Qin Chao, Chen Jianchun, Li Jie, Ju Xiaobing, Zhang Shaobo, Cao Qiang, Han Zhijian, Li Pu, Shao Peifei, Wang Meilin, Zhang Zhengdong, Gu Min, Zhang Wei, Yin Changjun
State Key Laboratory of Reproductive Medicine, Department of Urology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, Jiangsu 210029, China and Cancer Center of Nanjing Medical University, Department of Molecular and Genetic Toxicology, Nanjing Medical University, 140 Hanzhong Road, Nanjing 210029, China.
Cancer Center of Nanjing Medical University, Department of Molecular and Genetic Toxicology, Nanjing Medical University, 140 Hanzhong Road, Nanjing 210029, China.
Mutagenesis. 2014 Nov;29(6):419-25. doi: 10.1093/mutage/geu046. Epub 2014 Sep 18.
Angiogenesis is fundamentally important to the pathogenesis of clear cell renal cell carcinoma (ccRCC). We investigated the association between variations of genes related to angiogenesis and the risk of ccRCC. In a case-control study of 859 ccRCC patients and 1004 cancer-free subjects, we genotyped 24 potentially functional single nucleotide polymorphisms (SNPs) in seven angiogenesis-related genes (HIF1A, EPAS1, VEGFA, VEGFR1, VEGFR2, VEGFR3 and PDGFRB) using the TaqMan or Snapshot method. Unconditional logistic regression, adjusted for potential confounding factors, was used to assess the risk associations. The functionality of selected SNPs was assessed by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) and luciferase reporter gene assays. We found two SNPs (VEGFA rs2010963 and VEGFR3 rs448012) that were significantly associated with increased risk of ccRCC, after adjusting for multiple comparisons [rs2010963 CC/GC cf. GG: false discovery rate (FDR) = 0.048, odds ratio (OR) = 1.36, 95% confidence interval (95% CI) = 1.12-1.66; rs448012 CC/GC cf. GG: FDR = 0.048, OR = 1.38, 95% CI =1.13-1.69]. Real-time quantitative PCR revealed that the variant genotypes of rs2010963, but not rs448012, were associated with increased gene expression in normal tissues of ccRCC patients (CC/GC cf. GG: P = 0.036). The luciferase reporter assay showed that the rs2010963 C allele significantly increased luciferase activity over that of the rs2010963 G allele. Our results indicate that VEGFA rs2010963 and VEGFR3 rs448012 are associated with risk of ccRCC. Furthermore, rs2010963 is a functional SNP that may affect ccRCC susceptibility by modulating endogenous VEGFA expression.
血管生成在透明细胞肾细胞癌（ccRCC）的发病机制中至关重要。我们研究了血管生成相关基因变异与ccRCC风险之间的关联。在一项针对859例ccRCC患者和1004名无癌受试者的病例对照研究中，我们使用TaqMan或Snapshot方法对7个血管生成相关基因（HIF1A、EPAS1、VEGFA、VEGFR1、VEGFR2、VEGFR3和PDGFRB）中的24个潜在功能性单核苷酸多态性（SNP）进行了基因分型。采用针对潜在混杂因素进行调整的无条件逻辑回归来评估风险关联。通过实时定量逆转录聚合酶链反应（RT-PCR）和荧光素酶报告基因检测评估所选SNP的功能。在进行多重比较校正后，我们发现两个SNP（VEGFA rs2010963和VEGFR3 rs448012）与ccRCC风险增加显著相关[rs2010963 CC/GC对比GG：错误发现率（FDR）=0.048，优势比（OR）=1.36，95%置信区间（95%CI）=1.12 - 1.66；rs448012 CC/GC对比GG：FDR = 0.048，OR = 1.38，95%CI =1.13 - 1.69]。实时定量PCR显示，rs2010963的变异基因型而非rs448012的变异基因型与ccRCC患者正常组织中基因表达增加相关（CC/GC对比GG：P = 0.036）。荧光素酶报告基因检测表明，rs2010963的C等位基因比rs2010963的G等位基因显著增加了荧光素酶活性。我们的结果表明，VEGFA rs2010963和VEGFR3 rs448012与ccRCC风险相关。此外，rs2010963是一个功能性SNP，可能通过调节内源性VEGFA表达影响ccRCC易感性。
