Tumor Specific Epigenetic Silencing of Corticotropin Releasing Hormone -Binding Protein in Renal Cell Carcinoma: Association of Hypermethylation and Metastasis.
The relevance of Corticotropin Releasing Hormone (CRH)-system in human malignancies is a question of growing interest. Here we investigated hypermethylation and epigenetic silencing of the CRH-Binding Protein (CRHBP) gene in clear cell renal cell cancer (ccRCC). Relative methylation of the CRHBP CpG island (CGI) was determined in 17 tumor cell lines as well as 86 ccRCC samples and 66 paired normal tissues using pyrosequencing and quantitative methylation specific PCR of bisulfite converted DNA. Results were statistically compared with relative mRNA expression levels of CRHBP and clinicopathological parameters of patients. Re-expression of CRHBP following 5-aza-2´-deoxycytidine treatment was investigated by quantitative mRNA expression analysis. Real-time impedance analysis was applied for analysis of invasiveness of renal tumor cells following si-RNA knockdown of CRHBP expression or ectopic expression of CRHBP. We found the CRHBP CGI to be frequently methylated in tumor cell lines of renal, prostatic, and bladder cancer. Comparison of methylation in normal and paired renal cancer tissue specimens revealed hypermethylation of the CRHBP CGI in tumors (p<1*10-12). DNA methylation and decreased mRNA expression were correlated (R = 0.83, p<1*10-12). Tumor cell lines showed 5-aza-2´-deoxycytidine dependent reduction of methylation and re-expression of CRHBP was associated with altered cellular invasiveness of renal cancer cells in real-time impedance invasion assays. Hypermethylation and inverse relationship with mRNA expression were validated in silico using the TCGA network data. We describe for the first time tumor specific epigenetic silencing of CRHBP and statistical association with aggressive tumors thus suggesting the CRH system to contribute to the development of kidney cancer.
Department of Urology and Urological Oncology, Hannover Medical School, Hannover, Germany.
Hossein
H
Tezval
Department of Urology and Urological Oncology, Hannover Medical School, Hannover, Germany.
Natalia
N
Dubrowinskaja
Department of Urology and Urological Oncology, Hannover Medical School, Hannover, Germany.
Inga
I
Peters
Department of Urology and Urological Oncology, Hannover Medical School, Hannover, Germany.
Christel
C
Reese
Department of Molecular Biology, Hannover Medical School, Hannover, Germany.
Katrin
K
Serth
Department of Urology and Urological Oncology, Hannover Medical School, Hannover, Germany.
Faranaz
F
Atschekzei
Department of Urology, Eberhard Karls University of Tuebingen, Tuebingen, Germany.
Jörg
J
Hennenlotter
Department of Urology, Eberhard Karls University of Tuebingen, Tuebingen, Germany.
Arnulf
A
Stenzl
Department of Urology and Urological Oncology, Hannover Medical School, Hannover, Germany.
Markus A
MA
Kuczyk
Department of Urology and Urological Oncology, Hannover Medical School, Hannover, Germany.
Jürgen
J
Serth
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002292:Carcinoma, Renal Cell / Q000235:genetics* / Q000473:pathology; D002352:Carrier Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D019175:DNA Methylation / Q000235:genetics*; D044127:Epigenesis, Genetic; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D008297:Male; D008875:Middle Aged; D009362:Neoplasm Metastasis; D012333:RNA, Messenger / Q000235:genetics
D002352:Carrier Proteins; D012333:RNA, Messenger; C067133:corticotropin releasing factor-binding protein
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