Hypomethylation of WNT5A, CRIP1 and S100P in prostate cancer.
Oligoarray analysis of a matched pair of prostate cancer and normal cell lines derived from the same radical prostatectomy specimen identified 113 candidate hypomethylated genes that were overexpressed in the cancer cells and contained CpG islands. Hypomethylation of wingless-related MMTV integration site 5A (WNT5A), S100 calcium-binding protein P (S100P) and cysteine-rich protein 1(CRIP1) was confirmed in the cancer cells by bisulfite sequencing. Treatment of the corresponding normal prostate epithelial cells 1542-NPTX with the DNA methyltransferase inhibitor 5-Aza-2'-deoxycytidine (5-aza-CdR) induced higher levels of mRNA expression and partial loss of methylation on these genes. Primary prostate cancers were tested using methylation-specific polymerase chain reaction. WNT5A was hypomethylated in 11/17 (65%) tumors, S100P in 8/16 (50%) and CRIP1 in 13/20 (65%). Bisulfite sequencing of a section of the 5' untranslated region (UTR) of WNT5A revealed that three CpG sites (15, 24 and 35) were consistently methylated (93%) in the normal cell line and normal tissues, but not in the prostate cancer cell line and eight primary prostate cancers. Multiple putative binding sites for the transcription factors SP1 and AP-2 were found adjacent to CpG sites 15 and 24. A putative c-Myb binding site was located within the CpG site 35. Anti-c-Myb antibody co-precipitation with WNT5A was methylation-sensitive in 1542-NPTX cells. It is likely that an epigenetic mechanism regulates WNT5A expression in prostate cancer.
Wang
Q
Q
Prostate Cancer Research Centre, Institute of Urology and Nephrology, University College London, and Department of Histopathology, University College London Hospitals Trust, UK.
Williamson
M
M
Bott
S
S
Brookman-Amissah
N
N
Freeman
A
A
Nariculam
J
J
Hubank
M J F
MJ
Ahmed
A
A
Masters
J R
JR
D001665:Binding Sites; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D018899:CpG Islands; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D060588:LIM Domain Proteins; D008297:Male; D009363:Neoplasm Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011471:Prostatic Neoplasms / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016329:Sp1 Transcription Factor / Q000378:metabolism; D014407:Tumor Cells, Cultured; D051153:Wnt Proteins / Q000378:metabolism*; D000071818:Wnt-5a Protein
C486181:CRIP1 protein, human; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D060588:LIM Domain Proteins; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins; C103003:S100P protein, human; D016329:Sp1 Transcription Factor; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein