Testosterone metabolism in primary cultures of epithelial cells and stroma from benign prostatic hyperplasia

Tsugaya, Masayuki; Habib, Fouad K.; Chisholm, G. D.; Ross, Margaret; Tozawa, Keiichi; Hayashi, Yutaro; Kohri, K; Tanaka, Shin

doi:10.1007/bf00304775

Cited by 18 publications

(22 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Human prostatic fibroblast and epithelial cells were cultured from prostate chips as previously described (Habib 1992, Tsugaya et al 1996. Verification of the purity of the fibroblast and epithelial cells was established by immunohistochemical staining employing a variety of primary monoclonal antibodies and by phase contrast microscopy as detailed in our earlier reports (Habib 1992, Tsugaya et al 1996.…”

Section: Primary Cultures Of Prostate Fibroblast and Epithelial Cellsmentioning

confidence: 99%

“…Verification of the purity of the fibroblast and epithelial cells was established by immunohistochemical staining employing a variety of primary monoclonal antibodies and by phase contrast microscopy as detailed in our earlier reports (Habib 1992, Tsugaya et al 1996.…”

Section: Primary Cultures Of Prostate Fibroblast and Epithelial Cellsmentioning

confidence: 99%

“…Subsequent steps, including extraction and separation of steroids, development of thin layer chromatography plates and the recovery of steroids, have all been described previously (Houston et al 1985a, Tsugaya et al 1996.…”

Section: Measurement Of 5 R Activitymentioning

confidence: 99%

“…The 5 -reductase assays were set up in tubes containing an NADPH generating system as described by Tsugaya et al (1996) and the reactions proceeded following the addition of 200 µl homogenate along with [ 3 H]testosterone solution (20 nM; 1 µCi). The tubes were incubated in a shaking water bath at 37 C for 30 min and the reactions were stopped by the addition of two volumes of diethylether containing 500 c.p.m.…”

Section: Measurement Of 5 R Activitymentioning

confidence: 99%

See 3 more Smart Citations

The localisation and expression of 5 alpha-reductase types I and II mRNAs in human hyperplastic prostate and in prostate primary cultures

Habib

Ross²,

Bayne³

et al. 1998

Journal of Endocrinology

View full text Add to dashboard Cite

The expression and localisation of mRNAs for 5 reductase Type I (5 R-I) and Type II (5 R-II) isoenzymes in human benign prostatic hyperplasia (BPH) were investigated by RT-PCR and by in situ hybridisation (ISH) using digoxigenin labelled riboprobes. In addition, we also examined the isoenzymes mRNA expression in primary BPH cultures of separated stroma/fibroblast and epithelial cells to determine whether primary cultures are appropriate models in which to investigate 5 R activity and regulation. The results demonstrated conclusively the presence of mRNA encoding both isoenzymes in all specimens so far examined. Additionally, the presence of a functional 5 R-I and -II activity in BPH was confirmed by enzyme assays. ISH studies localised the mRNA expression to both the fibroblast/stromal component as well as the epithelial cells of the hyperplastic tissue. In the glandular regions the expression for both isoenzymes was particularly strong in the basal layers of the epithelium whereas mRNA expression in the secretory cells was less pronounced. Expression of 5 R-I and -II mRNAs in fibroblast was on the other hand variable with high expression in some areas and little in others. These findings were supported by our primary culture experiments which demonstrated that both the fibroblast and epithelial cells maintain a capacity to express both isoenzymes in vitro. In the case of the fibroblast, the capacity to express the isoenzymes was maintained following the sequential passaging of the cells up to passage 6, after which the cells no longer expressed either isoenzyme.

show abstract

Section: Primary Cultures Of Prostate Fibroblast and Epithelial Cellsmentioning

confidence: 99%

Section: Primary Cultures Of Prostate Fibroblast and Epithelial Cellsmentioning

confidence: 99%

Section: Measurement Of 5 R Activitymentioning

confidence: 99%

Section: Measurement Of 5 R Activitymentioning

confidence: 99%

See 2 more Smart Citations

The localisation and expression of 5 alpha-reductase types I and II mRNAs in human hyperplastic prostate and in prostate primary cultures

Habib

Ross²,

Bayne³

et al. 1998

Journal of Endocrinology

View full text Add to dashboard Cite

show abstract

“…In addition, estrogens can be produced locally in the prostate gland via conversion of testosterone to 17␤-estradiol by aromatase expressed within the stroma (16). With an increased estrogen:androgen ratio, epithelial cells secrete TGF-␤, a pleiotropic factor that induces smooth muscle differentiation and increases the extracellular matrix (ECM) in the surrounding stromal cells (14,17).…”

mentioning

confidence: 99%

A role for epithelial-mesenchymal transition in the etiology of benign prostatic hyperplasia

Alonso-Magdalena

Brössner

Reiner

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

152

127

View full text Add to dashboard Cite

Benign prostatic hyperplasia (BPH) is usually described as a pathological proliferation of prostatic fibroblasts/myofibroblasts and epithelial cells. In the present study of BPH samples, we have made a morphological and immunohistochemical study of BPH prostatic sections using markers of proliferation, apoptosis, hormone receptors, and TGF-␤ signaling. We found no evidence of proliferation in the stroma but in the epithelium of some ducts; 0.7% of the basal and 0.4% of luminal cells were positive for Ki67 and PCNA. Androgen receptor and estrogen receptor beta (ER␤)1 and ER␤cx were abundant in both stromal and epithelial compartments but cells expressing ER␣ were very rare. What was very common in all BPH samples was the following: (i) regions of the ductal epithelium where the epithelial cells did not express E-cadherin, had lost their polarization, and become spindle shaped (the nuclei of these cells were strongly positive for pSmad 3 and Snail); and (ii) regions where the walls of the blood vessels were extremely thick and there was loss of endothelial layer. Loss of E-cadherin, increased pSmad 3, and high expression of Snail are all characteristic of epithelial-mesenchymal transition (EMT). We conclude that BPH is not a disease of prostatic stromal proliferation but rather of accumulation of mesenchymal-like cells derived from the prostatic epithelium and the endothelium. TGF-␤ is thought to play a key role in EMT. Our data suggests that TGF-␤/Smad should be considered as targets for treatment of BPH.proliferation ͉ TGF beta signaling

show abstract

Interaction of prostate epithelial cells from benign and malignant tumor tissue with bone-marrow stroma

et al. 1998

View full text Add to dashboard Cite

BACKGROUND Metastases of prostate cancer form selectively within the skeleton. To understand this metastatic spread, we studied the ability of prostate epithelial cells to grow and proliferate within the bone marrow, using primary coculture. METHODS Prostate epithelia and fibroblasts were prepared from men with benign prostatic hyperplasia (n = 13) and cancer of the prostate (n = 10). Confluent cultures of bone‐marrow stroma or fibroblast controls were prepared in 96‐well plates, and identical plates were treated with detergent to expose the extracellular matrix of the cells. Epithelial cells were seeded onto either cells or matrix, and their growth characteristics were determined by counting increases in colony size and number over time. Further experiments evaluated the effects on epithelial growth when cells were exposed to media conditioned by these stroma, using an MTT assay. RESULTS Results showed that for epithelial cells derived from malignant (or benign) tissue, the median value of the total area of colonies formed on bone‐marrow stroma was 2.1 (benign, 2.6) mm2, in contrast to 0.3 (benign, 0.4) mm2 or 0.25 (benign, 0) mm2 when these cells were cocultured with fibroblasts from benign or malignant prostates, respectively. Statistics indicated that growth was significantly greater on bone‐marrow stroma than on control stroma (P < 0.005). However, no significant stimulation of epithelial cell growth was seen when these epithelial cells were cultured on extracellular matrix from bone‐marrow stroma or when exposed to bone‐marrow stroma‐conditioned media in comparison to fibroblast controls. No statistical differences were found between the formation of colonies from malignant tissue in comparison to benign. CONCLUSIONS This system allows the investigation of bone‐marrow stroma colonization by primary prostate epithelial cells, and could be developed for the study of metastasis. Prostate 34:203–213, 1998. © 1998 Wiley‐Liss, Inc.

show abstract

Testosterone metabolism in primary cultures of epithelial cells and stroma from benign prostatic hyperplasia

Cited by 18 publications

References 32 publications

The localisation and expression of 5 alpha-reductase types I and II mRNAs in human hyperplastic prostate and in prostate primary cultures

The localisation and expression of 5 alpha-reductase types I and II mRNAs in human hyperplastic prostate and in prostate primary cultures

A role for epithelial-mesenchymal transition in the etiology of benign prostatic hyperplasia

Interaction of prostate epithelial cells from benign and malignant tumor tissue with bone-marrow stroma

Contact Info

Product

Resources

About