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

摘要

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 realtime 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.%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.