Misawa Yuki, Misawa Kiyoshi, Kawasaki Hideya, Imai Atsushi, Mochizuki Daiki, Ishikawa Ryuji, Endo Shiori, Mima Masato, Kanazawa Takeharu, Iwashita Toshihide, Mineta Hiroyuki
1 Department of Otorhinolaryngology/Head and Neck Surgery, Hamamatsu University School of Medicine, Hamamatsu, Japan.
2 Department of Regenerative and Infectious Pathology, Hamamatsu University School of Medicine, Hamamatsu, Japan.
Tumour Biol. 2017 Jul;39(7):1010428317711657. doi: 10.1177/1010428317711657.
The aim of this study was to determine the methylation status of the genes encoding the vascular endothelial growth factor receptors and to evaluate the usefulness of VEGFR methylation as a prognostic indicator in head and neck squamous cell carcinoma. VEGFR messenger RNA expression and promoter methylation were examined in a panel of cell lines via quantitative reverse transcription and methylation-specific polymerase chain reaction, respectively. Promoter methylation was compared with clinical characteristics in 128 head and neck squamous cell carcinoma samples. The normalized methylation values for the VEGFR1, VEGFR2 and VEGFR3 promoters tended to be higher in the tumour cell lines than in normal tonsil samples, whereas amounts of VEGFR1, VEGFR2 and VEGFR3 messenger RNA were significantly higher. Methylation of the VEGFR1 promoter (p = 0.003; 66/128 head and neck squamous cell carcinoma samples, 52%) and VEGFR3 promoter (p = 0.043; 53/128 head and neck squamous cell carcinoma samples, 41%) significantly correlated with recurrence, whereas methylation of the VEGFR2 promoter significantly correlated with lymph node metastasis (p = 0.046; 47/128 head and neck squamous cell carcinoma samples, 37%). Concurrent methylation of the VEGFR1 and VEGFR3 promoters significantly correlated with reduced disease-free survival (log-rank test, p = 0.009). In a multivariate logistic regression analysis, methylation of the VEGFR1, VEGFR3 and both the VEGFR1 and VEGFR3 promoters independently predicted recurrence (odds ratios and 95% confidence intervals: 3.19, 1.51-6.75 (p = 0.002); 2.24, 1.06-4.76 (p = 0.035); and 2.56, 1.09-6.05 (p = 0.032), respectively). Methylation of the VEGFR promoters predicts poor prognosis in head and neck squamous cell carcinoma patients.
本研究的目的是确定血管内皮生长因子受体编码基因的甲基化状态，并评估VEGFR甲基化作为头颈部鳞状细胞癌预后指标的实用性。分别通过定量逆转录和甲基化特异性聚合酶链反应检测了一组细胞系中的VEGFR信使核糖核酸表达和启动子甲基化。将启动子甲基化与128例头颈部鳞状细胞癌样本的临床特征进行了比较。VEGFR1、VEGFR2和VEGFR3启动子的标准化甲基化值在肿瘤细胞系中往往高于正常扁桃体样本，而VEGFR1、VEGFR2和VEGFR3信使核糖核酸的量则显著更高。VEGFR1启动子甲基化（p = 0.003；128例头颈部鳞状细胞癌样本中的66例，52%）和VEGFR3启动子甲基化（p = 0.043；128例头颈部鳞状细胞癌样本中的53例，41%）与复发显著相关，而VEGFR2启动子甲基化与淋巴结转移显著相关（p = 0.046；128例头颈部鳞状细胞癌样本中的47例，37%）。VEGFR1和VEGFR3启动子的同时甲基化与无病生存期缩短显著相关（对数秩检验，p = 0.009）。在多变量逻辑回归分析中，VEGFR1、VEGFR3以及VEGFR1和VEGFR3启动子的甲基化独立预测复发（优势比和95%置信区间：分别为3.19，1.51 - 6.75（p = 0.002）；2.24，1.06 - 4.76（p = 0.035）；以及2.56，1.09 - 6.05（p = 0.032））。VEGFR启动子甲基化预示着头颈部鳞状细胞癌患者的预后不良。
