PHI调控Molt-4细胞组蛋白甲基化诱导p15基因去甲基化后再表达  

Phenylhexyl Isothiocyanate Induces Gene p15 Re-expression by Regulating Histone Methylation and DNA Demethylation in Molt-4 Cells

作　　者：马旭东[1] 黄轶群[1] 蒋少红[1] 郑瑞玑[1] 

机构地区：[1]福建医科大学附属漳州市医院血液科,福建漳州363000

出　　处：《中国实验血液学杂志》2010年第3期583-587,共5页Journal of Experimental Hematology

基　　金：卫生部研究基金福建卫生教育联合攻关计划项目(编号wkj2008-2-55);福建医科大学科学研究发展专项基金计划资助项目(编号FZS08018);漳州市科学研究发展计划基金资助项目(编号Z07014)

摘　　要：本研究探讨异硫氰酸苯己酯(PHI)对急性T淋巴细胞性白血病Molt-4细胞组蛋白H3K4、H3K9甲基化,及p15基因的去甲基化的调控作用及诱导沉默基因重新表达的机制。用Western blot方法检测PHI作用后Molt-4细胞的组蛋白H3K4、H3K9甲基化状态及P15蛋白的表达的变化;应用甲基化特异性聚合酶链反应(MSP)检测PHI作用前后Molt-4细胞株p15基因甲基化状态的变化;半定量逆转录-聚合酶链反应(RT-PCR)检测Molt-4细胞经过PHI处理后p15基因的mRNA的表达变化。结果表明,PHI作用于Molt-4细胞可增强组蛋白H3K4甲基化表达,抑制组蛋白H3K9甲基化表达,并呈浓度及时间依赖性;PHI作用于Molt-4细胞5天后,p15基因的甲基化程度减弱,p15基因的异常高甲基化现象被逆转,沉默的p15基因重新表达;p15 mRNA、P15蛋白表达增加,并呈浓度依赖性。结论:PHI可能通过特异性调节组蛋白H3K4、H3K9甲基化水平,引起染色体空间结构发生变化,使p15基因的CPG岛去甲基化,从而诱导p15基因重新表达。This study was aimed to investigate the regulatory effect of phenylhexyl isothiocyanate （PHI） on methylation of histone H3K4, H3K9 and demethylation of p15 gene in acute leukemia cell line Molt-4, and to explore the possible mechanism inducing rexpression of silent gene. The methylation status of histone H3K4, H3K9 and the expression of P15 protein in the Molt-4 cells treated with PHI were detected by Western blot; the methylation status of p15 gene in the Molt-4 cells before and after treatment with PHI was determined by methylation specific polymerase chain reaction （MSP）; the expression level of p15 gene mRNA in Molt-4 cells treated with PHI was assayed by semiquantitative reverse transcription-PCR. The results indicated that the PHI could increase methylation of histone H3K4 and decrease methylation of histone H3K9 in concentration-and time-dependent manners. After treatment of Molt-4 cells with PHI for 5 days, the methylation of p15 gene was reduced, the significant hypermethylation of p15 gene was reversed, the silenced p15 gene re-expressed; the expressions of p15 mRNA and P15 protein were enhanced in concentration-dependent manner. It is concluded that probably through specifically regulating the methylation level of histone H3K4 and H3K9, the PHI causes the changes of chromosome space structure and results in the demethylation of CPG island in p15 gene, thereby induces the re-expression of p15 gene which was silenced.

关 键 词：异硫氰酸苯己酯 MOLT-4细胞 组蛋白去乙酰化酶抑制剂 组蛋白甲基化 p15基因去甲基化 

分 类 号：R733.1[医药卫生—肿瘤] R979.1[医药卫生—临床医学]