Nickerson Michael L, Jaeger Erich, Shi Yangu, Durocher Jeffrey A, Mahurkar Sunil, Zaridze David, Matveev Vsevolod, Janout Vladimir, Kollarova Hellena, Bencko Vladimir, Navratilova Marie, Szeszenia-Dabrowska Neonilia, Mates Dana, Mukeria Anush, Holcatova Ivana, Schmidt Laura S, Toro Jorge R, Karami Sara, Hung Rayjean, Gerard Gary F, Linehan W Marston, Merino Maria, Zbar Berton, Boffetta Paolo, Brennan Paul, Rothman Nathaniel, Chow Wong-Ho, Waldman Frederic M, Moore Lee E
Transgenomic, Gaithersburg, Maryland, USA.
Clin Cancer Res. 2008 Aug 1;14(15):4726-34. doi: 10.1158/1078-0432.CCR-07-4921.
To provide a comprehensive, thorough analysis of somatic mutation and promoter hypermethylation of the von Hippel-Lindau (VHL) gene in the cancer genome, unique to clear cell renal cancer (ccRCC). Identify relationships between the prevalence of VHL gene alterations and alteration subtypes with patient and tumor characteristics.
As part of a large kidney cancer case-control study conducted in Central Europe, we analyzed VHL mutations and promoter methylation in 205 well-characterized, histologically confirmed patient tumor biopsies using a combination of sensitive, high-throughput methods (endonuclease scanning and Sanger sequencing) and analysis of 11 CpG sites in the VHL promoter.
We identified mutations in 82.4% of cases, the highest VHL gene mutation prevalence reported to date. Analysis of 11 VHL promoter CpG sites revealed that 8.3% of tumors were hypermethylated and all were mutation negative. In total, 91% of ccRCCs exhibited alteration of the gene through genetic or epigenetic mechanisms. Analysis of patient and tumor characteristics revealed that certain mutation subtypes were significantly associated with Fuhrman nuclear grade, metastasis, node positivity, and self-reported family history of RCC.
Detection of VHL gene alterations using these accurate, sensitive, and practical methods provides evidence that the vast majority of histologically confirmed ccRCC tumors possess genetic or epigenetic alteration of the VHL gene and support the hypothesis that VHL alteration is an early event in ccRCC carcinogenesis. These findings also indicate that VHL molecular subtypes can provide a sensitive marker of tumor heterogeneity among histologically similar ccRCC cases for etiologic, prognostic, and translational studies.
对透明细胞肾细胞癌（ccRCC）特有的癌症基因组中冯·希佩尔-林道（VHL）基因的体细胞突变和启动子高甲基化进行全面、深入的分析。确定VHL基因改变的发生率和改变亚型与患者及肿瘤特征之间的关系。
作为在中欧进行的一项大型肾癌病例对照研究的一部分，我们使用灵敏的高通量方法（内切酶扫描和桑格测序）以及对VHL启动子中11个CpG位点的分析，对205例特征明确、经组织学确诊的患者肿瘤活检样本中的VHL突变和启动子甲基化进行了分析。
我们在82.4%的病例中发现了突变，这是迄今为止报道的VHL基因突变发生率最高的。对11个VHL启动子CpG位点的分析显示，8.3%的肿瘤发生了高甲基化，且所有这些肿瘤均为突变阴性。总体而言，91%的ccRCC通过遗传或表观遗传机制表现出该基因的改变。对患者和肿瘤特征的分析表明，某些突变亚型与福尔曼核分级、转移、淋巴结阳性以及自我报告的肾癌家族史显著相关。
使用这些准确、灵敏且实用的方法检测VHL基因改变提供了证据，表明绝大多数经组织学确诊的ccRCC肿瘤具有VHL基因的遗传或表观遗传改变，并支持VHL改变是ccRCC致癌过程中早期事件的假说。这些发现还表明，VHL分子亚型可为病因学、预后和转化研究中组织学相似的ccRCC病例间的肿瘤异质性提供灵敏的标志物。
