This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Jerónimo, C. Articles by Sidransky, D. Search for Related Content PubMed PubMed Citation Articles by Jerónimo, C. Articles by Sidransky, D.

I105V Polymorphism and Promoter Methylation of the GSTP1 Gene in Prostate Adenocarcinoma¹

Departments of Pathology [C. J., G. V., R. H., C. S., C. L.], Genetics [C. J.], Urology [J. O.], and Epidemiology [M. J. B.], Portuguese Institute of Oncology, 4200–072 Porto, Portugal; and Department of Otolaryngology-Head and Neck Surgery, Head and Neck Cancer Research Division, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205 [D. S.]

	Abstract

Top Abstract Introduction Materials and Methods Results Discussion References

 
The GSTP1 gene encodes for an enzyme, glutathione S-transferase (GST),involved in detoxification of carcinogens. An aminoacid substitution (I105V) in GSTP1 produces a variant enzyme with lower activity and less capability of effective detoxification. This variant GSTP*B allele has been associated with a propensity to develop several neoplasms. Because GSTP1 promoter hypermethylation and inactivation of GST expression is a frequent alteration in prostate carcinoma, we hypothesized that this somatic epigenetic modification could obviate any reduced enzyme activity caused by the germ-line polymorphism. We tested for the GSTP1 genotype in a population of prostate cancer patients, and in a control group composed of patients with benign prostatic hyperplasia (BPH) and healthy blood donors. Tissue samples from the 105 prostate cancer cases (105 adenocarcinomas and 34 prostatic intraepithelial neoplasia lesions), and from 43 BPH patients were tested for GSTP1 hypermethylation by methylation-specific PCR. GST protein expression was assessed by immunohistochemistry. No significant effect on prostate cancer risk was detectable for GSTP1 genotype compared with the control population (odds ratio, 1.02; 95% confidence interval, 0.59–1.75). Moreover, no association was found between this genotype and tumor or BPH methylation status. Patients with unmethylated carcinomas did not disclose significant differences in genotypic distribution compared with the control population. In adenocarcinoma, a strong association (P < 0.00001) between GSTP1 promoter hypermethylation and loss of GST expression was observed; however, this trend was not retained in prostatic intraepithelial neoplasia or BPH lesions. Although the GSTP1 polymorphism is not associated with altered susceptibility to prostate cancer, somatic promoter hypermethylation is an effective, but not the only, cause of decreased GST function.

	Introduction

Top Abstract Introduction Materials and Methods Results Discussion References

 
Prostate adenocarcinoma is the most frequently diagnosed cancer among men in the Western world, and the second leading cause of cancer death among men in the United States (1) . Etiologically, prostate cancer is a multifactorial disease in which several environmental and genetic factors may be involved, yet little is known about the interaction between these factors (2) . Moreover, the role of epigenetic phenomena, such as DNA de novo methylation, in the modulation of gene expression, has become a major focus of investigation in the field of prostate cancer research (3) .

GSTP1, located at 11q13, belongs to a supergene family of enzymes, the GSTs,³ involved in the detoxification of electrophilic compounds (such as carcinogens and cytotoxic drugs) by glutathione conjugation (4 , 5) . In addition, these enzymes are believed to play a role in the protection of DNA from oxidative damage (6) . GSTP1 has a polymorphic site at codon 105 (exon 5), where an adenosine-to-guanosine (A-G) transition causes an Iso-to-Val substitution (I105V), giving rise to the GSTP1*B allele (4, 5, 6, 7) . Moreover, recent studies have found that individuals with the valine allele display a significantly lower enzyme activity and less effective capability of detoxification (8) . Hence, an association between GSTP*B allele and lung, bladder, and testicular neoplasms has been reported (6 , 7) . A significant decrease in the frequency of GSTP1*A has been reported in prostate cancer patients compared with controls (7) , but these results have been challenged by other studies, using larger series of patients (9, 10, 11, 12) .

Over the last few years, several studies revealed that GSTP1 (usually expressed in normal human epithelial tissues, including prostate) could be somatically inactivated by hypermethylation of the promoter region (13, 14, 15) . This alteration, which is often associated with a loss of GST expression, is the most common event (90%) described thus far in prostate adenocarcinoma (15 , 16) . Furthermore, GSTP1 inactivation may lead to an increased cell vulnerability to oxidative DNA damage and to the accumulation of DNA base adducts, which can make a tumor cell move to acquire other relevant genetic alterations in prostatic carcinogenesis (17 , 18) . Thus, we hypothesized that this somatic epigenetic modification could obviate any effect on the differential enzyme activity caused by allelic variants and could perhaps explain the conflicting reports on the effect of the GSTP1 polymorphism in prostate cancer risk.

We first investigated the association between the I105V GSTP1 polymorphism and the risk for developing prostate cancer, to investigate a primary genotypic effect on prostate cancer susceptibility. Then, we searched for a possible association between this polymorphism and de novo methylation in a relatively large series of early-stage (clinically localized) prostate cancer patients (19 , 20) . Finally, immunohistochemical analysis was done to determine whether GSTP1 hypermethylation affects gene expression.

	Materials and Methods

Top Abstract Introduction Materials and Methods Results Discussion References

 
GSTP1 Polymorphism
Blood Samples and DNA Extraction.
For this study, two populations of male subjects were enrolled at the Portuguese Institute of Oncology, Porto, Portugal. One population consisted of 105 patients with histologically confirmed adenocarcinoma. The control population comprised 43 patients with BPH, and 98 healthy male volunteer blood donors from the same institution. Blood was collected from all of the individuals, and genomic DNA was extracted from fresh peripheral leukocytes as described previously (21) . Briefly, DNA was digested overnight at 48°C in 1% SDS/proteinase K (0.5 mg/ml), extracted by phenol-chloroform, and ethanol precipitated.

GSTP1 Genotype Analysis.
The exon 5 polymorphic site in GSTP1 locus (Ile-105Val) was detected by restriction fragment length polymorphism of PCR-amplified fragments.

The primers used were: P105 forward, 5'-ACC CCA GGG CTC TAT GGG AA-3', and P105 reverse 5'-TGA GGG CAC AAG AAG CCC CT-3' (7) . PCR reactions were carried out in a 30-µl volume containing about 50 ng of genomic DNA template, 200 µM each dNTP, 200 ng each primer, 1.5 mM MgCl₂, 1x PCR buffer [50 mM KCl, 10 mM Tris-HCl (pH 8.3)] and 1 unit Taq DNA polymerase (Promega, Southampton, United Kingdom). After an initial denaturation step of 10 min at 95°C, the samples were processed through 30 temperature cycles of 30 s at 94°C, 30 s at 55°C, and 30 s at 72°C. A final extension step of 72°C for 10 min was performed. The 176-bp PCR products (20 µl) were digested for 2 h at 37°C with 2 units of Alw26I (Fermentas Inc, Vilnius, Lithuania). The detection of the different alleles was carried out by horizontal ethidium bromide 4% agarose gel electrophoresis, along with a 100-bp DNA ladder.

Methylation Analysis
Patients and Tissue Sample Collection.
All of the patients from the prostate cancer group harbored clinically localized prostate adenocarcinoma [T_1c, according to the TNM staging system (22) ], and were consecutively diagnosed and treated with radical prostatectomy. The 43 patients with BPH were submitted to TURP and harbored no histological evidence of malignancy. Two pathologists (R. H., C. L.) reviewed all of the histological slides, and each tumor was graded according to the Gleason grading system (23) . Fresh tissue, snap-frozen in isopentane and stored at -80°C, or paraffin-embedded prostatic tissue was collected from each surgical specimen. Sections were cut for the identification of areas of high-grade PIN and adenocarcinoma (radical prostatectomy specimens), and BPH (TURP tissue). These areas were then carefully microdissected from 12-µm-thick sections for enrichment of PIN, adenocarcinoma, and hyperplastic tissue. On average, 50 sections for each area with enrichment (>70%) in neoplastic cells were used for DNA extraction of PIN or cancer. Paraffin-embedded tissue was similarly microdissected but was placed in xylene for 3 h at 48°C to remove the paraffin. DNA was extracted using the method described by Ahrendt et al. (21) .

Bisulfite Treatment.
Sodium bisulfite conversion of 2 µg of genomic DNA was performed by a modification of a previously described method (24) . Briefly, NaOH was added to denature DNA (final concentration, 0.2 M) and incubated for 20 min at 50°C. A volume of 500 µl freshly made bisulfite solution [2.5 M sodium metabisulfite and 125 mM hydroquinone (pH = 5.0)] was added to each sample, and incubation was continued at 50°C for 3 h in the dark. Modified DNA was purified using the Wizard DNA purification resin according to the manufacturer (Promega Corp., Madison, WI) and was eluted in 45 µl of water at 80°C. After treatment with NaOH (final concentration, 0.3 M) for 10 min at 37°C, isolation was continued with 75 µl of 7.5 M ammonium acetate, followed by an incubation step of 5 min at room temperature. Finally, the modified DNA was precipitated by adding 2.5 volumes of 100% ethanol and 2 µl of glycogen (5 mg/ml). The pellet was washed with 70% ethanol, dried, and eluted in 30 µl of 5 mM Tris (pH 8.0).

MSP Analysis.
For PCR amplification, 2 µl of bisulfite-modified DNA was added in a final volume of 25 µl of PCR mix containing 1x PCR buffer [16.6 mM ammonium sulfate/67 mM Tris (pH 8.8) and 6.7 mM MgCl₂/10 mM 2-mercaptoethanol], dNTPs (each at 1.25 mM), 1 unit Platinum Taq DNA polymerase (Life Technologies, Inc., Rockville, MD) and primers (300 ng each per reaction). Primer sequences for either methylated or modified unmethylated GSTP1 have been described previously (25) . MSP was carried out using the following conditions: 1 cycle at 95°C for 1 min; 35 cycles of 1 min 95°C, 1 min 62°C, and 1 min 72°C, and a final extension for 5 min at 70°C. In each performed PCR, treated DNA extracted from two prostate cancer cell lines, the LNCaP and Du145 were used as positive and negative controls, respectively. The PCR products were directly loaded onto a nondenaturing 6% polyacrylamide gel, stained with ethidium bromide, and visualized under UV illumination.

Immunohistochemical Analysis
Four-µm sections were cut and placed in aminopropyltriethoxysilane (Sigma) coated slides. After dewaxing the sections, endogenous peroxidase activity was inhibited with freshly prepared 0.5% hydrogen peroxide in distilled water for 20 min. Then, they were processed in a 600-W microwave oven at maximum power, three times for 2 min, each time in citrate buffer (pH 6). Immunostaining was performed using an immunoperoxidase method according to the manufacturer’s instructions (Vectastain ABC kit; Vector Laboratories, Burlingame CA). The incubation of the primary anti-GST antibody (clone 3 BD; Transduction Laboratories, Lexington, KY) was performed overnight at 4°C, at a dilution of 1:250 in 1% BSA in PBS. Sections were developed with a peroxidase substrate solution (0.05% 3,3-diaminobenzidine tetrahydrochloride, 0.01% H₂O₂ in PBS), counterstained with hematoxylin, dehydrated, and mounted. Appropriate positive controls were used for each antibody, and negative controls consisted of the replacement of the primary antibody for 1% BSA in PBS.

Assessment of GST expression was performed by light microscopy at x400. The presence or absence of immunostaining was evaluated in morphologically normal areas, PIN lesions, and tumor, as well as in BPH samples.

Statistical Analysis
The odds ratio and 95% confidence intervals were calculated as a measure of the association between GSTP1 genotype and the risk of development of prostate cancer. Associations between GSTP1 genotype and methylation status, as well as any correlation between GSTP1 methylation and GST expression were examined using the ² test, and Fisher’s exact test, when appropriate. Analyses were conducted using a computer-assisted program, Epi Info, version 6 (Centers for Disease Control and Prevention, Atlanta, GA). Statistical significance was reached when P < 0.05.

	Results

Top Abstract Introduction Materials and Methods Results Discussion References

 
Population Characteristics and Distribution of GSTP1 Genotypes.
One hundred five early-stage prostate cancer patients and 141 controls (43 patients with BPH and 98 healthy male volunteers) were recruited for this study. The median age was 63 years (range, 48–74) and 56 years (range, 45–82), for prostate cancer cases and controls, respectively. The age distributions differed significantly between the two groups (P < 0.00001).

Table 1 depicts the frequency distribution of each GSTP1 genotype (Fig. 1) among the prostate cancer cases and controls considered for this study, and no statistically significant difference was found (P = 0.32). Concerning the control group, the frequency distribution of GSTP1*A/*A, GSTP1*A/*B, and GSTP1*B/*B among the BPH patients was 16 (37.2%), 24 (55.8%), and 3 (7%), respectively. For the blood donor population, the frequency distribution for the same genotypes was, respectively, 45 (45.9%), 43 (43.9%), and 10 (10.2%). Statistical analysis did not disclose a significant difference in genotype distribution between the BPH patients and blood donors (P = 0.42). No significant effect on prostate cancer risk was detectable for the GSTP1 genotype (odds ratio, 1.02; 95% confidence interval, 0.59–1.75) compared with the control population.

View larger version (41K): [in this window] [in a new window]   Fig. 1. PCR-restriction fragment length polymorphism analysis of the GSTP1 Ile-105Val polymorphism. The consensus sequence corresponding to the GSTP1*A allele was not cut, whereas the Val sequence corresponding to GSTP1*B was cleaved to yield two fragments (91 bp and 85 bp). Homozygous wild-type (GSTP1*A/*A), heterozygote (GSTP1*A/*B), and homozygous mutant (GSTP1*B/*B) cases are depicted in Lanes 2–4, respectively. Lane 1, the 100-bp DNA ladder; Lane 5, the water control.

View larger version (23K): [in this window] [in a new window]   Fig. 2. Illustrative example of MSP for GSTP1 promoter region in prostate tissues from nonneoplastic areas (MN), and clinically localized prostate adenocarcinoma (T) of patients 54 and 90. Lanes U and M correspond to unmethylated (97 bp) and methylated (93 bp) reactions, respectively. In each case, DNA from normal lymphocytes was used as negative control for methylation (U+), DNA from LNCaP cell line was used as positive control for methylation (M+), and water was used as negative PCR control (H2O). On the right side, the 100-bp DNA ladder.

Immunohistochemical Analysis.
In normal and hyperplastic tissue, GST immunoreactivity was always present in basal cells (Fig. 3A) . This staining was mainly cytoplasmic but nuclear staining was also a frequent finding. Luminal secretory cells displayed much weaker and inconstant staining than basal cells. No difference in immunostaining was observed between BPH cases with or without GSTP1 hypermethylation.

View larger version (168K): [in this window] [in a new window]   Fig. 3. GST immunoexpression in the prostate. A, a case with GSTP1 promoter methylation and absence of GST expression. on the left side, a normal gland showing strong immunoreactivity in basal cells. B, an adenocarcinoma without GSTP1 promoter methylation and with strong immunoreactivity for GST, both cytoplasmic and nuclear. C, a nonmethylated PIN lesion with no immunoreactivity for GST in the dysplastic columnar cells, although the basal cells are positive. D, a GSTP1-methylated PIN lesion shows immunoreactivity for GST in all cell types (basal and columnar epithelial).

View this table: [in this window] [in a new window]   Table 3 Immunohistochemical findings for GST in prostate cancer patients (adenocarcinoma and PIN lesions) according to GSTP1 methylation status

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion References

In the present study, we found no evidence of a differential risk for prostate adenocarcinoma among men possessing the Iso or Val variants of codon 105 of GSTP1. This result is in accordance with most previously published studies (9, 10, 11) , but is in disagreement with the recent report of Gsur et al. (12) . However, a major limitation of our work and the latter study is the control population. Gsur et al. used 166 age-matched control patients with BPH, and we used both a group of BPH patients and a group of healthy blood donors. In this study, the observed difference in age distribution between the blood donors and the cancer patients could be potentially problematic because of the latency of prostate cancer. The inclusion of a group of BPH patients partially overcomes this problem, and no difference in GSTP1 genotype frequencies between the control group and the prostate cancer patients was noted.

Another potential limitation of our study would be the exclusion of patients with advanced-stage prostate cancer, because these patients are not candidates for radical prostatectomy procedures. In this study, only patients with clinically localized disease, submitted to radical prostatectomy were included, and thus, this group would not reflect the true incident population at risk for prostate cancer. However, because no association between GSTP1 genotype and stage in prostate cancer patients has been found (11) , this potential limitation is obviated.

To the best of our knowledge, this study is the first ever to report the frequency distribution of GSTP1 genotypes in a Portuguese population. Furthermore, it is noteworthy that the genotype frequencies found in our series of patients and controls fall in the expected range for a Western European population (7 , 27 , 28) . Although the control populations in some of these studies included women (27 , 28) , no gender influence in GSTP1 genotype distribution has been reported thus far.

GSTP1 promoter hypermethylation is a frequent alteration in prostate cancer cells and is associated with gene silencing and decreased GST expression (15 , 16) . Thus, we hypothesized that this epigenetic modification could obviate the difference in enzyme activity caused by the I105V polymorphism, unless the polymorphism itself would influence GSTP1 promoter methylation status. MSP analysis of GSTP1 promoter hypermethylation in prostate adenocarcinoma tissue samples obtained from the PA group disclosed a high percentage of methylated tumors (84.9%), which is in accordance with previously published results (13 , 14 , 19 , 20) . Moreover, no association was found between GSTP1 hypermethylation and the GSTP1 genotype. The A–G substitution (I105V polymorphism) occurs at position 1578 (exon 5), and GSTP1 hypermethylation associated with prostate cancer takes place at the gene promoter region (5, 6, 7, 8 , 16) . In fact, exon 5 is enriched in methylated CpG sites even in normal tissue (16) , and, thus, it would be unlikely that an A–G substitution so far downstream would influence methylation in the gene promoter.

If, hypothetically, a low GST activity caused by allelic variation (e.g., I105V polymorphism), were linked to prostate cancer pathogenesis, then the patients with unmethylated tumors would be expected to show a higher frequency of the I105V polymorphism. Our results do not sustain this hypothesis because the genotype distribution of the patients with unmethylated tumors (16 cases) did not differ significantly from the control population. Yet, this finding does not exclude a role for GSTP1 in prostate carcinogenesis because other mechanisms responsible for a decrease in GST activity may exist, as demonstrated by the lack of immunoreactivity for GST in 10 of these 16 unmethylated carcinomas.

To confirm the regulation of GSTP1 promoter hypermethylation in GST expression, we performed an immunohistochemical analysis in the radical prostatectomy and TURP specimens from our patients. The immunohistochemical findings confirm that GSTP1 promoter hypermethylation is related to the loss of GST expression in prostate cancer, because all methylated tumors lacked GST. Previous studies reached the same conclusion (13 , 16) , and similar findings were also reported in breast cancer (25) . However, a novel finding from our study is the lack of GST expression in 10 primary tumors not displaying GSTP1 methylation. This result suggests that alternative mechanisms for GSTP1 transcription inactivation may occur in addition to promoter hypermethylation. Interestingly, some prostate cancer cells displaying GST immunoreactivity have been found to lack enzyme activity (16) . Moreover, loss of expression of GST associated with GSTP1 promoter methylation has been identified in potential precursor lesions such as PIN (15) . Hence, a loss of GST function appears to play a critical role in the early steps of prostate carcinogenesis.

For this reason, we also analyzed 34 PIN lesions from the radical prostatectomy specimens. GSTP1 hypermethylation has been reported in 50–70% of PIN lesions (15 , 20) , and other researchers were unable to detect GST expression in this preneoplastic condition (29) . However, we found immunoreactivity for GST in 7 of 34 cases, 5 of which were methylated at the GSTP1 promoter region. This result may be related to the difference in GSTP1 methylation levels found between PIN and adenocarcinoma (20) . In this respect, it is noteworthy that all of the methylated BPH lesions herein analyzed, expressed GST. and these lesions also displayed a significantly lower level of GSTP1 methylation (20) . These findings favor the existence of a critical level of methylation for the silencing of GSTP1.

Different patterns of GST expression in PIN lesions from the transition and the peripheral zone of the prostate have been previously reported (30) . In the present study only PIN lesions from the peripheral zone were analyzed, but even lesions therein located were found to differentially express GST in that report (30) . In our study, the tissue samples microdissected for methylation analysis and those used for immunohistochemical analysis in adenocarcinoma, PIN, or BPH lesions) were very closely colocated. Thus, it is unlikely that methylation results and immunohistochemical findings in each case reflect different foci of a given type of lesion. Although genetic heterogeneity of PIN lesions is acknowledged (31) , no data concerning variation in GSTP1 promoter methylation status across the same prostate specimen has been reported to date.

Our results confirm that GSTP1 promoter hypermethylation is a highly prevalent event in prostate cancer; it is already present in potential precursor lesions and is tightly linked to GST loss of expression. Moreover, the 1105V GSTP1 polymorphism is not associated with hypermethylation in the promoter region nor related with altered susceptibility to prostate cancer. These findings suggest that common somatic GSTP1 inactivation obviates any major effects from inherited genotypic variants in prostate cancer progression.

	Acknowledgments

 
We acknowledge the support of the Liga Portuguesa Contra o Cancro-Núcleo Regional do Norte in the present study.

	Footnotes

 
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ C. J. is supported by a grant from the Fundação para a Ciência e Tecnologia, Portugal (Program PRAXIS XXI-BD 13398/97). Funding for the study described in this article was partially provided by Virco, Inc. Under a licensing agreement between The Johns Hopkins University and Virco, Dr. Sidransky is entitled to a share of royalty received by the University on sales or products described in this article. The University and Dr. Sidransky own Virco stock, which is subject to certain restrictions under University policy. Dr. Sidransky is a paid consultant to Virco. The terms of this arrangement are being managed by The Johns Hopkins University in accordance with its conflict-of-interest policies.

² To whom requests for reprints should be addressed, at Department of Genetics, Portuguese Institute of Oncology, Rua Dr. António Bernardino de Almeida 4200–072 Porto, Portugal. Phone: 351-22-550-20-11; Fax: 351-22-502-64-89; E–mail: carmenjeronimo@netc.pt.

³ The abbreviations used are: GST, glutathione S-transferase; BPH, benign prostatic hyperplasia; PIN, prostatic intraepithelial neoplasia; TURP, transurethral resection of the prostate; MSP, methylation-specific (PCR).

Received 9/ 7/01; revised 2/22/02; accepted 3/ 4/02.

	References

Top Abstract Introduction Materials and Methods Results Discussion References

 

1. Landis S. H., Murray T., Bolden S., Wingo P. A. Cancer statistics. CA Cancer Clin., 49: 8-31, 1999.[Abstract/Free Full Text]
2. Isaacs W. B., Bova G. B. Prostate cancer Vogelstein B. Kinzler K. W. eds. . The Genetic Basis of Human Cancer, 653-660, McGraw-Hill New York 1998.
3. Rennie P. S., Nelson C. C. Epigenetic mechanisms for progression of prostate cancer. Cancer Metastasis. Rev., 17: 401-409, 1999.[Medline]
4. Zimniak P., Nanduri B., Pikula S., Bandorowicz-Pikula J., Singhal S. S., Srivastava S. K., Awasthi S., Awasthi Y. C. Naturally occurring human glutathione S-transferase GSTP1–1 isoforms with isoleucine and valine in position 104 differ in enzymatic properties. Eur. J. Biochem., 224: 893-899, 1994.[Medline]
5. Henderson C. J., McLaren A. W., Moffat G. J., Bacon E. J., Wolf C. R. -class glutathione S-transferase: regulation and function. Chem. Biol. Interact., 111–112: 69-82, 1998.
6. Ryberg D., Skaug V., Hewer A., Phillips D. H., Harries L. W., Wolf C. R., Ogreid D., Ulvik A., Vu P., Haugen A. Genotypes of glutathione transferase M1 and P1 and their significance for lung DNA adduct levels and cancer risk. Carcinogenesis (Lond.), 18: 1285-1289, 1997.[Abstract/Free Full Text]
7. Harries L. W., Stubbins M. J., Forman D., Howard G. C., Wolf C. R. Identification of genetic polymorphisms at the glutathione S-transferase locus and association with susceptibility to bladder, testicular and prostate cancer. Carcinogenesis (Lond.), 18: 641-644, 1997.[Abstract/Free Full Text]
8. Hu X., Xia H., Srivastava S. K., Pal A., Awasthi Y. C., Zimniak P., Singh S. V. Catalytic efficiencies of allelic variants of human glutathione S-transferase P1–1 toward carcinogenic anti-diol epoxides of benzo[c]phenanthrene and benzo[g]chrysene. Cancer Res., 58: 5340-5343, 1998.[Abstract/Free Full Text]
9. Autrup J. L., Thomassen L. H., Olsen J. H., Wolf H., Autrup H. Glutathione S-transferases as risk factors in prostate cancer. Eur. J. Cancer Prev., 8: 525-532, 1999.[Medline]
10. Wadelius M., Autrup J. L., Stubbins M. J., Andersson S. O., Johansson J. E., Wadelius C., Wolf C. R., Autrup H., Rane A. Polymorphisms in NAT2, CYP2D6, CYP2C19 and GSTP1 and their association with prostate cancer. Pharmacogenetics, 9: 333-340, 1999.[Medline]
11. Shepard T. F., Platz E. A., Kantoff P. W., Nelson W. G., Isaacs W. B., Freije D., Febbo P. G., Stampfer M. J., Giovannucci E. No association between the I105V polymorphism of the glutathione S-transferase P1 gene (GSTP1) and prostate cancer risk: a prospective study. Cancer Epidemiol. Biomark. Prev., 9: 1267-1268, 2000.[Free Full Text]
12. Gsur A., Haidinger G., Hinteregger S., Bernhofer G., Schatzl G., Madersbacher S., Marberger M., Vutuc C., Micksche M. Polymorphisms of glutathione-S-transferase genes (GSTP1, GSTM1 and GSTT1) and prostate-cancer risk. Int. J. Cancer, 95: 152-155, 2001.[Medline]
13. Lee W-H., Morton R. A., Epstein J. I., Brooks J. D., Campbell P. A., Bova G. S., Hsieh W-S., Isaacs W. B., Nelson W. G. Cytidine methylation of regulatory sequences near the pi-class glutathione S-transferase gene accompanies human prostatic carcinogenesis. Proc. Natl. Acad. Sci. USA, 91: 11733-11737, 1994.[Abstract/Free Full Text]
14. Lee W-H., Isaacs W. B., Bova G. S., Nelson W. G. CG island methylation changes near the GSTP1 gene in prostatic carcinoma cells detected using the polymerase chain reaction: a new prostate cancer biomarker. Cancer Epidemiol. Biomark. Prev., 6: 443-450, 1997.[Abstract]
15. Brooks J. D., Weinstein M., Lin X., Sun Y., Pin S. S., Bova G. S., Epstein J. I., Isaacs W. B., Nelson W. G. CG island methylation changes near the GSTP1 gene in prostatic intraepithelial neoplasia. Cancer Epidemiol. Biomark. Prev., 7: 531-536, 1998.[Abstract]
16. Millar D. S., Ow K. K., Paul C. L., Russell P. J., Molloy P. L., Clark S. J. Detailed methylation analysis of the glutathione S-transferase (GSTP1) gene in prostate cancer. Oncogene, 18: 1313-1324, 1999.[Medline]
17. Nelson C. P., Kidd L. C., Sauvageot J., Isaacs W. B., De Marzo A. M., Groopman J. D., Nelson W. G., Kensler T. W. Protection against 2-hydroxyamino-1-methyl-6-phenylimidazo [4,5-b]pyridine cytotoxicity and DNA adduct formation in human prostate by glutathione S-transferase P1. Cancer Res., 61: 103-109, 2001.[Abstract/Free Full Text]
18. Berhane K., Widersten M., Engström Å., Kozarich J. W., Mannervik B. Detoxication of base propenals and other , ß-unsaturated aldehyde products of radical reactions and lipid peroxidation by human glutathione transferases. Proc. Natl. Acad. Sci. USA, 91: 1480-1484, 1994.[Abstract/Free Full Text]
19. Cairns P., Esteller M., Herman J. G., Schoenberg M., Jerónimo C., Sanchez-Cespedes M., Show N-H., Grasso M., Wu L., Westra W. B., Sidransky D. Detection of prostate cancer in urine by GSTP1 hypermethylation. Clin. Cancer Res., 7: 2727-2730, 2001.[Abstract/Free Full Text]
20. Jerónimo C., Usadel H., Henrique R., Oliveira J., Lopes C., Nelson W. G., Sidransky D. Quantitation of GSTP1 hypermethylation distinguishes between non-neoplastic prostatic tissue and organ confined prostate adenocarcinoma. J. Natl. Cancer Inst. (Bethesda), 93: 1747-1752, 2001.[Abstract/Free Full Text]
21. Ahrendt S. A., Chow J. T., Xu L-H, Yang S. C., Eisenberger C. F., Esteller M., Herman J. G., Wi L., Decker P. A., Jen J., Sidransky D. Molecular detection of tumor cells in bronchoalveolar lavage fluid from patients with early stage lung cancer. J. Natl. Cancer Inst. (Bethesda), 91: 332-339, 1999.[Abstract/Free Full Text]
22. Hermanek P., Hutter R. V. P., Sobin L. H., Wagner G., Wittekind C. Prostate Hermanek P. Hutter R. V. P. Sobin L. H. Wagner G. Wittekind C. eds. . Illustrated Guide to the TNM/pTNM Classification of Malignant Tumors, 272-280, Springer-Verlag Heidelberg, Germany 1997.
23. Gleason D. F., Mellinger G. T., Veterans Administration Cooperative Urological Research Group. Prediction of prognosis for prostatic adenocarcinoma by combined histologic grading and clinical staging. J. Urol., 111: 58-64, 1974.[Medline]
24. Olek A., Oswald J., Walter J. A. A modified and improved method of bisulfite based cytosine methylation analysis. Nucleic Acids Res., 24: 5064-5066, 1996.[Abstract/Free Full Text]
25. Esteller M., Corn P. G., Urenal J. M., Gabrielson E., Baylin S. B., Herman J. G. Inactivation of glutathione S-transferase P1 gene by promoter hypermethylation in human neoplasia. Cancer Res., 58: 4515-4518, 1999.[Abstract/Free Full Text]
26. Helzlsouer K. J., Selmin O., Huang H. Y., Strickland P. T., Hoffman S., Alberg A. J., Watson M., Comstock G. W., Bell D. Association between glutathione S-transferase M1, P1, and T1 genetic polymorphisms and development of breast cancer. J. Natl. Cancer Inst. (Bethesda), 90: 512-518, 1998.[Abstract/Free Full Text]
27. Allan J. M., Wild C. P., Rollinson S., Willet E. V., Moorman A. V., Dovey G. J., Roddam P. L., Roman E., Cartwright R. A., Morgan G. Polymorphism in glutathione S-transferase P1 is associated with susceptibility to chemotherapy-induced leukemia. Proc. Natl. Acad. Sci. USA, 98: 11592-7, 2001.[Abstract/Free Full Text]
28. van Lieshout E. M. M., Roelofs H. M. J., Dekker S., Mulder C. J. J., Wobbes, Jansen T. J. B. M. J., Peters W. H. M. Polymorphic expression of the glutathione S-transferase P1 gene and its susceptibility to Barrett’s esophagus and esophageal carcinoma. Cancer Res., 59: 586-589, 1999.[Abstract/Free Full Text]
29. Moskaluk C. A., Duray P. H., Cowan K. H., Linehan M., Merino M. J. Immunohistochemical expression of -class glutathione S-transferase is down-regulated in adenocarcinoma of the prostate. Cancer (Phila.), 79: 1595-1599, 1997.[Medline]
30. Montironi R., Mazzucchelli R., Stramazzotti D., Pomante R., Thompson D., Bartels P. H. Expression of -class glutathione S-transferase: two populations of high grade prostatic intraepithelial neoplasia with different relations to carcinoma. Mol. Pathol., 53: 122-128, 2000.[Abstract/Free Full Text]
31. Bostwick D. G., Shan A., Qian J., Darson M., Maihle N. J., Jenkins R. B., Cheng L. Independent origin of multiple foci of prostatic intraepithelial neoplasia. Cancer (Phila.), 83: 1995-2002, 1998.[Medline]

This article has been cited by other articles:

				R. Henrique, C. Jeronimo, M. R. Teixeira, M. O. Hoque, A. L. Carvalho, I. Pais, F. R. Ribeiro, J. Oliveira, C. Lopes, and D. Sidransky Epigenetic Heterogeneity of High-Grade Prostatic Intraepithelial Neoplasia: Clues for Clonal Progression in Prostate Carcinogenesis Mol. Cancer Res., January 1, 2006; 4(1): 1 - 8. [Abstract] [Full Text] [PDF]

				G. S. Palapattu, S. Sutcliffe, P. J. Bastian, E. A. Platz, A. M. De Marzo, W. B. Isaacs, and W. G. Nelson Prostate carcinogenesis and inflammation: emerging insights Carcinogenesis, July 1, 2005; 26(7): 1170 - 1181. [Abstract] [Full Text] [PDF]

				Q. K.Y. Chan, U.-S. Khoo, H. Y.S. Ngan, C.-Q. Yang, W.-C. Xue, K. Y.K. Chan, P.-M. Chiu, P. P.C. Ip, and A. N.Y. Cheung Single Nucleotide Polymorphism of Pi-Class Glutathione S-Transferase and Susceptibility to Endometrial Carcinoma Clin. Cancer Res., April 15, 2005; 11(8): 2981 - 2985. [Abstract] [Full Text] [PDF]

				L.-C. Li, P. R. Carroll, and R. Dahiya Epigenetic Changes in Prostate Cancer: Implication for Diagnosis and Treatment J Natl Cancer Inst, January 19, 2005; 97(2): 103 - 115. [Abstract] [Full Text] [PDF]

				C. Ntais, A. Polycarpou, and J. P.A. Ioannidis Association of GSTM1, GSTT1, and GSTP1 Gene Polymorphisms with the Risk of Prostate Cancer: A Meta-analysis Cancer Epidemiol. Biomarkers Prev., January 1, 2005; 14(1): 176 - 181. [Abstract] [Full Text] [PDF]

				C Jeronimo, R Henrique, J Oliveira, F Lobo, I Pais, M R Teixeira, and C Lopes Aberrant cellular retinol binding protein 1 (CRBP1) gene expression and promoter methylation in prostate cancer J. Clin. Pathol., August 1, 2004; 57(8): 872 - 876. [Abstract] [Full Text] [PDF]

				J. V. Tricoli, M. Schoenfeldt, and B. A. Conley Detection of Prostate Cancer and Predicting Progression: Current and Future Diagnostic Markers Clin. Cancer Res., June 15, 2004; 10(12): 3943 - 3953. [Abstract] [Full Text] [PDF]

				C. Jeronimo, R. Henrique, M. O. Hoque, F. R. Ribeiro, J. Oliveira, D. Fonseca, M. R. Teixeira, C. Lopes, and D. Sidransky Quantitative RAR{beta}2 Hypermethylation: A Promising Prostate Cancer Marker Clin. Cancer Res., June 15, 2004; 10(12): 4010 - 4014. [Abstract] [Full Text] [PDF]

				Y. Zhu, M. R. Spitz, C. I. Amos, J. Lin, M. B. Schabath, and X. Wu An Evolutionary Perspective on Single-Nucleotide Polymorphism Screening in Molecular Cancer Epidemiology Cancer Res., March 15, 2004; 64(6): 2251 - 2257. [Abstract] [Full Text] [PDF]

				L. Kopelovich, J. A. Crowell, and J. R. Fay The Epigenome as a Target for Cancer Chemoprevention J Natl Cancer Inst, December 3, 2003; 95(23): 1747 - 1757. [Abstract] [Full Text] [PDF]

				M. Nakayama, C. J. Bennett, J. L. Hicks, J. I. Epstein, E. A. Platz, W. G. Nelson, and A. M. De Marzo Hypermethylation of the Human Glutathione S-Transferase-{pi} Gene (GSTP1) CpG Island Is Present in a Subset of Proliferative Inflammatory Atrophy Lesions but Not in Normal or Hyperplastic Epithelium of the Prostate: A Detailed Study Using Laser-Capture Microdissection Am. J. Pathol., September 1, 2003; 163(3): 923 - 933. [Abstract] [Full Text] [PDF]

				M. VERMA, B. K. DUNN, S. ROSS, P. JAIN, W. WANG, R. HAYES, and A. UMAR Early Detection and Risk Assessment Proceedings and Recommendations from the Workshop on Epigenetics in Cancer Prevention Ann. N.Y. Acad. Sci., March 1, 2003; 983(1): 298 - 319. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Jerónimo, C. Articles by Sidransky, D. Search for Related Content PubMed PubMed Citation Articles by Jerónimo, C. Articles by Sidransky, D.