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为了探讨脂质体转染细胞周期蛋白 (cyclin)G1反义脱氧寡核苷酸 (ASON)对HL 6 0细胞增殖调控的作用 ,用针对cyclinG1mRNA 5′端编码区起始密码子 (ATG/AUG)的ASON ,通过脂质体导入HL 6 0细胞共培养后 ,用免疫组织化学法和RT PCR分别检测cyclinG1和mRNA水平的表达 ,用电镜、细胞原位凋亡检测法 (POD)、流式细胞术(FCM)及DNA凝胶电泳等方法检测细胞凋亡。结果表明 :cyclinG1ASON组与SON及空白对照组相比 ,ASON能特异地抑制cyclinG1及mRNA水平的表达 ,当ASON的浓度达到一定程度时 ,HL 6 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 :cyclinG1的特异反义脱氧寡核苷酸能封闭其蛋白及mRNA水平的表达 ,对白血病细胞的增殖有抑制作用 ,并可促使细胞凋亡 ,且有浓度依赖性。
In order to investigate the effect of liposome-transfected cyclin G1 antisense oligodeoxynucleotide (ASON) on the proliferation of HL-60 cells, the expression of start codon (ATG / AUG) ) Were transfected into HL-60 cells by lipofectamine. The expressions of cyclin G1 and mRNA were detected by immunohistochemistry and RT-PCR respectively. The expression of cyclin G1 and mRNA was detected by electron microscopy, cell in situ apoptosis assay (POD), flow cytometry Cell apoptosis (FCM) and DNA gel electrophoresis were used to detect apoptosis. The results showed that ASON could specifically inhibit the expression of cyclin G1 and mRNA in cyclin G1ASON group compared with SON and blank control group. When ASON concentration reached a certain level, the proliferation and colony formation rate of HL 60 cells were significantly inhibited, Apoptosis occurred, and this effect increased with increasing ASON concentration. Conclusion: The specific antisense oligodeoxynucleotides of cyclinG1 can block the expression of its protein and mRNA level, inhibit the proliferation of leukemia cells and induce apoptosis in a concentration - dependent manner.