【摘要】 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更多还原

【Abstract】 Objective To explore the possibility of re expression of silenced p15 INK4B gene in leukemia cells by 5 Aza 2′ deoxycytidine(CdR), a DNA methyltransferase inhibitor, and sodium butyrate(SB), a histone deacetylase inhibitor Methods A myeloid leukemia cell line, KG1a and two primary cultured leukemia cells isolated from bone marrow of AML M 2 and MDS RAEB T patients were chosen as the experimental target The methylation pattern of p15 INK4B gene was analyzed with restriction endonucleases combined with PCR technique The expression level of mRNA and protein was detected by RT PCR technique and Western blot Results All detected leukemia cells showed a hypermethylated promoter region of p15 INK4B gene with complete or partial loss of expression of mRNA and protein Either CdR or SB induced weak expression of p15 INK4B with a dose dependent effect Combined low dose of CdR(0 5 μmol/L) and SB(0 5 mmol/L) increased the expression level of p15 INK4B significantly The synergistic effect of the two drugs was significantly stronger than that of any drug of high dose alone Conclusion Hypermethylated and silenced p15 INK4B gene could be re expressed by synergy of CdR and SB更多还原