论文部分内容阅读
目的 探讨DNA甲基转移酶抑制剂 5 氮杂 2′ 脱氧胞嘧啶 (CdR)与组蛋白去乙酰化酶(HDAC)抑制剂丁酸钠 (SB)联合诱导白血病细胞p1 5INK4B基因重新表达的可能性。方法 采用急性髓系白血病细胞系KG1a和 2例原代培养白血病细胞为研究对象 ;限制性内切酶联合PCR技术检测p1 5INK4B基因启动子区甲基化状态 ;逆转录 聚合酶链反应 (RT PCR)检测p1 5INK4BmRNA的表达 ;Western免疫印迹法检测p1 5INK4B蛋白的表达。结果 受检细胞p1 5INK4B基因启动子区呈高甲基化状态 ,mRNA及蛋白表达完全或部分缺失 ;CdR和SB可单独诱导p1 5INK4B mRNA及蛋白表达 ,且有剂量依赖性 ;低浓度CdR(0 5 μmol/L)联合SB(0 5mmol/L)显著诱导p1 5INK4B 表达 ,其作用高于单用高浓度CdR(1 μmol/L) 或高浓度SB(1mmol/L)。结论 CdR和SB可诱导甲基化失活的白血病细胞p1 5INK4B基因重新表达 ,二者有明显的协同作用
Objective To investigate the possibility of DNA methyltransferase inhibitor 5-azacytidine 2’-deoxycytidine (CdR) combined with histone deacetylase (HDAC) inhibitor sodium butyrate (SB) in the re-expression of p15INK4B in leukemia cells . Methods Acute myeloid leukemia cell line KG1a and 2 primary cultured leukemia cells were used as research objects. Methylation status of p1 5INK4B gene promoter region was detected by restriction endonuclease assay and PCR. Reverse transcription polymerase chain reaction (RT PCR) ) To detect p1 5INK4BmRNA expression; Western immunoblotting assay p1 5INK4B protein expression. Results The promoter region of p15INK4B gene was hypermethylated and the mRNA and protein were completely or partially deleted. CdR and SB could induce the expression of p1 5INK4B mRNA and protein in a dose-dependent manner. Low concentrations of CdR (0 5 μmol / L) combined with SB (0 5 mmol / L) significantly induced the expression of p15INK4B, which was higher than that of single concentration of CdR (1 μmol / L) or high concentration of SB (1 mmol / L). Conclusion Both CdR and SB can induce the re-expression of p15INK4B gene in methylated inactivated leukemia cells, which have obvious synergistic effects