采用优化TRAP方法检测乳腺癌细胞系中端粒酶活性  

Detection of telomerase activity in breast cancer cell lines using optimized TRAP method

作　　者：王皓莉 周建文[2] 彭娅娅 蔡美虹 王维 WANG Hao-li;ZHOU Jian-wen;PENG Ya-ya;CAI Mei-hong;WANG Wei(Department of Reproductive Health and Infertility,Guangzhou First People’s Hospital,School of Medicine,South China University of Technology,Guangzhou,Guangdong 510180,China;不详)

机构地区：[1]广州市第一人民医院生殖健康与不孕症专科,华南理工大学医学院,广东广州510180 [2]中山大学附属第一医院分子诊断与基因检测中心,广东广州510080

出　　处：《中国卫生检验杂志》2021年第11期1296-1299,共4页Chinese Journal of Health Laboratory Technology

基　　金：广州市卫生健康科技西医类一般引导项目(20191A011009)。

摘　　要：目的端粒酶活性的检测方法-TRAP技术被广泛应用于肿瘤相关研究。方法用293T细胞优化端粒酶活性反应体系,并用优化的TRAP方法检测3种乳腺癌细胞系(MCF-7、MDA-MB-231和SK-BR-3)的端粒酶活性。结果293T细胞端粒酶活性随蛋白含量的降低逐渐下降。当反应体系中加入含10 ng蛋白的细胞裂解液时,尚有明显的PCR扩增条带。3种乳腺癌细胞系端粒酶活性随着反应体系中蛋白浓度的下降而下降。MDA-MB-231端粒酶活性最高,MCF-7次之,SK-BR-3端粒酶活性最低。结论优化了TRAP端粒酶活性检测方法并检测3种乳腺癌细胞系的端粒酶活性。优化的TRAP方法具有更高的敏感性并且降低了检测成本,可被广泛应用于肿瘤细胞及永生化细胞端粒酶活性的检测。Objective TRAP technique,a method for detecting telomerase activity,has been widely used in tumor-related research.Methods We optimized telomerase lysis and reaction buffer using 293 T cells.Then,we used optimized TRAP method to detect telomerase activity in three breast cancer cell lines(MCF-7,MDA-MB-231 and SK-BR-3).Results Telomerase activity of 293 T cells gradually decreased when protein content dropped.When 10 ng of cell lysate was added to the reaction system,significant PCR amplified bands were still observed.Telomerase activity in three breast cancer cell lines decreased with protein content decreasing in cell lysate.MDA-MB-231 cells had the highest telomerase activity,followed by MCF-7,and lowest telomerase activity in SK-BR-3.Conclusion We improved TRAP method and used it to detect the telomerase activity in three breast cancer cell lines.The optimized TRAP method improves sensitivity and reduces detection cost.It can be widely used to detect telomerase activity in tumor cells and immortalized cells.

关 键 词：端粒酶活性 TRAP 乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤]