探讨端粒酶在沙利度胺诱导U266细胞增殖抑制及凋亡中的意义

Role of telomerase on proliferation and apoptosis of U266 cells induced by thalidomide

摘要:

目的：探讨端粒酶活性及人端粒酶逆转录酶（human telomerase reverse transcriptase，hTERT）mRNA的变化在监测沙利度胺诱导多发性骨髓瘤（multiple myeloma，MM）U266细胞增殖抑制、凋亡中的意义。方法：CCK-8法测定沙利度胺对U266细胞增殖抑制作用；流式细胞仪检测U266细胞凋亡率；端粒重复序列扩增酶联免疫吸附实验（TRAP-ELISA）检测端粒酶活性；逆转录聚合酶链反应检测hTERT mRNA表达水平。结果：沙利度胺对U266细胞有增殖抑制和促凋亡作用，且均呈时间-浓度依赖性；10 μg/ml沙利度胺作用24 h和48 h，端粒酶活性值分别为（7.96±0.09）和（7.65±0.07），相比对照组均无明显变化（P=0.082 0），hTERT mRNA表达分别为（2.82±0.03）和（2.31±0.04），与对照组相比表达均下降，差异有统计学意义（P=0.033 0）；50 μg/ml和100 μg/ml沙利度胺作用U266细胞24、48 h及72 h，端粒酶活性、hTERT mRNA表达分别与对照组比较表达均下降，差异有统计学意义（P=0.002 8）。结论：沙利度胺诱导U266细胞增殖抑制、凋亡与端粒酶有关；与端粒酶活性相比，hTERT mRNA改变更灵敏，可作为沙利度胺治疗MM疗效判定指标之一。

Abstract:

Objective：To investigate significances of variation of telomerase activity and human telomerase reverse transcriptase （hTERT） mRNA in monitoring the inhibition of multiple myeloma（MM） U266 cell proliferation and apoptosis as a result of thalido－mide. Methods：Inhibition of thalidomide on cell proliferation was determined by CCK-8 assay. Percentage of U266 cell apoptosis was measured by flow cytometry. Telomerase activity of U266 cells was performed by TRAP-ELISA. RT-PCR was used to detect the ex－pressions of hTERT mRNA. Results：Thalidomide inhibited the growth of U266 cells and induced cells apoptosis in a time-dose dependent manner. Cells were treated with 10 μg/ml thalidomide for 24 h and 48 h；telomerase activities of U266 cells were respec－tively 7.96±0.09 and 7.65±0.07，showing no significant difference compared with those in control group（P =0.082 0）. Expressions of hTERT mRNA were 2.82±0.03 and 2.31±0.04 respectively，significantly different from those in control group（P=0.033 0）. Telom－erase activities and hTERT mRNA expressions of 50 μg/ml and 100 μg/ml groups were significantly different from those in con－trol group（P=0.002 8）. Conclusions：Thalidomide can inhibit U266 cell proliferation and induce its apoptosis，the mechanism of which is related with telomerase. Change is more sensitive in hTERT mRNA expression than in telomerase activity and it is an ef－fective index when treating MM with thalidomid.