肾母细胞瘤和成人肾细胞癌的多基因甲基化分析

Multigene methylation analysis of Wilms' tumour and adult renal cell carcinoma.

作者信息

Morris Mark R, Hesson Luke B, Wagner Kate J, Morgan Neil V, Astuti Dewi, Lees Robert D, Cooper Wendy N, Lee JouAnn, Gentle Dean, Macdonald Fiona, Kishida Takeshi, Grundy Richard, Yao Masahiro, Latif Farida, Maher Eamonn R

机构信息

Section of Medical and Molecular Genetics, Department of Paediatrics and Child Health, University of Birmingham, Birmingham B15 2TT, UK.

出版信息

Oncogene. 2003 Oct 2;22(43):6794-801. doi: 10.1038/sj.onc.1206914.

DOI:10.1038/sj.onc.1206914

PMID:14555992

Abstract

To investigate the role of epigenetic gene silencing in the pathogenesis of Wilms' tumour and renal cell carcinoma (RCC), we determined their methylation profile using a candidate gene approach. Thus, 40 Wilms' tumours and up to 49 adult RCC were analysed by methylation-specific PCR for promoter methylation at CASP8, CDH1, CDH13, DAPK, MGMT, NORE1A, p14ARF and RARB2 in primary Wilms' tumours and CASP8, CDH1, CDH13, CRBP1, DAPK, MGMT, MT1G, NORE1A, p16INK4a, SDHB and RARB2 in primary RCC. Both tumour sample sets had previously been analysed for RASSF1A promoter methylation, and p16INK4a methylation results were also available for the Wilms' tumour samples. Wilms' tumours demonstrated a high incidence of methylation at CASP8 (43%) and MGMT (30%), intermediate frequencies at NORE1A (15%), p14ARF (15%), p16INK4a (10%), DAPK (11%) and CRBP1 (9%), but promoter methylation was rare or absent at RARB2 (0%), CDH13 (0%) and CDH1 (3%). No association was detected between methylation of RASSF1A, CASP8 or MGMT in individual tumours. The frequency of MGMT methylation was higher in stage 1 and 2 tumours (50%) than in stage 3 and 4 tumours (17%) but this did not reach statistical significance (P=0.06). RCC were most frequently methylated at DAPK (24%), MT1G (20%), NORE1A (19%), CDH1 (16%) and MGMT (9%) and not or rarely at SDHB (4%), RARB2 (0%), p16INK4a (0%) and CDH13 (3%). There were no associations between methylation of RASSF1A, DAPK and CDH1 in individual tumours. Papillary RCC demonstrated a higher frequency of DAPK methylation (43%) than clear cell tumours (19%) (P=0.14). We have demonstrated that de novo promoter methylation is frequent in Wilms' tumour and RCC, and these data enable methylation profiles to be constructed for each tumour type. Thus, combining our results with data published previously, it appears that promoter methylation occurs frequently (> or =20% of primary tumours) at CASP8, SLIT2 and RASSF1A in Wilms' tumour and at RASSF1A, TIMP3, DAPK, SLIT2, MT1G and GSTP1 in RCC.

摘要

为了研究表观遗传基因沉默在肾母细胞瘤和肾细胞癌（RCC）发病机制中的作用，我们采用候选基因方法确定了它们的甲基化谱。因此，通过甲基化特异性PCR分析了40例肾母细胞瘤和多达49例成人RCC，检测原发性肾母细胞瘤中CASP8、CDH1、CDH13、DAPK、MGMT、NORE1A、p14ARF和RARB2的启动子甲基化情况，以及原发性RCC中CASP8、CDH1、CDH13、CRBP1、DAPK、MGMT、MT1G、NORE1A、p16INK4a、SDHB和RARB2的启动子甲基化情况。这两组肿瘤样本之前都已分析过RASSF1A启动子甲基化情况，并且肾母细胞瘤样本的p16INK4a甲基化结果也已获得。肾母细胞瘤中，CASP8（43%）和MGMT（30%）甲基化发生率较高，NORE1A（15%）、p14ARF（15%）、p16INK4a（10%）、DAPK（11%）和CRBP1（9%）的甲基化频率中等，但RARB2（0%）、CDH13（0%）和CDH1（3%）的启动子甲基化罕见或不存在。在单个肿瘤中，未检测到RASSF1A、CASP8或MGMT甲基化之间的关联。MGMT甲基化在1期和2期肿瘤（50%）中的频率高于3期和4期肿瘤（17%），但未达到统计学意义（P = 0.06）。RCC中最常发生甲基化的是DAPK（24%）、MT1G（20%）、NORE1A（19%）、CDH1（16%）和MGMT（9%），而SDHB（4%）、RARB2（0%）、p16INK4a（0%）和CDH13（3%）甲基化不存在或罕见。在单个肿瘤中，RASSF1A、DAPK和CDH1甲基化之间无关联。乳头状RCC中DAPK甲基化频率（43%）高于透明细胞肿瘤（19%）（P = 0.14）。我们已经证明，原发性启动子甲基化在肾母细胞瘤和RCC中很常见，这些数据使我们能够为每种肿瘤类型构建甲基化谱。因此，将我们的结果与之前发表的数据相结合，似乎在肾母细胞瘤中，CASP8、SLIT2和RASSF1A启动子甲基化频繁发生（≥20%的原发性肿瘤），在RCC中，RASSF1A、TIMP3、DAPK、SLIT2、MT1G和GSTP1启动子甲基化频繁发生。