c-myc小干扰RNA诱导人髓系白血病细胞系HL-60细胞凋亡

本研究探讨c-myc小干扰RNA对白血病细胞系HL-60诱导凋亡的作用。将体外合成的siRNA转染HL-60细胞，观察形态学变化，应用MTT法绘制细胞生长曲线，用细胞集落培养观察c-mycsiRNA对HL-60细胞增殖的影响；应用DNA片段化分析细胞的凋亡，RT—PCR及Western blot检测c-mycsiRNA作用前后Comyc基因mRNA及蛋白表达水平的变化。结果表明：ComycsiRNA能明显抑制HL-60细胞增殖，半数抑制浓度（IC50）约为150nmol／L；DNA片段化的检出证实C-mycsiRNA能有效诱导HL-60细胞调亡，凋亡率与药物作用时间呈正相关。C-mycsiRNA与HL-60细胞作用后C-myc基因和蛋白的表达水平与作用时间呈负相关。结论：C-mycsiRNA能有效抑制HL-60细胞增殖，降低C-myc基因和蛋白的表达，诱导其凋亡。

Apoptosis of Human Myeloid Leukemia Cell Line HL-60 Cells Induced by siRNA Targeting Gene c-myc

This study was purposed to investigate the effects of siRNA targeting c-myc on apoptosis induction, proliferationin inhibition as well as c-myc protein and mRNA expression in human myeloid leukemia ceU line HL-60 cells. C-myc siRNA synthesized in vitro was transfected into HL-60 cells by liposome. Changes of cell morphology were observed . Growth inhibition was detected by MTT assay and colony formation assay, and cell apoptosis was determined by DNA ladder. The expressions of c-myc mRNA and protein were detected by RT-PCR and Western-blot respectively. The results indicaded that c-myc siRNA remarkably inhibited the cell proliferation, with an IC50 value of 150 nmol/L. Data of DNA ladder showed that HL-60 cells apoptosis could be efficiently induced by c-myc siRNA, the apoptosis rate positively correlated with the time duration of treatment with drugs. The c-myc mRNA and protein expressions on HL-60 cells decreased after treatment with c-myc siRNA, which negatively correlated with time duration of treatment. It is concluded that c-myc siRNA can efficiently induce growth inhibition, decrease the expressions of c-myc mRNA and protein, and induce apoptosis in HL-60 cells.