脂质体包裹DNMT1反义寡核苷酸诱导白血病细胞株孕酮受体去甲基化的实验研究

Demethylation of progesterone receptor in leukemia cell line induced by DNMT1 antisense oligonucleotide wrapped in liposome

目的 研究脂质体包裹DNA甲基转移酶 (DNMT1)反义寡核苷酸转染对白血病细胞株孕酮受体 (ProgesteroneReceptor,PR)启动子甲基化及表达的影响 ,探讨甲基化基因治疗的新途径。方法 人工合成的DNMT1反义寡核苷酸MG88用脂质体包裹后转染白血病细胞株 ,用RT PCR检测DNMT1、PRBmRNA表达的变化以及用MSP分析PRA、PRB甲基化状态的改变。结果 用MG88转染白血病细胞株后 ,DNMT1mRNA显著下降 ;PRA、PRB启动子出现脱甲基化 ,PRBmRNA表达显著上升。结论 DNMT1反义寡核苷酸MG88可以诱导白血病细胞PR脱甲基化 ,使PRBmRNA表达升高 ,为DNA甲基化基因治疗的实验性研究提供了可能。

Objective To investigate the effects of the transfection of DNMT1 antisense oligonucleotide wrapped in liposome on the promotor methylation and expression of progesterone receptor in leukemia cell lines so as to find a new pathway of methylation for gene therapy. Methods Artificially synthesized DNMT1 antisense oligonucleotide MG88 wrapped in liposome was transfected into the leukemia cell line. Changes of DNMT1 & PRB mRNA were detected by RT PCR, and the changes of the methylation status of PRA & PRB were analyzed by MSP. Results After transfection of leukemia cell line with MG88, DNMT1 mRNA decreased significantly. PRA & PRB promotors were demethylated, and the expressions of PRB mRNA increased significantly. Conclusion MG88 can induce the demethylation of PR promoters in leukemia cell line, resulting in the increase of PRB mRNA expression. So MG88 can be used in the experimental study of DNA methylation for gene therapy.