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目的 :探讨脂质体转染细胞周期素B1(cyclinB1)反义脱氧寡核苷酸 (ASON)对HL6 0细胞增殖调控的作用。方法 :用针对cyclinB1mRNA 5’端编码区起始密码子 (ATG/AUG)的ASON ,通过脂质体导入HL6 0细胞共培养后 ,用流式细胞术 (FCM)和RT -PCR分别检测cyclinB1蛋白和mRNA的表达水平 ,电镜和原位细胞凋亡检测法(POD)、FCM及DNA凝胶电泳法检测细胞凋亡。结果 :CyclinB1ASON组与SON及空白对照组相比 ,ASON能特异地抑制cyclinB1蛋白及mRNA水平的表达 ,当ASON的浓度达到一定程度时 ,HL6 0细胞的增殖及集落形成率均明显受抑制 ,出现细胞凋亡 ,并且此作用随ASON浓度的升高而增强。结论 :CyclinB1的特异ASON能封闭其蛋白及mRNA的表达水平 ,可剂量依赖性地抑制白血病细胞增殖 ,诱导细胞凋亡。
Objective: To investigate the effect of liposome-transfected cyclinB1 antisense oligodeoxynucleotides (ASON) on the proliferation of HL60 cells. Methods: The cotransfected HL60 cells were transfected with ASON encoding the initiation codon (ATG / AUG) of 5 ’end of cyclinB1 mRNA and the expression of cyclinB1 protein was detected by flow cytometry (FCM) and RT-PCR And mRNA expression were detected by electron microscopy and in situ cell apoptosis assay (POD), FCM and DNA gel electrophoresis detection of apoptosis. RESULTS: Compared with SON and blank control group, ASON could specifically inhibit the expression of cyclinB1 protein and mRNA in CyclinB1ASON group. When the concentration of ASON reached a certain level, the proliferation and colony formation rate of HL60 cells were significantly inhibited and appeared Apoptosis, and this effect with the ASON concentration increased. CONCLUSION: Specific ASON of CyclinB1 can block the expression of its protein and mRNA, and can inhibit the proliferation of leukemia cells in a dose-dependent manner and induce apoptosis.