端粒酶RNA的反义寡核苷酸诱导白血病细胞凋亡的研究

目的研究端粒酶RNA的反义寡核苷酸(As-ODN)抗白血病作用及其机制。方法采用脂质体包裹As-ODN转染入HL-60细胞；用端粒重复序列扩增法(TRAP)-酶联免疫吸附试验(ELISA）检测HL-60细胞的端粒酶活性变化；用细胞形态学观察、DNA琼脂糖凝胶电泳和流式细胞术检测凋亡细胞。结果0.1～2.0?μmol/LAs-ODN转染入HL-60细胞，培养1～6?d，HL-60细胞端粒酶活性（A值）由1.043±0.045降至0.063±0.011，且有剂量依赖性和时间依赖性。As-ODN转染的HL-60细胞培养后细胞增殖速度减慢，发生了细胞凋亡。而错义寡核苷酸(Ms-ODN)则无此效应。结论As-ODN能特异性抑制HL-60细胞的端粒酶活性,诱导细胞凋亡。

Study on induction of leukemic cell apoptosis by antisense oligodeoxynucleotide of human telomerase RNA

OBJECTIVE: To explore the antitumor activity of human telomerase RNA antisense oligodeoxynucleotide (As-ODN) to leukemic cells and its mechanisms. METHODS: As-ODN was transfected into HL-60 cells by liposomal transfection. Telomerase activity of HL-60 cells was examined by telomeric repeat amplification protocol (TRAP)-enzyme-linked immunosorbent assay (ELISA). Apoptosis was analyzed by morphology, DNA agarose gel electrophoresis and flow cytometry. RESULTS: The telomerase activity in HL-60 cells, being transfected with 0.1 - 2.0 micromol/L of As-ODN for 1 - 6 days, varied from 1.043 +/- 0.045 to 0.063 +/- 0.011 in a dose- and time-dependent manner. The proliferation of As-ODN-transfected HL-60 cells was lower than that of control, and the cells underwent apoptosis. The Ms-ODN (missense ODN)-transfected HL-60 cells didn't show these changes. CONCLUSION: As-ODN can inhibit the telomerase activity of HL-60 cells specifically and induce apoptosis.