Sun Ling, Wang Feng, Sun Hui, Le Xiao-Ping, Liu Lin-Xiang, Liu Yan-Fang, Wang Fang, Wang Yi-Hao, Ma Hong-Yan, Zhang Qin-Xian
Department of Hematology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2005 Aug;13(4):605-9.
To investigate the effects of c-myc antisense oligodeoxynucleotide (ASODN) on the telomerase activity and the induction of apoptosis in HL-60 cells, and to explore the relationship between the telomerase activity and the expression of c-myc gene in HL-60 cells, after treatment by c-myc ASODN, the expression of c-myc was detected by RT-PCR, the apoptosis, cell cycle were detected with agarose gel electrophoresis and flow cytomety, and the telomerase activity was determined with TRAP-ELISA. The results showed that after blocking c-myc gene with ASODN for 72 hours, it is obvious that the expression of c-myc gene was inhibited. The percentage of S phase HL-60 cells decreased from 55.6% to 30%, the early apoptosis peak appeared (the percentage of apoptosis cells were 25.2%) and the DNA ladders were shown. OD(450 - 690) were 2.648 +/- 0.42, 2.324 +/- 0.36, 2.162 +/- 0.38, 1.952 +/- 0.14, 1.805 +/- 0.40, 1.616 +/- 0.41 and 2.466 +/- 0.29, respectively, as the cells were treated with 0, 1, 2, 3, 4, 5 micromol/L ASODN and 5 micromol/L SODN for 72 hours. The difference was significant when compared 3, 4, 5 micromol/L groups with 0 micromol/L ASODN group respectively (P < 0.05), but the difference was no significant when compared 1, 2 micromol/L ASODN and 5 micromol/L SODN groups with 0 micromol/L ASODN group (P > 0.05). It is concluded that the c-myc gene ASODN may induce the apoptosis of cells, inhibit cells from G(1) phase into S phase and regulate the telomerase activity down in HL-60 cells by blocking the expression of c-myc gene.
为研究c-myc反义寡脱氧核苷酸(ASODN)对HL-60细胞端粒酶活性及诱导凋亡的影响,探讨HL-60细胞中端粒酶活性与c-myc基因表达的关系,用c-myc ASODN处理后,采用RT-PCR检测c-myc的表达,用琼脂糖凝胶电泳和流式细胞术检测凋亡及细胞周期,用TRAP-ELISA法测定端粒酶活性。结果显示,用ASODN封闭c-myc基因72小时后,c-myc基因的表达明显受到抑制。HL-60细胞S期的百分比从55.6%降至30%,出现早期凋亡峰(凋亡细胞百分比为25.2%)并出现DNA梯形条带。当细胞分别用0、1、2、3、4、5 μmol/L ASODN和5 μmol/L SODN处理72小时后,OD(450 - 690)分别为2.648±0.42、2.324±0.36、2.162±0.38、1.952±0.14、1.805±0.40、1.616±0.41和2.466±0.29。3、4、5 μmol/L组分别与0 μmol/L ASODN组比较差异有统计学意义(P < 0.05),但1、2 μmol/L ASODN组和5 μmol/L SODN组与0 μmol/L ASODN组比较差异无统计学意义(P > 0.05)。结论:c-myc基因ASODN可能通过封闭c-myc基因的表达诱导HL-60细胞凋亡,抑制细胞从G(1)期进入S期并下调端粒酶活性。