Promoter Hypermethylation Profile of Kidney Cancer with New Proapoptotic p53 Target Genes and Clinical Implications

Purpose: Risk stratification of renal cell carcinoma is based on the histopathologic classification. Promoter hypermethylation as a mechanism of gene inactivation in renal cell carcinoma has been shown for only a small number of genes. We examined the usefulness of quantitative methylation analysis with a new set of p53 target genes for determining the clinical outcome and aggressiveness of the tumor disease. Experimental Design: The genes selected were APAF-1, CASPASE-8, DAPK-1, IGFBP-3, and PML. The tissue samples analyzed were taken from tumor specimens obtained from 90 consecutive patients with clear cell renal carcinoma and from 20 normal kidney specimens. Quantitative methylation analysis of CpG sites in the promoter region was done by methylation-specific real-time PCR and the normalized index of methylation (NIM) was determined for each sample. Results: Hypermethylation of the promoter region was common for APAF-1 (97%) and DAPK-1 (41%) but not for IGFBP-3 (3%), PML (3%), or CASP-8 (0%). The tumor patients had a median follow-up of 55 months. A correlation was found between the methylation level of APAF-1 and tumor size and nodal status, but not for tumor stage, grade, and age of patient. Kaplan-Meier analysis was able to identify patients with a higher risk of recurrence and tumor-related death by using APAF-1 (≥56% NIM) and DAPK-1 (≥10% NIM) methylation levels. In multivariate analysis, APAF-1 and DAPK-1 methylation levels were independent prognostic markers for metastatic disease and death from renal cell carcinoma. Conclusions: Our findings indicate that promoter hypermethylation of APAF-1 and DAPK-1 is a marker of aggressive renal cell carcinoma and provides independent prognostic information on disease outcome.

[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]  Xin-yang Wang,et al.  Overexpression of PML induced apoptosis in bladder cancer cell by caspase dependent pathway. , 2006, Cancer letters.

[3]  C. Suschek,et al.  Expression of death-associated protein kinase during tumour progression of human renal cell carcinomas: hypermethylation-independent mechanisms of inactivation. , 2006, European journal of cancer.

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

[5]  P. Carroll,et al.  Methylation of the γ-Catenin Gene is Associated With Poor Prognosis of Renal Cell Carcinoma , 2005 .

[6]  P. Carroll,et al.  Methylation of the g -Catenin Gene Is Associated With Poor Prognosis of Renal Cell Carcinoma , 2005 .

[7]  C. Rogers,et al.  Molecular profiling and classification of sporadic renal cell carcinoma by quantitative methylation analysis. , 2006, Clinical cancer research : an official journal of the American Association for Cancer Research.

[8]  P. Pandolfi,et al.  Mutations of the PML tumor suppressor gene in acute promyelocytic leukemia. , 2004, Blood.

[9]  P. Pandolfi,et al.  PML is a direct p53 target that modulates p53 effector functions. , 2004, Molecular cell.

[10]  A. Jemal,et al.  Cancer Statistics, 2004 , 2004, CA: a cancer journal for clinicians.

[11]  L. Schmidt,et al.  The genetic basis of renal cell carcinoma. , 2003, The Urologic clinics of North America.

[12]  A. Chan,et al.  Frequent hypermethylation of promoter region of RASSF1A in tumor tissues and voided urine of urinary bladder cancer patients , 2003, International journal of cancer.

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

[14]  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.

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

[16]  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.

[17]  Peter A. Jones,et al.  The fundamental role of epigenetic events in cancer , 2002, Nature Reviews Genetics.

[18]  R. Sylvester,et al.  Radical nephrectomy plus interferon-alfa-based immunotherapy compared with interferon alfa alone in metastatic renal-cell carcinoma: a randomised trial , 2001, The Lancet.

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

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

[21]  A. Eggert,et al.  Resistance to TRAIL-induced apoptosis in primitive neuroectodermal brain tumor cells correlates with a loss of caspase-8 expression , 2000, Oncogene.

[22]  C Béroud,et al.  p53 Website and analysis of p53 gene mutations in human cancer: Forging a link between epidemiology and carcinogenesis , 2000, Human mutation.

[23]  Gustavo Droguett,et al.  DAP kinase activates a p19ARF/p53-mediated apoptotic checkpoint to suppress oncogenic transformation , 2000, Nature Cell Biology.

[24]  P. Cohen,et al.  Insulin-like Growth Factor (IGF)-binding Protein-3 Induces Apoptosis and Mediates the Effects of Transforming Growth Factor-β1 on Programmed Cell Death through a p53- and IGF-independent Mechanism* , 1997, The Journal of Biological Chemistry.

[25]  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.

[26]  A. Kimchi,et al.  Identification of a novel serine/threonine kinase and a novel 15-kD protein as potential mediators of the gamma interferon-induced cell death. , 1995, Genes & development.

[27]  J. Brooks,et al.  Mutations of the VHL tumour suppressor gene in renal carcinoma , 1994, Nature Genetics.