端粒酶活性的抑制对顺铂诱导HL-60细胞凋亡的影响

Effect of Inhibition of Telomerase on Cisplatin-Induced Apoptosis in HL-60 Cells

采用端粒酶聚合酶链反应 酶联免疫吸附测定 (PCRELISA)的方法 ,检测人类端粒酶逆转录酶 (hTERT)基因反义寡脱氧核苷酸 (ASODN)处理HL 60细胞前后端粒酶活性的改变 ,并进一步通过姬姆萨染色及流式细胞术方法 ,分析HL 60细胞的端粒酶活性受到抑制后顺铂对HL 60细胞凋亡的影响。结果显示 :hTERT基因ASODN作用于HL 60细胞后 ,端粒酶活性表现逐渐下降 ;hTERTASODN与顺铂联合可诱导HL 60细胞出现一系列特征性的凋亡细胞的形态学变化 ;hTERT基因ASODN作用于HL 60细胞 2 4小时再加入顺铂 ,用流式细胞术测定凋亡细胞的百分率明显高于正义寡核苷酸 (SODN)联合顺铂作用组和单用顺铂作用组 (P <0 .0 1)。然而 ,hTERT基因的ASODN下调HL 60细胞的端粒酶活性以后 ,再用DNR和Ara c处理不能加速HL 60细胞的凋亡。以上的结果表明了端粒酶活性的下调可选择性促进顺铂诱导HL

To explore the effect of inhibition of telomerase activity on cisplatin-induced apoptosis in HL-60 cells, polymerase chain reaction-enzyme-linked immunosorbent assay(PCR-ELISA) was used to determine telomerase activity in HL-60 cells untreated or treated with human telomerase reverse transcriptase(hTERT) gene antisense oligodeoxynucleotide(ASODN); apoptosis was detected by morphological observation and cell cycle analysis by flow cytomertry. Telomerase activity in HL-60 cells treated with hTERT ASODN was shown to decrease with time of treatment. The combination of hTERT ASODN and cisplatin could induce apoptosis of HL-60 cells. Morphological characteristics of apoptosis, such as cell shrinkage, chromatin condensation, nuclear segmentation and formation of apoptotic bodies, were observed. The percentage of apoptotic HL-60 cells treated with cisplatin for 48 or 72 hours after 24 hours of exposure to ASODN was significantly increased than that of groups with sense oligodeoxynucleotide(SODN) plus cisplatin or cisplatin alone(P<0.01), respectively. However, there was no difference in percentage of apoptotic HL-60 cells between hTERT ASODN plus DNR or Ara-c, SODN plus DNR or Ara-c, and DNR or Ara-c alone, respectively. The results here demonstrated that inhibition of telomerase activity by hTERT ASODN could enhance the cisplatin-induced apoptosis of HL-60 cells.