实时定量RT-PCR检测K562/A02细胞株bcr-abl融合基因mRNA水平  

Detection of bcr-abl Fusion Gene mRNA Level in K562/A02 Cell Line by Real-time Quantitative RT-PCR

作　　者：陈宝安[1] 周桂娜[1] 程坚[1] 乔纯[2] 吴雨洁[2] 李建勇[2] 丁家华[1] 高冲[1] 赵刚[1] 王骏[1] 鲍文[1] 宋慧慧[1] 

机构地区：[1]东南大学医学院附属中大医院血液科,江苏南京210009 [2]江苏省人民医院血液科,江苏南京210029

出　　处：《中国实验血液学杂志》2011年第1期40-43,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金资助;编号39970832;30740062

摘　　要：本研究定量分析慢性髓系白血病耐阿霉素细胞株K562/A02细胞中bcr-abl融合基因的mRNA水平。收集对数生长期的K562/A02细胞,用TRIzoL提取RNA,用实时定量RT-PCR(RQ-RT-PCR)技术检测bcr-abl融合基因及内参基因abl mRNA表达水平。结果表明:RQ-RT-PCR技术分析103-107拷贝数的重复性良好,灵敏度达10-5;K562/A02细胞中bcr-abl融合基因为高表达,bcr-abl mRNA表达量大于100%。结论:实时定量PCR检测K562/A02细胞bcr/abl融合基因mRNA水平,敏感可靠,重复性好,有助于临床辅助监测慢性髓系白血病。This study was aimed to quantitively analyze the mRNA level of bcr-abl fusion gene in K562/A02 cell line by real-time quantitative reverse transcriptase polymerase chain reaction（RQ-RT-PCR）technique.After being cultured for a period of time,the K562/A02 cell line was collected and RNA was extracted using TRIzoL kit.The real-time quantitative reverse transcriptase polymerase chain reaction technology was used to detect the level of bcr-abl fusion gene and internal reference abl gene.The results showed that a fine reproducibility was obtained between 107 and 103 copies/ml,reproducible sensitivity of RQ-RT-PCR was 10-5.The expression of bcr-abl fusion gene in K562/A02 cells was higher and the level of bcr-abl mRNA was more than 100% in K562/A02 cells.It is concluded that RQ-RT-PCR is a reliable,sensitive and reproducible method for detecting mRNA level of bcr-abl fusion gene,which may be useful in monitoring the chronic myeloid leukemia.

关 键 词：K562/A02细胞 BCR-ABL融合基因 RQ-RT-PCR 

分 类 号：R733.72[医药卫生—肿瘤]