Methylation of HOXA9 and ISL1 Predicts Patient Outcome in High-Grade Non-Invasive Bladder Cancer.
Methylation of HOXA9 and ISL1 Predicts Patient Outcome in High-Grade Non-Invasive Bladder Cancer.
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DOI：
10.1371/journal.pone.0137003
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Farrell WE
中科院分区：
文献类型：
--
作者：
Kitchen MO;Bryan RT;Haworth KE;Emes RD;Luscombe C;Gommersall L;Cheng KK;Zeegers MP;James ND;Devall AJ;Fryer AA;Farrell WE
不适当的 DNA 甲基化常常与人类肿瘤的发展相关，在特定情况下，还与临床结果相关。先前关于低/中级非肌层浸润性膀胱癌 (NMIBC) 中 DNA 甲基化的报道表明，DNA 甲基化的特定模式可能具有作为诊断或预后生物标志物的作用。鉴于高级别（HG）NMIBC 的侵袭性和临床不可预测性，以及目前首选治疗方案（芽孢杆菌：卡介苗）的短缺，新型甲基化分析可能类似地揭示疾病结果的生物标志物，从而对患者进行风险分层并指导初始诊断时的临床管理。在 36 个初始表现的高级别 NMIBC、12 个低/中级别 NMIBC 和 3 个正常膀胱对照的原发肿瘤组织中测定了启动子相关的 CpG 岛甲基化。根据先前的报告和/或低/中级别 NMIBC 的预后效用，选择基因 HOXA9、ISL1、NKX6-2、SPAG6、ZIC1 和 ZNF154 进行研究。通过亚硫酸氢钠转化的 DNA 的焦磷酸测序测定甲基化，然后使用 RT-qPCR 与基因表达相关联。甲基化还与肿瘤行为相关，包括肿瘤复发和进展为肌肉浸润性膀胱癌或转移。 ISL1 基因的启动子相关岛在复发性和进展性高级别肿瘤中比非复发性肿瘤更频繁地甲基化（60.0% vs. 18.2%，p = 0.008）。与非复发性肿瘤相比，ISL1 和 HOXA9 在复发性和进展性肿瘤中表现出显着更高的平均甲基化（分别为 43.3% vs. 20.9%，p = 0.016 和 34.5% vs 17.6%，p = 0.017）。 HG-NMIBC 中并发的 ISL1/HOXA9 甲基化可靠地预测一年内肿瘤复发和进展（阳性预测值 91.7%），并且与疾病特异性死亡率 (DSM) 相关。在这项研究中，我们报告了高级别 NMIBC 临床亚型之间甲基化的差异和相似性。我们报告了甲基化生物标志物在初次诊断时预测诊断一年内肿瘤复发和进展的潜在能力。我们发现，尽管高级别 NMIBC 的临床病程和异质性不可预测，但特定的生物标志物可以可靠地预测疾病结果，因此可能有助于指导患者治疗。需要进一步的研究，包括在更大的患者队列中进行验证，以确认甲基化生物标志物在高级别 NMIBC 中的临床效用。
Inappropriate DNA methylation is frequently associated with human tumour development, and in specific cases, is associated with clinical outcomes. Previous reports of DNA methylation in low/intermediate grade non-muscle invasive bladder cancer (NMIBC) have suggested that specific patterns of DNA methylation may have a role as diagnostic or prognostic biomarkers. In view of the aggressive and clinically unpredictable nature of high-grade (HG) NMIBC, and the current shortage of the preferred treatment option (Bacillus:Calmette-Guerin), novel methylation analyses may similarly reveal biomarkers of disease outcome that could risk-stratify patients and guide clinical management at initial diagnosis. Promoter-associated CpG island methylation was determined in primary tumour tissue of 36 initial presentation high-grade NMIBCs, 12 low/intermediate-grade NMIBCs and 3 normal bladder controls. The genes HOXA9, ISL1, NKX6-2, SPAG6, ZIC1 and ZNF154 were selected for investigation on the basis of previous reports and/or prognostic utility in low/intermediate-grade NMIBC. Methylation was determined by Pyrosequencing of sodium-bisulphite converted DNA, and then correlated with gene expression using RT-qPCR. Methylation was additionally correlated with tumour behaviour, including tumour recurrence and progression to muscle invasive bladder cancer or metastases. The ISL1 genes’ promoter-associated island was more frequently methylated in recurrent and progressive high-grade tumours than their non-recurrent counterparts (60.0% vs. 18.2%, p = 0.008). ISL1 and HOXA9 showed significantly higher mean methylation in recurrent and progressive tumours compared to non-recurrent tumours (43.3% vs. 20.9%, p = 0.016 and 34.5% vs 17.6%, p = 0.017, respectively). Concurrent ISL1/HOXA9 methylation in HG-NMIBC reliably predicted tumour recurrence and progression within one year (Positive Predictive Value 91.7%), and was associated with disease-specific mortality (DSM). In this study we report methylation differences and similarities between clinical sub-types of high-grade NMIBC. We report the potential ability of methylation biomarkers, at initial diagnosis, to predict tumour recurrence and progression within one year of diagnosis. We found that specific biomarkers reliably predict disease outcome and therefore may help guide patient treatment despite the unpredictable clinical course and heterogeneity of high-grade NMIBC. Further investigation is required, including validation in a larger patient cohort, to confirm the clinical utility of methylation biomarkers in high-grade NMIBC.