Comparative Promoter Methylation Analysis of p53 Target Genes in Urogenital Cancers

Introduction: The methylation status of selected new p53 target genes in bladder, kidney and testicular cancer was investigated to find similarities in methylation frequency and individual levels of methylation with regard to the histopathological classification and biological behavior. Material and Methods: The patients’ samples were evaluated using real-time methylation-specific PCR (MSP) investigating the methylation status of APAF-1, CASP-8, DAPK-1 and IGFBP-3. Included were 103 patients with bladder cancer, 85 patients with kidney cancer and 42 patients with testicular cancer in non-advanced and advanced tumor stages. Non-cancerous tissue from 49 patients was used as control. Results: Methylation frequency was highest for APAF-1 (88–100%) and DAPK-1 (17–86%) through all the different tumor types. Highest APAF-1 and DAPK-1 levels of methylation were found in the advanced tumor groups. In normal tissue, methylation frequency and methylation levels were significantly lower. APAF-1 methylation was also frequent in normal testicular tissue but to lower methylation levels as in cancerous tissue. APAF-1 and IGFBP-3 methylation levels were significantly higher in recurrent superficial and muscle-invasive bladder tumors. Methylation levels of APAF-1 also correlated with higher risk for recurrent metastatic disease in non-advanced tumors of the kidney. Conclusions: The methylation profile observed demonstrates a substantial role of the APAF-1 gene in tumorigenesis. The different methylation pattern in testicular cancer might represent a different methylotype of this cancer. The extent of promoter methylation may be a promising target for application as a tumor marker in urogenital cancer disease.

[1]  M. Ebinger,et al.  Promoter methylation pattern of caspase-8, P16INK4A, MGMT, TIMP-3, and E-cadherin in medulloblastoma , 2008, Pathology & Oncology Research.

[2]  J. Köllermann,et al.  Promoter Hypermethylation Profile of Kidney Cancer with New Proapoptotic p53 Target Genes and Clinical Implications , 2006, Clinical Cancer Research.

[3]  J. Richie.  Frequent Promoter Hypermethylation of the O6???Methylguanine???DNA Methyltransferase (MGMT) Gene in Testicular Cancer , 2006 .

[4]  Peter A. Jones,et al.  Prognostic relevance of methylation markers in patients with non-muscle invasive bladder carcinoma. , 2005, European journal of cancer.

[5]  F. Marshall.  Promoter hypermethylation profile of kidney cancer. , 2005, The Journal of urology.

[6]  N. Breslow,et al.  Prognostic Value of Quantifying Apoptosis Factor Expression in Favorable Histology Wilms Tumors , 2005, Journal of pediatric hematology/oncology.

[7]  R. Uzzo,et al.  Detection of Bladder Cancer in Urine by a Tumor Suppressor Gene Hypermethylation Panel , 2004, Clinical Cancer Research.

[8]  Shahrokh F. Shariat,et al.  C: IGF-I l IGFBP-3 l ☆ , 2003 .

[9]  L. Jia,et al.  Role of DNA methylation in the suppression of Apaf-1 protein in human leukaemia , 2003, Oncogene.

[10]  T. Wheeler,et al.  Correlation of preoperative levels of IGF-I and IGFBP-3 with pathologic parameters and clinical outcome in patients with bladder cancer. , 2003, Urology.

[11]  R. Lothe,et al.  Frequent promoter hypermethylation of the O6-Methylguanine-DNA Methyltransferase (MGMT) gene in testicular cancer , 2002, Oncogene.

[12]  M. Kuwano,et al.  The association of death-associated protein kinase hypermethylation with early recurrence in superficial bladder cancers. , 2002, Cancer research.

[13]  M. Toyota,et al.  DNA methylation and histone deacetylation associated with silencing DAP kinase gene expression in colorectal and gastric cancers , 2002, British Journal of Cancer.

[14]  K. Aozasa,et al.  [Hypermethylation of DAP-kinase gene CpG Island in malignant lymphoma with B-cell phenotype]. , 2001, Rinsho byori. The Japanese journal of clinical pathology.

[15]  S. Srinivasula,et al.  Apaf-1 protein deficiency confers resistance to cytochrome c-dependent apoptosis in human leukemic cells. , 2001, Blood.

[16]  J. Herman,et al.  A gene hypermethylation profile of human cancer. , 2001, Cancer research.

[17]  W. Gerald,et al.  Inactivation of the apoptosis effector Apaf-1 in malignant melanoma , 2001, Nature.

[18]  J. Herman,et al.  Hypermethylation of the DAP-kinase CpG island is a common alteration in B-cell malignancies. , 1999, Blood.

[19]  K. Bhalla,et al.  Advances in Brief Overexpression of Apaf-1 Promotes Apoptosis of Untreated and Paclitaxel-or Etoposide-treated HL-60 Cells , 2006 .

[20]  S. Srinivasula,et al.  Autoactivation of procaspase-9 by Apaf-1-mediated oligomerization. , 1998, Molecular cell.

[21]  K. Huebner,et al.  Assignment of DAP1 and DAPK--genes that positively mediate programmed cell death triggered by IFN-gamma--to chromosome regions 5p12.2 and 9q34.1, respectively. , 1995, Genomics.

[22]  J. Brooks,et al.  Cytidine methylation of regulatory sequences near the pi-class glutathione S-transferase gene accompanies human prostatic carcinogenesis. , 1994, Proceedings of the National Academy of Sciences of the United States of America.