DLC-1基因甲基化在骨髓增生异常综合征中的临床意义及地西他滨对DLC-1基因表达的影响
Clinical significance of hypermethylation of DLC-1 gene in myelodysplastic syndrome patients and effects of decitabine on DLC-1 gene expression
目的 检测肝癌缺失基因(DLC-1)在骨髓增生异常综合征(MDS)患者中的异常甲基化情况及异常甲基化对MDS患者DLC-1基因的表达影响;探讨DLC-1基因甲基化对MDS的临床意义及MDS的表观遗传学药物地西他滨对DLC-1基因的表达影响.方法 选取福建医科大学附属协和医院血液内科2013至2015年收治的43例MDS患者,采用甲基特异性PCR法(MSP)定性检测MDS患者DIC基因甲基化状态,荧光实时定量PCR(RTFQ-PCR)检测MDS患者DLC-1基因的表达情况;根据MDS的世界卫生组织(WHO)预后积分系统(WPSS积分)标准及危险度分组将MDS患者分为极低危、低危、中危、高危、极高危5组(n=0、8、7、18、10),探讨DLC-1基因甲基化对MDS患者的临床意义.采用重亚硫酸盐测序PCR(BSP)法定量检测地西他滨作用前、后MDS患者DLC-1基因的甲基化状态,RTFQ-PCR检测MDS患者用药前、后DLC-1基因基因mRNA的表达变化.结果 MDS患者DLC-1基因的甲基化比例为55.16% (22/43),DLC-1基因甲基化阳性患者骨髓DLC-1基因mRNA的表达(0.32 ±0.06)显著低于DLC-1基因甲基化阴性患者(0.91 ±0.11) (P=0.008).对于MDS患者,危险度分层中、高危患者的DLC-1基因甲基化率为21/35,明显高于低危患者的1/8(P =0.006).8例采用含地西他滨治疗方案(地西他滨20 mg/m2,共5d,28 d为1疗程,4疗程以上)的患者治疗前、后的DLC基因甲基化率分别为57.50%±5.11%和14.13%±2.07%(P=0.010),DLC-1基因mRNA表达分别为0.28±0.06和0.67 ±0.08(P =0.015),差异均有统计学意义.结论 DLC-1基因甲基化在MDS患者中较为常见,且可能与不良预后相关;表观遗传学治疗药物地西他滨可逆转MDS患者DLC-1基因异常甲基化状态,使DLC-1基因表达上调,恢复其活性,可能是地西他滨治疗MDS患者的机制之一.
更多Objective To detect the methylation status of DLC-1 gene in the patients with myelodysplastic syndrome(MDS),the effect of abnormal methylation of DLC-1 gene on the expression of DLC-1 gene,the clinical significance of methylation of DLC-1 gene in MDS patients,and the effect of decitabine on DLC-1 gene expression.Methods A total of 43 MDS patients were treated in Fujian Medical University Union Hospital from 2013 to 2015.Methylation status of DLC-1 gene in MDS patients were detected by the methylation specific PCR(MSP).The expression of DLC-1 gene mRNA was determined with real-time fluorescence quantitative PCR(RTFQ-PCR).MDS patients were divided into 5 groups (very lowrisk,low-risk,intermediate-risk,high-risk and very high-risk,n =0,8,7,18,10) according to WPSS classification.And the clinical significance of methylation of DLC-1 gene in patients with MDS were investigated.In order to investigate the change in gene methylation and expression of DLC-1 gene after treatment with decitabine,methylation statuses of DLC-1 gene in MDS patients before and after be treated with decitabine were detected by the bisulfite sequencing PCR (BSP).The expressions of DLC-1 gene mRNA of these patients were determined with RTFQ-PCR.Results Hypermethylation of CpG island of DLC-1 gene was observed in 55.16% (22/43) MDS patients.The expressions of DLC-1 gene mRNA in methylation positive patients were significantly lower than that in methylation negative patients (0.32 ± 0.06 vs 0.91 ±0.11) (P =0.008).For MDS patients,the DLC-1 methylation rate of intermediate-and high-risk patient was 21/35,which was significantly higher than that of low-risk patient (1/8,P =0.006).The methylation status of DLC-1 gene were monitored in 8 patients before and after treatment with the decitabine (decitabine 20 mg/m2,dl-d5/d28,more than 4 courses),the methylation rate of DLC-1 gene dropped from 57.50% ± 5.11% to 14.13% ± 2.07% after treatment (P =0.010).The expression of DLC-1 gene increased after treatment with decitabine (0.67 ± 0.08 vs 0.28 ± 0.06,P =0.015).Conclusions Methylation of DLC-1 gene is common in MDS patients and may be associated with poor prognosis.Decitabine may activate the expression of DLC-1 gene by demethylation,which may be one of the mechanisms for the treatment of patients with MDS.
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