摘要
目的 探讨端粒酶RNA(hTR)反义寡核苷酸 (ASODN)对急性白血病 (AL)原代细胞端粒酶活性及细胞凋亡的影响。方法 应用与人hTR起始密码子互补的硫代ASODN处理AL原代细胞 ,用台盼蓝拒染的方法计数活细胞 ,用端粒酶重复扩增分析 (TRAP) -PCR -ELISA检测法观察端粒酶活性的变化 ,用Giemsa染色、Hoechst332 5 8+PI双染荧光显微镜检查及流式细胞仪检测凋亡细胞的发生。结果 经hTR -ASODN作用后 ,AL原代细胞端粒酶活性明显受到抑制 ,并与ASODN的浓度和处理时间相关 ,10 μmol/LASODN在作用 72h时端粒酶活性为( 0 0 95± 0 0 6 ) ,较细胞对照组 ( 1 15 4± 0 15 )下调明显 (P <0 0 1) ;在作用第 5天经荧光显微镜可观察到ASODN组细胞凋亡形态学变化。流式细胞仪检测到其凋亡率为 ( 31 72± 8 6 4 ) ,与细胞对照组的凋亡率 ( 6 33± 0 91)比较 ,差异有显著性 (P <0 0 1)。结论 hTR -ASODN可明显抑制AL原代细胞端粒酶活性并诱导其凋亡 。
Objective To evalutae the effect of hTR ASODN on telomerase activity and apoptosis in primary acute leukemia(AL) cells. Methods Primary AL cells were treated with phosphorothioate ASODN complementary to the template region of hTR in vivo. The change in telomerase activity was detemined by polymerase chain reaction enzyme-linked immunoassay(PCR-ELISA). Apotosis was analyzed by Hoechst 33258+PI and flow cytometry. Results Telomerase activity was significantly suppresssed by ASODN and the effect was correlated with concentrations of ASODN and treating time. Apoptosis rates of primary AL cells treated with ASODN for 5 day significantly increased. But similar effect was not observed in SODN group. Conclusion ASODN complementary to the region of hTR can significantly suppress the telomerase activity and induce cell apoptosis. Telomerase may play an important role in the carcinogensis of leukemia and can be a new target of treatment for leukemia.
出处
《广东医学》
CAS
CSCD
2004年第12期1379-1381,共3页
Guangdong Medical Journal
基金
广东省自然科学基金资助项目 (编号 :0 43 0 0 90 2 )
广州市重点科技项目 (编号 :2 0 0 1-Z -0 3 7-0 1)
