High circulating estrogens and selective expression of ERβ in prostate tumors of Americans: implications for racial disparity of prostate cancer

Zakaria Elmageed

Krzysztof Moroz

Sudesh Srivastav

Zhide Fang

Supplementary Material

Supplementary Data:

Click here to view.

^{Department of Urology,}

^{Department of Pathology, and}

^{Department of Biostatistics and Bioinformatics, Tulane University Health Science Center, New Orleans, LA 70112, USA,}

^{Biostatistics Program, School of Public Health, LSU Health Sciences Center, New Orleans, LA 70112, USA, and}

^{Tulane Cancer Center, Tulane University Health Science Center, New Orleans, LA 70112, USA}

*To whom correspondence should be addressed. Department of Urology, Tulane University School of Medicine, 1430 Tulane Avenue, New Orleans, LA 70112, USA. Tel: +504 988 3836; Fax: +504 988 5059; Email: ude.enalut@deegama

Received 2012 Nov 21; Revised 2013 Apr 12; Accepted 2013 May 5.

Abstract

Although estrogen receptor beta (ERβ) has been implicated in prostate cancer (PCa) progression, its potential role in health disparity of PCa remains elusive. The objective of this study was to examine serum estrogens and prostate tumor ERβ expression and examine their correlation with clinical and pathological parameters in African American (AA) versus Caucasian American (CA) men. The circulating 17β-estradiol (E2) was measured by enzyme immunoassay in blood procured from racially stratified normal subjects and PCa patients. Differential expression profile analysis of ERβ was analyzed by quantitative immunohistochemistry using ethnicity-based tissue microarray encompassing 300 PCa tissue cores. In situ ERβ expression was validated by quantitative reverse transcription–PCR in matched microdissected normal prostate epithelium and tumor cells and datasets extracted from independent cohorts. In comparison with normal age-matched subjects, circulating E2 levels were significantly elevated in all PCa patients. Further analysis demonstrates an increase in blood E2 levels in AA men in both normal and PCa in comparison with age- and stage-matched counterparts of CA decent. Histochemical score analysis reveals intense nuclear immunoreactivity for ERβ in tumor cores of AA men than in CA men. Gene expression analysis in microdissected tumors corroborated the biracial differences in ERβ expression. Gene expression analysis from independent cohort datasets revealed correlation between ERβ expression and PCa progression. However, unlike in CA men, adjusted multivariate analysis showed that ERβ expression correlates with age at diagnosis and low prostate-specific antigen recurrence-free survival in AA men. Taken together, our results suggest that E2-ERβ axis may have potential clinical utility in PCa diagnosis and clinical outcome among AA men.

Abstract

Pearson correlation coefficient (r) is calculated among groups and number in bold indicates significance at P < 0.05.

Pearson correlation coefficient (r) is calculated among groups and P < 0.05 is considered significant (bold numbers).

Click here to view.

Acknowledgements

The authors thank Ms Grace Kelly for support in scoring analysis of TMAs by SlidePath image software (SlidePath Ltd, Dublin, Ireland). The authors also thank Ms Jennifer Cvitanovic from LCRC Biospecimen core for providing human PCa tissue sections and serum samples.

Conflict of Interest Statement: None declared.

Acknowledgements

Glossary

Abbreviations:

AA	African American
AAD	age at diagnosis
BPH	benign prostatic hyperpasia
CA	Caucasian American
CRPC	castration-resistant PCa
E2	17β-estradiol
ERβ	estrogen receptor beta
GS	Gleason score
HS	H-Score, histochemical score
PCa	prostate cancer
PRF	PSA recurrence-free
PSA	prostate-specific antigen
TMA	tissue microarray.

Glossary

References

1. Couse J.F, et al. (1999). Estrogen receptor null mice: what have we learned and where will they lead us?Endocr. Rev., 20, 358–417 [[PubMed]
2. Gustafsson J.A. (2003) What pharmacologists can learn from recent advances in estrogen signalling. Trends Pharmacol. Sci., 24, 479–485 [[PubMed][Google Scholar]
3. Chang E.C, et al. (2006). Impact of estrogen receptor beta on gene networks regulated by estrogen receptor alpha in breast cancer cells. Endocrinology, 147, 4831–4842 [[PubMed]
4. Ho S.M, et al. (2006). Estrogens and antiestrogens as etiological factors and therapeutics for prostate cancer. Ann. N. Y. Acad. Sci., 1089, 177–193 [[PubMed]
5. Horowitz M, et al. (1993). Effects of norethisterone on bone related biochemical variables and forearm bone mineral in post-menopausal osteoporosis. Clin. Endocrinol. (Oxf)., 39, 649–655 [[PubMed]
6. Jongen V, et al. (2009). Expression of estrogen receptor-alpha and -beta and progesterone receptor-A and -B in a large cohort of patients with endometrioid endometrial cancer. Gynecol. Oncol., 112, 537–542 [[PubMed]
7. Konstantinopoulos P.A, et al. (2003). Oestrogen receptor beta (ERbeta) is abundantly expressed in normal colonic mucosa, but declines in colon adenocarcinoma paralleling the tumour’s dedifferentiation. Eur. J. Cancer, 39, 1251–1258 [[PubMed]
8. Kuiper G.G, et al. (1997). Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors alpha and beta. Endocrinology, 138, 863–870 [[PubMed]
9. Liu M.M, et al. (2002). Opposing action of estrogen receptors alpha and beta on cyclin D1 gene expression. J. Biol. Chem., 277, 24353–24360 [[PubMed]
10. Krieg M, et al. (1993). Effect of aging on endogenous level of 5 alpha-dihydrotestosterone, testosterone, estradiol, and estrone in epithelium and stroma of normal and hyperplastic human prostate. J. Clin. Endocrinol. Metab., 77, 375–381 [[PubMed]
11. Harris R, et al. (2002). Screening for prostate cancer: an update of the evidence for the U.S. Preventive Services Task Force. Ann. Intern. Med., 137, 917–929 [[PubMed]
12. Rohrmann S, et al. (2007). Serum estrogen, but not testosterone, levels differ between black and white men in a nationally representative sample of Americans. J. Clin. Endocrinol. Metab., 92, 2519–2525 [[PubMed]
13. Briganti A. (2009) Oestrogens and prostate cancer: novel concepts about an old issue. Eur. Urol., 55, 543–545 [[PubMed][Google Scholar]
14. Leung Y.K, et al. (2011). Estrogen receptor β: switching to a new partner and escaping from estrogen. Sci. Signal., 4, pe19 [[PubMed]
15. Nakajima Y, et al. (2011). Estrogen regulates tumor growth through a nonclassical pathway that includes the transcription factors ERβ and KLF5. Sci. Signal., 4, ra22 [[PubMed]
16. Hu W.Y, et al. (2011). Estrogen-initiated transformation of prostate epithelium derived from normal human prostate stem-progenitor cells. Endocrinology, 152, 2150–2163
17. Carruba G. (2007) Estrogen and prostate cancer: an eclipsed truth in an androgen-dominated scenario. J. Cell. Biochem., 102, 899–911 [[PubMed][Google Scholar]
18. Adams J.Y, et al. (2002). Expression of estrogen receptor beta in the fetal, neonatal, and prepubertal human prostate. Prostate, 52, 69–81 [[PubMed]
19. Lai J.S, et al. (2004). Metastases of prostate cancer express estrogen receptor-beta. Urology, 64, 814–820 [[PubMed]
20. Latil A, et al. (2001). Evaluation of androgen, estrogen (ER alpha and ER beta), and progesterone receptor expression in human prostate cancer by real-time quantitative reverse transcription-polymerase chain reaction assays. Cancer Res., 61, 1919–1926 [[PubMed]
21. Linja M.J, et al. (2003). Expression of ERalpha and ERbeta in prostate cancer. Prostate, 55, 180–186 [[PubMed]
22. Leav I, et al. (2001). Comparative studies of the estrogen receptors beta and alpha and the androgen receptor in normal human prostate glands, dysplasia, and in primary and metastatic carcinoma. Am. J. Pathol., 159, 79–92
23. Pasquali D, et al. (2001). Loss of estrogen receptor beta expression in malignant human prostate cells in primary cultures and in prostate cancer tissues. J. Clin. Endocrinol. Metab., 86, 2051–2055 [[PubMed]
24. Gallo D, et al. (2012). Estrogen receptor beta in cancer: an attractive target for therapy. Curr. Pharm. Des., 18, 2734–2757 [[PubMed]
25. McCarty K.S, Jr, et al. (1985). Estrogen receptor analyses. Correlation of biochemical and immunohistochemical methods using monoclonal antireceptor antibodies. Arch. Pathol. Lab. Med., 109, 716–721 [[PubMed]
26. Chandran U.R, et al. (2007). Gene expression profiles of prostate cancer reveal involvement of multiple molecular pathways in the metastatic process. BMC Cancer, 7, 64
27. Irizarry R.A, et al. (2003). Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics, 4, 249–264 [[PubMed]
28. Taylor B.S, et al. (2010). Integrative genomic profiling of human prostate cancer. Cancer Cell, 18, 11–22
29. Arnold J.T, et al. (2005). Comparative effects of DHEA vs. testosterone, dihydrotestosterone, and estradiol on proliferation and gene expression in human LNCaP prostate cancer cells. Am. J. Physiol. Endocrinol. Metab., 288, E573–E584 [[PubMed]
30. Wang Y, et al. (2000). Sex hormone-induced carcinogenesis in Rb-deficient prostate tissue. Cancer Res., 60, 6008–6017 [[PubMed]
31. Lord R.S, et al. (2002). Estrogen metabolism and the diet-cancer connection: rationale for assessing the ratio of urinary hydroxylated estrogen metabolites. Altern. Med. Rev., 7, 112–129 [[PubMed]
32. Bosland M.C. (2000) The role of steroid hormones in prostate carcinogenesis. J. Natl Cancer Inst. Monogr., 27, 39–66 [[PubMed][Google Scholar]
33. Hsing A.W, et al. (1993). Serological precursors of cancer: serum hormones and risk of subsequent prostate cancer. Cancer Epidemiol. Biomarkers Prev., 2, 27–32 [[PubMed]
34. Roddam A.W, et al. (2008). Endogenous sex hormones and prostate cancer: a collaborative analysis of 18 prospective studies. J. Natl Cancer Inst., 100, 170–183
35. Griffiths K. (2000) Estrogens and prostatic disease. International Prostate Health Council Study Group. Prostate, 45, 87–100 [[PubMed][Google Scholar]
36. Tsai C.J, et al. (2006). Sex steroid hormones in young manhood and the risk of subsequent prostate cancer: a longitudinal study in African-Americans and Caucasians (United States). Cancer Causes Control, 17, 1237–1244 [[PubMed]
37. Lau K.M, et al. (2000). Expression of estrogen receptor (ER)-alpha and ER-beta in normal and malignant prostatic epithelial cells: regulation by methylation and involvement in growth regulation. Cancer Res., 60, 3175–3182 [[PubMed]
38. Kim I.Y, et al. (2002). Raloxifene, a mixed estrogen agonist/antagonist, induces apoptosis in androgen-independent human prostate cancer cell lines. Cancer Res., 62, 5365–5369 [[PubMed]
39. Fixemer T, et al. (2003). Differential expression of the estrogen receptor beta (ERbeta) in human prostate tissue, premalignant changes, and in primary, metastatic, and recurrent prostatic adenocarcinoma. Prostate, 54, 79–87 [[PubMed]
40. Dondi D, et al. (2010). Estrogen receptor beta and the progression of prostate cancer: role of 5alpha-androstane-3beta,17beta-diol. Endocr. Relat. Cancer, 17, 731–742 [[PubMed]
41. Kim H.T, et al. (2002). Raloxifene, a mixed estrogen agonist/antagonist, induces apoptosis through cleavage of BAD in TSU-PR1 human cancer cells. J. Biol. Chem., 277, 32510–32515 [[PubMed]
42. Rossi V, et al. (2010). Raloxifene induces cell death and inhibits proliferation through multiple signaling pathways in prostate cancer cells expressing different levels of estrogen receptor alpha and beta. J. Cell Physiol., 226, 1334–1339 [[PubMed]
43. Mak P, et al. (2010). ERbeta impedes prostate cancer EMT by destabilizing HIF-1alpha and inhibiting VEGF-mediated snail nuclear localization: implications for Gleason grading. Cancer Cell, 17, 319–332
44. Sasaki M, et al. (2003). Hypermethylation can selectively silence multiple promoters of steroid receptors in cancers. Mol. Cell. Endocrinol., 202, 201–207 [[PubMed]
45. Celhay O, et al. (2010). Expression of estrogen related proteins in hormone refractory prostate cancer: association with tumor progression. J. Urol., 184, 2172–2178 [[PubMed]