比例下降,S期和G
2
/M期细胞比例增高(P0.05);经STI571处理后的细胞增殖抑制率为(54.08±1.90)%(P0.05),早期凋亡率为(12.35±2.04)%(P0.05)。 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。
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 target cell was determined by PCR and DNA sequencing
trypan blue staining assay
flow cytometry and MTT assay. Results The bcr/abl gene was integrated into the BaF3 cell genome; RT-PCR and Western blot verified the stable expression of the bcr/abl gene and protein in the screened subclone cell line BaF3-P210. Compared with the control group
the cell proliferation rate was promoted (P0.05)
and the cell proportion in G
0
/G
1
phase was decreased with the increasing of the cell proportion in both S and G
2
/M phase (P0.05); the cell proliferation inhibition rate and the early apoptosis rate of the BaF3-P210 cells were respectively (54.08±1.90)%(P0.05) and (12.35±2.04)% (P0.05) after the treatment of protein tyrosine kinase inhibitor STI571. Conclusion The transformed mouse BaF3-P210 cell line stably expressing BCR/ABL was successfully constructed. The expression of BCR/ABL not only promoted the proliferation
but also accelerated the cell cycle progression of the BaF3 cells. It endowed the BaF3-P210 transformed cell line with the sensitivity to STI571.
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