外周血淋巴细胞中端粒酶活性激活及机制的研究

Research of Telomerase Activity in Lymphocytes of Human Peripheral Blood

实验采用PHA和rhIL-2激活人外周血淋巴细胞,再以流式细胞术(FCM)分析细胞周期和细胞分裂增殖;以TRAP检测外周血淋巴细胞活化前后端粒酶活性的变化;以Western blot检测hTERT和相关蛋白的表达;以荧光定量PCR分析hTERT的mRNA变化水平.结果表明:外周血淋巴细胞在被PHA和rhIL-2刺激活化后,端粒酶活性升高,升高的端粒酶活性能够被JAK抑制剂所抑制,提示端粒酶活性的升高依赖JAK信号通路.升高的端粒酶活性是由于hTERT蛋白表达的增高,而hTERT表达增高的原因是由于hTERT的mRNA表达水平升高,实验还表明这些活动都同样依赖于JAK信号通路.本研究深化了对端粒酶活性以及hTERT调控机制的认识.

PHA and rhIL-2 were adopted to stimulate human peripheral blood lymphocytes. The cell cycle and cell proliferation were analyzed based on the flowing cytometry （FCM）. By using TRAP, the changes of telomerase activity in peripheral blood lymphocytes before and after activation were detected. By using Western blot, the expression of hTERT and related protein were detected. By using the fluorescent quantitative PCR, the mRNA levels of hTERT were analyzed. The results showed that after the peripheral blood lymphocytes were activated by PHA and rhlL-2, telomerase activity was elevated, and elevated telomerase activity could be restrained by the JAK inhibitor. These results indicated that the increase of the telomerase activity depends on JAK signal pathway. Elevated telomerase activity is due to the increase of hTERT protein expression, while the hTERT is due to the increase of mRNA expression levels of hTERT. The results of the experiments showed that these activities also depends on the JAK signal pathway. Evidences about the telomerase activity and hTERT regulation mechanism are provided based on this study.