Zhao Youshan, Fei Chengming, Zhang Xi, Zhang Yao, Guo Juan, Gu Shucheng, Li Xiao, Chang Chunkang
Department of Hematology, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiaotong University, 200233, Shanghai, China.
Tumour Biol. 2013 Feb;34(1):165-72. doi: 10.1007/s13277-012-0524-y. Epub 2012 Sep 28.
The identification of epigenetically inactivated tumor suppressor genes may shed additional light on the pathogenesis of myelodysplastic syndromes (MDS) and lead to the identification of potential therapeutic targets for demethylating agents. In this study, the methylation status of the p73 gene promoter was analyzed by methylation-specific PCR (MS-PCR) in bone marrow (BM) samples from 126 adult patients with de novo MDS. The results of the MS-PCR were confirmed by bisulfite sequencing. In addition, we analyzed p73 expression using real-time PCR. The apoptosis of BM cells was examined by flow cytometry. The methylation of the p73 gene was observed in 36.5 % of cases. There were strong correlations between p73 methylation and the marrow blast levels (p = 0.037) and the WHO classification (p = 0.016). The frequency of p73 methylation was significantly correlated with the International Prognostic Scoring System subgroup ( r = 0.904, p < 0.001). Moreover, a decrease in the transcription of p73 was accompanied by methylation (p = 0.032). Although the level of apoptosis in the BM samples of the methylated group was not significantly lower than that in the unmethylated group (p = 0.094), decitabine treatment restored p73 expression and increased the level of cytarabine (ara-C)-induced apoptosis in vitro. The median survival time of patients with p73 methylation was shorter than that for patients without p73 methylation (15 vs. > 33 months, respectively, p = 0.002). A multivariate analysis also indicated that the p73 methylation status was the independent factor that impacted overall survival. In conclusion, p73 methylation is common in patients with MDS and is associated with poor prognosis. Our results provide further evidence for the involvement of epigenetic dysregulation in the pathogenesis of MDS.
表观遗传失活的肿瘤抑制基因的鉴定可能为骨髓增生异常综合征(MDS)的发病机制提供更多线索,并有助于确定去甲基化药物的潜在治疗靶点。在本研究中,采用甲基化特异性PCR(MS-PCR)分析了126例成年初诊MDS患者骨髓(BM)样本中p73基因启动子的甲基化状态。MS-PCR结果通过亚硫酸氢盐测序进行了验证。此外,我们使用实时PCR分析了p73的表达。通过流式细胞术检测BM细胞的凋亡情况。在36.5%的病例中观察到p73基因的甲基化。p73甲基化与骨髓原始细胞水平(p = 0.037)和世界卫生组织(WHO)分类(p = 0.016)之间存在强相关性。p73甲基化频率与国际预后评分系统亚组显著相关(r = 0.904,p < 0.001)。此外,p73转录的降低与甲基化相伴(p = 0.032)。虽然甲基化组BM样本中的凋亡水平并不显著低于未甲基化组(p = 0.094),但地西他滨治疗可恢复p73表达并在体外增加阿糖胞苷(ara-C)诱导的凋亡水平。p73甲基化患者的中位生存时间短于无p73甲基化的患者(分别为15个月和> 33个月,p = 0.002)。多因素分析还表明,p73甲基化状态是影响总生存的独立因素。总之,p73甲基化在MDS患者中常见且与预后不良相关。我们的结果为表观遗传失调参与MDS发病机制提供了进一步证据。