Mouse 17 beta-hydroxysteroid dehydrogenase type 2 mRNA is predominantly expressed in hepatocytes and in surface epithelial cells of the gastrointestinal and urinary tracts

Mustonen, Mika; Poutanen, Matti; Kellokumpu, Sakari; Launoit, Yvan de; Isomaa, Veli; Vihko, R.; Vihko, Pirkko

doi:10.1677/jme.0.0200067

Cited by 48 publications

(23 citation statements)

References 40 publications

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“…Immunohistochemical data presented here indicate that 17β-HSD2 and 4 are localized predominantly within the luminal surface of the normal colonic epithelium, supporting previous reports of 17β-HSD2 expression in the gastrointestinal epithelium of the mouse (Mustonen et al, 1998), and human fetus (Takeyama et al, 2000). It would therefore appear that oestrogen inactivation by locally expressed 17β-HSD isozymes is a feature of normal gastrointestinal biology.…”

Section: Discussionsupporting

confidence: 89%

Oestrogen inactivation in the colon: analysis of the expression and regulation of 17 β -hydroxysteroid dehydrogenase isozymes in normal colon and colonic cancer

et al. 2000

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Summary Epidemiological data suggest that oestrogen contributes to the aetiology of colonic cancer. Furthermore, recent studies have suggested that local hormone metabolism may play a key role in determining colonic responsiveness to oestrogen. To further clarify this mechanism we have characterized the expression and regulation of isozymes of 17β-hydroxysteroid dehydrogenase (17β-HSD) in vitro and in situ. Immunohistochemistry was used to confirm expression of the type 2 and 4 isozymes of 17β-HSD (17β-HSD2 and 4) in normal colonic epithelial cells. Parallel studies suggested that both isozymes were abnormally expressed in colonic tumours and this was confirmed by Western blot analyses. Abnormal expression of 17β-HSD2 and 4 proteins was also observed in Caco-2, HT-29 and SW620 colonic cancer cell lines, although the overall pattern of oestrogen metabolism in these cells was similar to that seen in primary colonic mucosal tissue. The predominant activity (conversion of oestradiol to oestrone) was highest in Caco-2>SW620>HT-29, which correlated inversely with the rate of proliferation of the cell lines. Regulatory studies using SW620 cells indicated that the most potent stimulator of oestradiol to oestrone inactivation was the antiproliferative agent 1,25-dihydroxyvitamin D 3 (1,25D 3 ), whilst oestradiol itself inhibited 17β-HSD activity. Both oestradiol and 1,25D 3 decreased mRNA for 17β-HSD2 and 4. Data indicate that the high capacity for inactivation of oestrogens in the colon is associated with the presence of 17β-HSD2 and 4 in epithelial cells. MATERIALS AND METHODS Immunohistochemical studiesColonic tumour and paired normal mucosal tissue were obtained with agreement from the local ethical approval committee. Fivemicron thick, formalin-fixed tissue sections were cut and placed on coated glass slides. Sections were de-waxed and endogenous peroxidase activity quenched with 3% hydrogen peroxide. Sections were then incubated in donkey serum (Binding Site, Birmingham, UK) diluted 1/10 in PBS (15 min), followed by primary antibody diluted in PBS (1 hour). Antisera used were as follows: 17β-HSD2 monoclonal antiserum (1/500 dilution), a kind gift of Dr S Andersson (South Western Medical Center, Dallas, USA), was produced with a synthetic carboxyterminal peptide [C]RALRMP-NYKKKAT, corresponding to amino acids 375-387 in the human 17β-HSD2 protein; 17β-HSD4 monoclonal antibody (1:200 dilution), a kind gift of Dr J Adamski (GSF, Neuherberg, Germany) was prepared against the porcine 17β-HSD4 which cross-reacts with human, rat and mouse tissues. After washing, slides were incubated for 30 min with a biotinylated universal secondary antibody (Binding Site), diluted 1/100 in PBS, and binding visualised using ABC reagent (Binding Site) and 3,3′-diaminobenzidine (Sigma Chemical Co, Poole, UK). After staining, slides were washed and counterstained in Mayer's haematoxylin. Cell cultureColonic carcinoma cell lines (SW620, Caco-2 and HT-29) were routinely maintained in Dulbecco's Modified Eagles Medium (DMEM), supplemented wit...

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Section: Discussionsupporting

confidence: 89%

Oestrogen inactivation in the colon: analysis of the expression and regulation of 17 β -hydroxysteroid dehydrogenase isozymes in normal colon and colonic cancer

et al. 2000

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“…The present study revealed abundant expression of 17HSD type 2 mRNA in the surface epithelial cells of the normal colon and small intestinal tissues of both female and male subjects. Previous studies 16,27 have shown this enzyme to be present also in the small intestine of the human fetus and in rodents. The available data on substrate specificity for 17HSD type 2 shows that it efficiently inactivates E2, T and DHT into their corresponding inactive keto forms, 16 thus decreasing the estrogen effect in many tissues and organs.…”

Section: Discussionmentioning

confidence: 81%

“…It has been localized to the placenta, breast, endometrium and some epithelial cells of human and rodent origin not considered classical endocrine target tissues. [13][14][15][16] The presence of the mRNA for the enzyme in the small intestine, as revealed by Northern blot analysis and in situ hybridization, has been ascribed a protective role as it lowers the concentrations of active sex steroids. 17 Detailed understanding of the expression of the mRNA for this enzyme in the normal gastrointestinal tract, intestinal inflammatory disorders and neoplastic growth is therefore important.…”

Section: Abstract: Colon Carcinoma; Estrogen; Hydroxysteroid Dehydromentioning

confidence: 99%

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Downregulation of estrogen‐metabolizing 17β‐hydroxysteroid dehydrogenase Type 2 expression correlates inversely with Ki67 proliferation marker in colon‐cancer development

Oduwole

Isomaa

Nokelainen

et al. 2001

Intl Journal of Cancer

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The 17HSDs are a group of isozymes that catalyze the interconversion between high-activity 17␤-hydroxysteroids and low-activity 17-ketosteroids. In the present study, we characterized the expression of 17HSD types 1 and 2 in normal and malignant gastrointestinal tissues and cells. Using the colon as a model for cancer of the gastrointestinal tract, expression of the 17HSD enzymes in cancer development was studied and correlated with proliferation and differentiation markers as assessed by Ki67 and mucin staining, respectively. In normal colon and small intestine, 17HSD type 2 mRNA was expressed in the surface epithelial cells and, to a lesser extent, in the cryptal epithelial cells. In colon-cancer specimens, 17HSD type 2 expression was downregulated both in the tissues and in the cell lines and correlated inversely with the proliferation marker. No expression for the 17HSD type 1 enzyme was observed in normal or cancerous gastrointestinal tract tissues. In line with the expression studies, 17HSD activity measurements with colon cells showed that only the oxidative conversion of E2 to E1 was present, and Northern blot analysis showed the signal only for 17HSD type 2. Localization of the ERs ␣ and ␤, assessed by immunohistochemistry and in situ hybridization, showed the presence of ER␤ in the lamina propria of the colon. Our study shows that 17HSD type 2 expression is associated with the functional integrity of the gastrointestinal tract. The decrease in expression of the type 2 enzyme may increase estrogen influence in colon cancer. © 2002 Wiley-Liss, Inc. Key words: colon carcinoma; estrogen; hydroxysteroid dehydrogenase; cell proliferation; cell differentiationEpidemiologic studies 1,2 and in vitro studies with colon-cancer cell lines 3 suggest the involvement of estrogens in the development and progression of gastrointestinal cancer. [3][4][5] Contrary to this, however, is the indication that HRT is effective at reducing the incidence of colon cancer. 6 The mechanism underlying the sex steroid-mediated effects in normal gastrointestinal physiology and its involvement in neoplastic development and progression, if any, are far from clear.The 17HSDs play pivotal roles in the regulation of the physiologic activities of sex steroid hormones by catalyzing the oxidation or reduction of 17␤-hydroxy-and 17-ketosteroids, respectively. 7,8 17HSDs regulate the concentrations of the active sex steroids that bind to the respective steroid receptors to regulate gene expression in target cells. To date, at least 8 isoforms of the 17HSD enzyme with different substrate and cofactor specificities, tissue distributions and subcellular localizations have been characterized. 8,9 Of these isoforms, 17HSD type 1 and type 2 catalyze opposite reactions of estrogen metabolism. 17HSD type 1 is the predominant enzyme involved in estradiol biosynthesis (E1 to E2), and it is expressed in cells of placental and ovarian origin 10,11 as well as in breast tissue. 12 On the other hand, 17HSD type 2 catalyzes the oxidation of estrogens (E2 to E1...

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“…15 Preservation of RNA in the samples was confirmed by glyceraldehyde-3-phosphate-dehydrogenase (accession number [AC] X02231) antisense probe. 2 The epithelial and stromal signal densities of TMPRSS2 expression in the form of silver grains per area were measured from in situ hybridized sections with an MCID M4 3.0 digital image analyzer (Imaging Research, St. Catharines, ON, Canada), using a method similar to that described by Kainu et al 16 In prostate cancer samples, signal density was measured separately in benign and malignant epithelium and stroma from 5 separate, randomly picked, 40ϫ objective fields of specimens without knowledge of tumor grade or treatment.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

TheTMPRSS2 gene encoding transmembrane serine protease is overexpressed in a majority of prostate cancer patients: Detection of mutatedTMPRSS2 form in a case of aggressive disease

et al. 2001

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The serine protease TMPRSS2 gene expression was studied by in situ hybridization using benign prostatic hyperplasia and prostate cancer tissue samples from 32 patients. Expression of TMPRSS2 gene was higher in cancer cells than that in benign cells in 84% of the specimens containing both benign and malignant tissues. The TMPRSS2 mRNA level was significantly increased in poorly differentiated (p ‫؍‬ 0.014, n ‫؍‬ 7) and untreated (p ‫؍‬ 0.022, n ‫؍‬ 13) primary prostate adenocarcinomas compared to benign tissues. In addition, androgen-deprivation therapy significantly decreased the expression of TMPRSS2 in benign prostate tissue (p ‫؍‬ 0.07), which is in accordance with the androgen-inducible expression of the gene. The gene copy number of TMPRSS2, analyzed by competitively differential PCR, was duplicated in the malignant cells of about 38% of the prostate cancer patients analyzed. Thus, the increase in the gene copy number is probably not the primary reason for the detected overexpression of the TMPRSS2 gene. Mutations in the TMPRSS2 gene were screened using DNA isolated from paraffin-embedded prostate cancer tissues from 9 patients with aggressive prostate cancer and from 9 patients with nonaggressive disease. Thirteen exons covering the coding region were checked using enzymatic mutation detection and direct sequencing. One patient with aggressive prostate cancer carried a deletion and a stop codon in exon 11, leading to inactivation of the serine protease domain in TMPRSS2.

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Mouse 17 beta-hydroxysteroid dehydrogenase type 2 mRNA is predominantly expressed in hepatocytes and in surface epithelial cells of the gastrointestinal and urinary tracts

Cited by 48 publications

References 40 publications

Oestrogen inactivation in the colon: analysis of the expression and regulation of 17 β -hydroxysteroid dehydrogenase isozymes in normal colon and colonic cancer

Oestrogen inactivation in the colon: analysis of the expression and regulation of 17 β -hydroxysteroid dehydrogenase isozymes in normal colon and colonic cancer

Downregulation of estrogen‐metabolizing 17β‐hydroxysteroid dehydrogenase Type 2 expression correlates inversely with Ki67 proliferation marker in colon‐cancer development

TheTMPRSS2 gene encoding transmembrane serine protease is overexpressed in a majority of prostate cancer patients: Detection of mutatedTMPRSS2 form in a case of aggressive disease

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