论文部分内容阅读
目的:探讨细胞信号转导抑制因子1(suppressor of cytokine signaling1,SOCS1)基因超甲基化在经典型慢性骨髓增殖性肿瘤(myelo-proliferative neoplasms,MPNs)中的临床作用及其机制。方法:采用甲基化特异性PCR法检测MPNs患者骨髓标本中SOCS1基因的超甲基化发生情况。应用粒细胞集落刺激因子(granulocyte colony stimulating factor,G-CSF)化学诱导MPNs细胞生长不同时间后,实时定量PCR和蛋白质印迹法检测SOCS1mRNA及其蛋白表达情况。另外,加入去甲基化试剂2-deoxyazacytidin培养MPNs细胞不同时间后,实时定量PCR法检测SOCS1mRNA表达情况。结果:100例经典型慢性MPNs患者中有27例(27.0%)存在SOCS1基因超甲基化。G-CSF化学诱导处理后的MPNs细胞中,SOCS1基因超甲基化组与未甲基化组相比,其SOCS1mRNA表达量均明显减少(均P<0.05);MPNs患者中SOCS1基因超甲基化组的SOCS1蛋白表达量也减少,但与健康对照组相比差异尚无统计学意义(P>0.05)。另外,相对于G-CSF处理后的超甲基化组,加入去甲基化试剂后SOCS1mRNA表达量明显升高(P<0.05)。结论:MPNs患者中存在SOCS1基因超甲基化,且CpG岛超甲基化导致其基因表达水平降低,去甲基化后,其基因表达水平升高。提示SOCS1基因超甲基化可能和MPNs发病有关,去甲基化可能是一种潜在的治疗MPNs的新策略。
Objective: To investigate the clinical significance and mechanism of hypermethylation of suppressor of cytokine signaling1 (SOCS1) gene in classic chronic myeloid proliferative neoplasms (MPNs). Methods: Methylation-specific PCR was used to detect the hypermethylation of SOCS1 gene in bone marrow samples of patients with MPNs. The growth of MPNs was induced by granulocyte colony stimulating factor (G-CSF) at different time points. The mRNA and protein expression of SOCS1 were detected by real-time quantitative PCR and Western blotting. In addition, after adding demethylation reagent 2-deoxyazacytidin to culture MPNs cells for different time, the expression of SOCS1 mRNA was detected by real-time quantitative PCR. RESULTS: SOCS1 gene hypermethylation was present in 27 of 27 (27.0%) patients with classical chronic MPNs. SOCS1mRNA expression in SOCS1 hypermethylated group was significantly lower than that in unmethylated group (all P <0.05) in MPNs treated with G-CSF. In MPNs, SOCS1 gene hypermethylation The expression of SOCS1 protein also decreased, but the difference was not statistically significant compared with the healthy control group (P> 0.05). In addition, compared with G-CSF-treated hypermethylation group, SOCS1mRNA expression was significantly increased (P <0.05) after addition of demethylation reagent. CONCLUSIONS: SOCS1 gene hypermethylation exists in patients with MPNs, and hypermethylation of CpG island leads to the decrease of gene expression level. After demethylation, the gene expression level of SOCS1 gene is increased. These results suggest that hypermethylation of SOCS1 may be related to the pathogenesis of MPNs. Demethylation may be a potential new strategy for the treatment of MPNs.