摘要
目的:研究hTERT基因反义寡核苷酸(ASODN)对HL-60细胞诱导凋亡作用及细胞周期的影响。方法:应用hTERT ASODN封闭HL-60细胞hTERT基因,RT-PCR方法检测目的基因的表达;流式细胞仪检测细胞凋亡及细胞周期分析,TUNEL方法检测细胞凋亡,琼脂糖凝胶电泳检测凋亡细胞DNA梯带。结果:hTERT ASODN作用细胞72h后,hTERT基因表达明显受到抑制(P<0.01)。流式细胞仪检测显示:ASODN组细胞阻滞于G0/G1期,S、G2/M期细胞减少(P<0.05);细胞增殖指数(PI)明显降低(P<0.05);检测出早期凋亡峰。Annexin V/PI检测及TUNEL检测均显示:ASODN组凋亡细胞阳性率明显高于SODN组(P<0.01)。琼脂糖凝胶电泳显示:ASODN组出现凋亡DNA梯带。结论:hTERT ASODN能够封闭目的基因表达,阻滞HL-60细胞于G0/G1期,并诱导细胞凋亡,对治疗白血病具有潜在应用价值。
Objective： To investigate the effects of hTERT antisense oligodeoxynueleotide （ASODN） on the cell cycle and the induction of apoptosis in HL-60 cells. Methods： After treatmenl with hTERT ASODN the expression of hTERT was assessed by RT-PCR and agarose gel electrophoresis, apoptosis was detected with TUNEL, and cell cycle ratios were determined using flow eylometry （FCM）. Results： After 72h of treatment with hTERT ASODN, expression of the hTERT gene was inhibited significantly （P〈0.01）. Analysis of the FCM data revealed that the ASODN trealed cells were blocked at GO/G1 phase and that Ihe number of cells in S phase and G2/M phase was reduced （P〈0.05）. The proliferation index of the ASODN-treated group was markedly decreased （P〈0.05）. In the ASODN-treated group the DNA ladder and early apoptosis peak appeared when samples were analyzed by agarose gel electrophoresis and FCM, respectively. Both Annexin V/PI FCM data and TUNEL resuhs indicaled that the number of apoptotic cells in the ASODN-treated group was much higher than Ihat found in the SODN-treated group （P〈0.01）. Conclusion： The hTERT ASODN inhibits expressima of the hTERT gene, blocks HL-60 cells at G0/GI phase, induces apoptosis and has a potential therapeutic effect for leukemia.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2007年第15期841-844,共4页
Chinese Journal of Clinical Oncology
基金
河南省医学科技创新人才工程项目基金资助(编号:2004107031)
