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• W2607452364 abstract "AACR Annual Meeting-- Apr 12-16, 2008; San Diego, CA4281 Background: GSTP1 has been proposed to be a caretaker gene, protecting cells against genome damage. It has been shown that the GSTP1 protein is absent in prostate cancer (CaP) and high grade prostatic intraepithelial neoplasia (HGPIN). The absence of GSTP1 protein in CaP and HGPIN is related to hypermethylation of the GSTP1 CpG island. Previously we reported that while normal epithelium from cancer patients does not contain methylated GSTP1 , 6% of atrophy lesions, 68.8% of HGPIN lesions and 90.9% of CaP lesions harbor methylated GSTP1 alleles (Nakayama et al. Am J Pathol. 2003; 923-33). Moreover, density of methylation, as well as presence of methylation was likely to be associated with prostate carcinogenesis. However, the pattern of methylation of individual alleles has not been investigated previously. We used bisulfite sequencing to investigate methylation status of 39 CpG sites within the GSTP1 promoter. Design: Radical prostatectomy specimens were selected from men at The Johns Hopkins Hospital. Three 6-µm sections were cut for laser capture microdissection. The DNA was isolated with standard phenol extraction. Genomic DNA was bisulfite converted, followed by nested PCR. PCR products were sub-cloned and plasmid DNA from 10 clones was sequenced and analyzed. DNA from the prostate cancer cell line LNCaP wasused as a positive control for GSTP1 methylation, normal humanWBC DNA (Novagen) was used as a negative control. Results: The methylation levels of 39 CpG sites of the GSTP1 promoter region (-254¼+81) were determined. 163 clones from normal epithelium (18 regions), 225 clones from atrophy (25 regions), 142 clones from HGPIN (16 regions), and 153 clones from CaP (18 regions) were successfully sequenced. In general, normal epithelium and atrophic areas were either completely unmethylated or only slightly methylated--33 of 6057 sites (0.5%) were methylated in normal, 70 of 8475 sites (0.8%) were methylated in atrophy. By contrast, HGPIN lesions showed increased methylation, and the pattern tended to show partial methylation of individual clones-- 1166 of 5399 sites (22%) of HGPIN CpG sites were methylated. CaP showed a stepwise increase, usually having much more dense methylation than HGPIN--3285 of 5819 (57%) sites were methylated in CaP. 6 proximally located CpG sites (-4¼+38) were considered hot spots in HGPIN clones since among 53 moderately (7-19 out of 39) or highly (20-39) methylated clones, 50 had at least 5 methylated CpG sites in this area. Conclusion: The results support the hypothesis that methylation of individual CpG sites in the island gradually increases during progression of the neoplastic process. Since HGPIN lesions were clearly intermediate between normal and cancer, these results further support the concept that HGPIN lesions are indeed part of the stepwise molecular progression in prostate carcinogenesis." @default.
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• W2607452364 date "2008-05-01" @default.
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• W2607452364 title "Mapping of DNA methylation status of GSTP1 CpG island in human prostate tissue" @default.
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