Estrogens, alone or in combination with androgens, can induce aberrant growth and/or malignancy o... more Estrogens, alone or in combination with androgens, can induce aberrant growth and/or malignancy of the prostate gland. Squamous metaplasia is an abnormal form of prostatic epithelial differentiation elicited by exogenous estrogen alone. Estrogens elicit their effects via estrogen receptors (ER) in the prostate. Experiments using ERalpha and ERbeta null mice demonstrated that ERalpha, but not ERbeta is essential in the induction of prostatic squamous metaplasia. To determine the respective roles of epithelial versus stromal ERalpha in this response, the following tissue recombinants were constructed with prostatic epithelium (PRE) and stroma (S) from wild-type (wt) and ERalpha knockout (alphaERKO) mice: wt-S + wt-PRE, alphaERKO-S + alphaERKO-PRE, wt-S + alphaERKO-PRE and alphaERKO-S + wt-PRE. A metaplastic response to diethylstilbestrol (DES) was only observed in wt-S + wt-PRE tissue recombinants. Tissue recombinants containing alphaERKO-PRE and/or alphaERKO-S (alphaERKO-S + alphaERKO-PRE, wt-S + alphaERKO-PRE and alphaERKO-S + wt-PRE) failed to respond to DES. Therefore, full and uniform epithelial squamous metaplasia requires ERalpha in both the epithelium and stroma. Estradiol (E2) in combination with testosterone (T) was shown to be effective in inducing prostatic carcinogenesis in a tissue recombinant model composed of rat urogenital sinus mesenchyme plus mouse prostatic epithelium. A particularly efficient model of prostatic carcinogenesis in mice involves T + E2 treatment of mice bearing grafts of wild-type rat urogenital mesenchyme (rUGM) plus retinoblastoma gene (Rb) knockout (Rb-KO) prostatic epithelium (rUGM + Rb-KO-PRE). Such rUGM + Rb-KO-PRE tissue recombinants developed hyperplasia, atypical hyperplasia and invasive prostatic carcinoma with high efficiency. During carcinogenesis in rUGM + Rb-KO-PRE tissue recombinants, epithelial E-cadherin almost totally disappeared and epithelial PCNA labeling was elevated. These epithelial changes were associated with almost total loss of smooth muscle cells in the stroma. The results of this study demonstrate that the absence of the Rb tumor suppressor gene predisposes prostatic epithelial cells to hormonal carcinogenesis.
We previously showed that mesenchymal-epithelial interactions are necessary for the development o... more We previously showed that mesenchymal-epithelial interactions are necessary for the development of bladder smooth muscle. Specifically without bladder epithelium embryonic bladder mesenchyme does not differentiate into smooth muscle. We determine whether this process is specific to bladder epithelium or whether epithelial cells from other organ systems induce bladder mesenchyme to differentiate into smooth muscle, as well as whether epithelial age is an important variable. We recombined 14-day bladder mesenchyme before smooth muscle differentiation with rat epithelium from 14-day, 19-day, newborn and adult bladder, ureter, colon, ileum, stomach, cornea and epidermis. In addition, bladder epithelium was recombined with 14-day embryonic small intestinal, 14-day embryonic gastric and newborn seminal vesicle mesenchyme. All tissue recombinants were grafted under the renal capsule of an adult rat syngeneic host for 3 weeks. Immunohistochemical analysis with antibodies directed against smooth muscle alpha-actin revealed that all epithelial types studied induced bladder mesenchyme to differentiate into smooth muscle, although to different degrees. Induction of smooth muscle was independent of urothelial age. In addition, bladder epithelium induced intestinal, gastric and seminal vesicle mesenchyme to differentiate into smooth muscle and express an overall morphological pattern indicative of the bladder fibromuscular wall. The mechanism whereby urothelium induces bladder mesenchyme to differentiate into smooth muscle is not specific to embryonic urothelium. Older urothelium and heterotypic epithelium also induce smooth muscle differentiation. With the common use of bowel, stomach and ureteral segments for bladder augmentation it is important to understand the interaction of different types of epithelium with the native bladder.
ABSTRACT The clonally derived rat prostatic epithelial cell line NRP-152 was examined to determin... more ABSTRACT The clonally derived rat prostatic epithelial cell line NRP-152 was examined to determine its ability to differentiate in a tissue recombination model. NRP-152 cells alone, or combined with urogenital mesenchyme (UGM) or 10T1/2 fibroblasts, were grafted beneath the renal capsule of athymic rodent hosts. After 1 and 3 months, grafts were examined grossly and immunohistochemically. NRP-152 cells grafted alone formed small (10-25 mg) grafts without recognizable architecture. NRP-152 cells recombined with UGM formed larger grafts (50-100 mg after 28 days) containing glandular epithelium. Columnar luminal cells expressed cytokeratins 8 and 18 and rat prostatic secretory markers (DP-1 and DP-2). The epithelial ducts were surrounded by well-differentiated smooth muscle. The glandular epithelial cells were shown to be of rat origin. NRP-152 + 10T1/2 tissue recombinants formed small grafts (10-40 mg wet weight) after 1 month. The epithelial component of these grafts formed solid unbranched cords expressing cytokeratins 5 and 14; no glandular epithelial structures were observed. The stromal matrix was densely packed with a few cells expressing alpha-actin. A clonally derived prostatic epithelial cell line can form structurally and functionally normal prostatic tissue. This suggests that prostatic basal and luminal epithelial cells can be derived from a common progenitor.
Advances in Experimental Medicine and Biology, 1999
The urinary bladder has two major functions: 1) storage and 2) socially acceptable efficient empt... more The urinary bladder has two major functions: 1) storage and 2) socially acceptable efficient emptying.1 To achieve this end the bladder has evolved as a highly compliant muscular sac with a specialized epithelial lining able to accommodate extensive variations in internal dimensions while maintaining an impervious luminal surface. Mesenchymal-epithelial interactions play an important role in the differentiation of both the urothelium and the smooth muscle of the bladder.2 These interactions are also important in maintaining homeostasis and in responding to injury and insults in adulthood.3 There is evidence to support the idea that stromal-epithelial interactions also play important roles in both carcinoma of the urinary bladder4,5 and in fibrotic responses to urinary obstruction due to a number of causes.6 Such fibrotic responses lead to the formation of a non-compliant bladder, that is, an organ which is unable to expand sufficiently to maintain the low pressure internal environment required to both store urine and protect the kidneys from damage due to urinary reflux.
Prostatic carcinogenesis is a multistep process with well-documented stages. Although prostate ca... more Prostatic carcinogenesis is a multistep process with well-documented stages. Although prostate cancer is a major cause of mortality many small tumor foci never progress to form clinically significant disease, indicating that the disease process may be regulated at more than one level. Carcinogenesis is accompanied by increasing genetic damage to prostatic epithelial cells, however the pattern of genetic lesions is inconsistent. The differentiation of stromal cells surrounding tumors is more fibroblastic and less muscular than in normal prostate. The present communication reviews the roles of both genetic and, stromally derived, epigenetic effects on prostatic tumorigenesis.
1. Recent Prog Horm Res. 1983;39:559-98. Hormone-induced morphogenesis and growth: role of mesenc... more 1. Recent Prog Horm Res. 1983;39:559-98. Hormone-induced morphogenesis and growth: role of mesenchymal-epithelial interactions. Cunha GR, Chung LW, Shannon JM, Taguchi O, Fujii H. PMID: 6314450 [PubMed - indexed for MEDLINE]. Publication Types: ...
... currently in progress. Acknowledgementshe authors wish to thank Beth Reese, Mamie Sekkingstad... more ... currently in progress. Acknowledgementshe authors wish to thank Beth Reese, Mamie Sekkingstad, Caroline Damian, and Kathy McCor-mack for their technical assistance and Betty Aguilar for typing the manuscript. This study was ...
We previously showed that mesenchymal-epithelial interactions are necessary for the development o... more We previously showed that mesenchymal-epithelial interactions are necessary for the development of bladder smooth muscle. Specifically without bladder epithelium embryonic bladder mesenchyme does not differentiate into smooth muscle. We determine whether this process is specific to bladder epithelium or whether epithelial cells from other organ systems induce bladder mesenchyme to differentiate into smooth muscle, as well as whether epithelial age is an important variable. We recombined 14-day bladder mesenchyme before smooth muscle differentiation with rat epithelium from 14-day, 19-day, newborn and adult bladder, ureter, colon, ileum, stomach, cornea and epidermis. In addition, bladder epithelium was recombined with 14-day embryonic small intestinal, 14-day embryonic gastric and newborn seminal vesicle mesenchyme. All tissue recombinants were grafted under the renal capsule of an adult rat syngeneic host for 3 weeks. Immunohistochemical analysis with antibodies directed against smooth muscle alpha-actin revealed that all epithelial types studied induced bladder mesenchyme to differentiate into smooth muscle, although to different degrees. Induction of smooth muscle was independent of urothelial age. In addition, bladder epithelium induced intestinal, gastric and seminal vesicle mesenchyme to differentiate into smooth muscle and express an overall morphological pattern indicative of the bladder fibromuscular wall. The mechanism whereby urothelium induces bladder mesenchyme to differentiate into smooth muscle is not specific to embryonic urothelium. Older urothelium and heterotypic epithelium also induce smooth muscle differentiation. With the common use of bowel, stomach and ureteral segments for bladder augmentation it is important to understand the interaction of different types of epithelium with the native bladder.
Embryonic urogenital sinus mesenchyme (UGM) has been demonstrated previously to be a potent induc... more Embryonic urogenital sinus mesenchyme (UGM) has been demonstrated previously to be a potent inductor of prostatic morphogenesis and functional differentiation when associated with either embryonic urogenital sinus epithelium (UGE) or urothelium derived from adult urinary bladder (ABLE) and grown in male hosts. To determine the role of mesenchyme in prostatic acinar growth, homotypic tissue recombinants of 16‐day‐old embryonic mouse urogenital sinus mesenchyme and epithelium (UGMmouse + UGEmouse) were prepared in which the relative amounts of UGM to UGE were varied from approximately 0.1:1 to 100: 1. Recombinants were grown for 1 month in intact male hosts after which the amount of acinar growth was assessed by histological analysis and by determination of wet weight and DNA content. The latter criteria are valid measures of acinar content of histologically normal prostatic tissue because the rodent prostate is composed of greater than 80% ductalacinar tissue. From this analysis the amount of acinar growth was found to be determined by the amount of mesenchymal tissue and was independent of the amount of epithelium utilized to prepare the tissue recombinants. Similarly, the magnitude of growth in heterospecific rat‐mouse tissue recombinants prepared with urogenital sinus epithelium or adult bladder epithelium (ABLE) and urogenital sinus mesenchyme (UGMmouse + UGErat, UGMmouse + ABLErat UGMrat + UGEmouse, or UGMrat + ABLEmouse) is determined by the source of UGM. Although the initial DNA content per UGM is comparable between mouse and rat, the overall acinar growth induced by UGMrat is about 10‐fold greater than that induced by UGMmouse when recombined reciprocally with UGEmouse and UGErat, respectively. These results suggest that UGM is of fundamental importance as a regulator of prostatic epithelial growth. The relevance of this finding to the development of human benign prostatic hyperplasia is discussed.
ABSTRACT Hepatocyte nuclear factor-3alpha (HNF-3alpha), a member of the hepatocyte-forkhead-homol... more ABSTRACT Hepatocyte nuclear factor-3alpha (HNF-3alpha), a member of the hepatocyte-forkhead-homolog family of transcription factors, regulates gene expression in the endoderm-derived liver and lung. To determine if HNF-3alpha might also play a role in endodermal derivatives of the urogenital sinus, the expression of HNF-3alpha in male accessory sex organs was assessed by Northern blotting, in situ hybridization, and electrophoretic mobility shift analysis. RNA from the dorsolateral prostate (DP), ventral prostate (VP), anterior prostate (AP), seminal vesicle (SV), and bladder was compared with RNA from the liver and spleen as positive and negative controls, respectively. HNF-3alpha mRNA levels in the DP, VP, AP, and bladder were 20, 14, 5, and 6 times higher than the SV equivalent in the liver. HNF-3alpha mRNA was detected in 8 of 10 prostate epithelial cell lines (rat NRP 152 and 154, mouse Pr14, and human DU-145, PC3, LNCaP, ND-1, and BPH-1) but not in rat Dunning epithelial or mouse Pr12 cells. Addition of testosterone to castrated rats was found to prevent a drastic loss of HNF-3alpha mRNA in the VP. This result suggests that HNF-3alpha mRNA levels are at least indirectly regulated by testosterone. The HNF-3alpha mRNA is expressed in epithelial cells of the urogenital sinus derivatives VP, AP, DP, and bladder and Wolffian duct derivative, the SV. To confirm that functional HNF-3alpha protein is produced in the VP, electrophoretic mobility shift assays were performed with whole-cell extracts and high-affinity oligonucleotide (TTR-S) from the transthyretin promoter. Binding to TTR-S was disrupted when the extract was incubated with HNF-3alpha, but not with HNF-3beta, antibody. Taken together, the results using VP, AP, DP, SV, and bladder suggest that HNF-3alpha may play an important role in development and maintenance of urogenital tract epithelial cells.
Estrogens, alone or in combination with androgens, can induce aberrant growth and/or malignancy o... more Estrogens, alone or in combination with androgens, can induce aberrant growth and/or malignancy of the prostate gland. Squamous metaplasia is an abnormal form of prostatic epithelial differentiation elicited by exogenous estrogen alone. Estrogens elicit their effects via estrogen receptors (ER) in the prostate. Experiments using ERalpha and ERbeta null mice demonstrated that ERalpha, but not ERbeta is essential in the induction of prostatic squamous metaplasia. To determine the respective roles of epithelial versus stromal ERalpha in this response, the following tissue recombinants were constructed with prostatic epithelium (PRE) and stroma (S) from wild-type (wt) and ERalpha knockout (alphaERKO) mice: wt-S + wt-PRE, alphaERKO-S + alphaERKO-PRE, wt-S + alphaERKO-PRE and alphaERKO-S + wt-PRE. A metaplastic response to diethylstilbestrol (DES) was only observed in wt-S + wt-PRE tissue recombinants. Tissue recombinants containing alphaERKO-PRE and/or alphaERKO-S (alphaERKO-S + alphaERKO-PRE, wt-S + alphaERKO-PRE and alphaERKO-S + wt-PRE) failed to respond to DES. Therefore, full and uniform epithelial squamous metaplasia requires ERalpha in both the epithelium and stroma. Estradiol (E2) in combination with testosterone (T) was shown to be effective in inducing prostatic carcinogenesis in a tissue recombinant model composed of rat urogenital sinus mesenchyme plus mouse prostatic epithelium. A particularly efficient model of prostatic carcinogenesis in mice involves T + E2 treatment of mice bearing grafts of wild-type rat urogenital mesenchyme (rUGM) plus retinoblastoma gene (Rb) knockout (Rb-KO) prostatic epithelium (rUGM + Rb-KO-PRE). Such rUGM + Rb-KO-PRE tissue recombinants developed hyperplasia, atypical hyperplasia and invasive prostatic carcinoma with high efficiency. During carcinogenesis in rUGM + Rb-KO-PRE tissue recombinants, epithelial E-cadherin almost totally disappeared and epithelial PCNA labeling was elevated. These epithelial changes were associated with almost total loss of smooth muscle cells in the stroma. The results of this study demonstrate that the absence of the Rb tumor suppressor gene predisposes prostatic epithelial cells to hormonal carcinogenesis.
We previously showed that mesenchymal-epithelial interactions are necessary for the development o... more We previously showed that mesenchymal-epithelial interactions are necessary for the development of bladder smooth muscle. Specifically without bladder epithelium embryonic bladder mesenchyme does not differentiate into smooth muscle. We determine whether this process is specific to bladder epithelium or whether epithelial cells from other organ systems induce bladder mesenchyme to differentiate into smooth muscle, as well as whether epithelial age is an important variable. We recombined 14-day bladder mesenchyme before smooth muscle differentiation with rat epithelium from 14-day, 19-day, newborn and adult bladder, ureter, colon, ileum, stomach, cornea and epidermis. In addition, bladder epithelium was recombined with 14-day embryonic small intestinal, 14-day embryonic gastric and newborn seminal vesicle mesenchyme. All tissue recombinants were grafted under the renal capsule of an adult rat syngeneic host for 3 weeks. Immunohistochemical analysis with antibodies directed against smooth muscle alpha-actin revealed that all epithelial types studied induced bladder mesenchyme to differentiate into smooth muscle, although to different degrees. Induction of smooth muscle was independent of urothelial age. In addition, bladder epithelium induced intestinal, gastric and seminal vesicle mesenchyme to differentiate into smooth muscle and express an overall morphological pattern indicative of the bladder fibromuscular wall. The mechanism whereby urothelium induces bladder mesenchyme to differentiate into smooth muscle is not specific to embryonic urothelium. Older urothelium and heterotypic epithelium also induce smooth muscle differentiation. With the common use of bowel, stomach and ureteral segments for bladder augmentation it is important to understand the interaction of different types of epithelium with the native bladder.
ABSTRACT The clonally derived rat prostatic epithelial cell line NRP-152 was examined to determin... more ABSTRACT The clonally derived rat prostatic epithelial cell line NRP-152 was examined to determine its ability to differentiate in a tissue recombination model. NRP-152 cells alone, or combined with urogenital mesenchyme (UGM) or 10T1/2 fibroblasts, were grafted beneath the renal capsule of athymic rodent hosts. After 1 and 3 months, grafts were examined grossly and immunohistochemically. NRP-152 cells grafted alone formed small (10-25 mg) grafts without recognizable architecture. NRP-152 cells recombined with UGM formed larger grafts (50-100 mg after 28 days) containing glandular epithelium. Columnar luminal cells expressed cytokeratins 8 and 18 and rat prostatic secretory markers (DP-1 and DP-2). The epithelial ducts were surrounded by well-differentiated smooth muscle. The glandular epithelial cells were shown to be of rat origin. NRP-152 + 10T1/2 tissue recombinants formed small grafts (10-40 mg wet weight) after 1 month. The epithelial component of these grafts formed solid unbranched cords expressing cytokeratins 5 and 14; no glandular epithelial structures were observed. The stromal matrix was densely packed with a few cells expressing alpha-actin. A clonally derived prostatic epithelial cell line can form structurally and functionally normal prostatic tissue. This suggests that prostatic basal and luminal epithelial cells can be derived from a common progenitor.
Advances in Experimental Medicine and Biology, 1999
The urinary bladder has two major functions: 1) storage and 2) socially acceptable efficient empt... more The urinary bladder has two major functions: 1) storage and 2) socially acceptable efficient emptying.1 To achieve this end the bladder has evolved as a highly compliant muscular sac with a specialized epithelial lining able to accommodate extensive variations in internal dimensions while maintaining an impervious luminal surface. Mesenchymal-epithelial interactions play an important role in the differentiation of both the urothelium and the smooth muscle of the bladder.2 These interactions are also important in maintaining homeostasis and in responding to injury and insults in adulthood.3 There is evidence to support the idea that stromal-epithelial interactions also play important roles in both carcinoma of the urinary bladder4,5 and in fibrotic responses to urinary obstruction due to a number of causes.6 Such fibrotic responses lead to the formation of a non-compliant bladder, that is, an organ which is unable to expand sufficiently to maintain the low pressure internal environment required to both store urine and protect the kidneys from damage due to urinary reflux.
Prostatic carcinogenesis is a multistep process with well-documented stages. Although prostate ca... more Prostatic carcinogenesis is a multistep process with well-documented stages. Although prostate cancer is a major cause of mortality many small tumor foci never progress to form clinically significant disease, indicating that the disease process may be regulated at more than one level. Carcinogenesis is accompanied by increasing genetic damage to prostatic epithelial cells, however the pattern of genetic lesions is inconsistent. The differentiation of stromal cells surrounding tumors is more fibroblastic and less muscular than in normal prostate. The present communication reviews the roles of both genetic and, stromally derived, epigenetic effects on prostatic tumorigenesis.
1. Recent Prog Horm Res. 1983;39:559-98. Hormone-induced morphogenesis and growth: role of mesenc... more 1. Recent Prog Horm Res. 1983;39:559-98. Hormone-induced morphogenesis and growth: role of mesenchymal-epithelial interactions. Cunha GR, Chung LW, Shannon JM, Taguchi O, Fujii H. PMID: 6314450 [PubMed - indexed for MEDLINE]. Publication Types: ...
... currently in progress. Acknowledgementshe authors wish to thank Beth Reese, Mamie Sekkingstad... more ... currently in progress. Acknowledgementshe authors wish to thank Beth Reese, Mamie Sekkingstad, Caroline Damian, and Kathy McCor-mack for their technical assistance and Betty Aguilar for typing the manuscript. This study was ...
We previously showed that mesenchymal-epithelial interactions are necessary for the development o... more We previously showed that mesenchymal-epithelial interactions are necessary for the development of bladder smooth muscle. Specifically without bladder epithelium embryonic bladder mesenchyme does not differentiate into smooth muscle. We determine whether this process is specific to bladder epithelium or whether epithelial cells from other organ systems induce bladder mesenchyme to differentiate into smooth muscle, as well as whether epithelial age is an important variable. We recombined 14-day bladder mesenchyme before smooth muscle differentiation with rat epithelium from 14-day, 19-day, newborn and adult bladder, ureter, colon, ileum, stomach, cornea and epidermis. In addition, bladder epithelium was recombined with 14-day embryonic small intestinal, 14-day embryonic gastric and newborn seminal vesicle mesenchyme. All tissue recombinants were grafted under the renal capsule of an adult rat syngeneic host for 3 weeks. Immunohistochemical analysis with antibodies directed against smooth muscle alpha-actin revealed that all epithelial types studied induced bladder mesenchyme to differentiate into smooth muscle, although to different degrees. Induction of smooth muscle was independent of urothelial age. In addition, bladder epithelium induced intestinal, gastric and seminal vesicle mesenchyme to differentiate into smooth muscle and express an overall morphological pattern indicative of the bladder fibromuscular wall. The mechanism whereby urothelium induces bladder mesenchyme to differentiate into smooth muscle is not specific to embryonic urothelium. Older urothelium and heterotypic epithelium also induce smooth muscle differentiation. With the common use of bowel, stomach and ureteral segments for bladder augmentation it is important to understand the interaction of different types of epithelium with the native bladder.
Embryonic urogenital sinus mesenchyme (UGM) has been demonstrated previously to be a potent induc... more Embryonic urogenital sinus mesenchyme (UGM) has been demonstrated previously to be a potent inductor of prostatic morphogenesis and functional differentiation when associated with either embryonic urogenital sinus epithelium (UGE) or urothelium derived from adult urinary bladder (ABLE) and grown in male hosts. To determine the role of mesenchyme in prostatic acinar growth, homotypic tissue recombinants of 16‐day‐old embryonic mouse urogenital sinus mesenchyme and epithelium (UGMmouse + UGEmouse) were prepared in which the relative amounts of UGM to UGE were varied from approximately 0.1:1 to 100: 1. Recombinants were grown for 1 month in intact male hosts after which the amount of acinar growth was assessed by histological analysis and by determination of wet weight and DNA content. The latter criteria are valid measures of acinar content of histologically normal prostatic tissue because the rodent prostate is composed of greater than 80% ductalacinar tissue. From this analysis the amount of acinar growth was found to be determined by the amount of mesenchymal tissue and was independent of the amount of epithelium utilized to prepare the tissue recombinants. Similarly, the magnitude of growth in heterospecific rat‐mouse tissue recombinants prepared with urogenital sinus epithelium or adult bladder epithelium (ABLE) and urogenital sinus mesenchyme (UGMmouse + UGErat, UGMmouse + ABLErat UGMrat + UGEmouse, or UGMrat + ABLEmouse) is determined by the source of UGM. Although the initial DNA content per UGM is comparable between mouse and rat, the overall acinar growth induced by UGMrat is about 10‐fold greater than that induced by UGMmouse when recombined reciprocally with UGEmouse and UGErat, respectively. These results suggest that UGM is of fundamental importance as a regulator of prostatic epithelial growth. The relevance of this finding to the development of human benign prostatic hyperplasia is discussed.
ABSTRACT Hepatocyte nuclear factor-3alpha (HNF-3alpha), a member of the hepatocyte-forkhead-homol... more ABSTRACT Hepatocyte nuclear factor-3alpha (HNF-3alpha), a member of the hepatocyte-forkhead-homolog family of transcription factors, regulates gene expression in the endoderm-derived liver and lung. To determine if HNF-3alpha might also play a role in endodermal derivatives of the urogenital sinus, the expression of HNF-3alpha in male accessory sex organs was assessed by Northern blotting, in situ hybridization, and electrophoretic mobility shift analysis. RNA from the dorsolateral prostate (DP), ventral prostate (VP), anterior prostate (AP), seminal vesicle (SV), and bladder was compared with RNA from the liver and spleen as positive and negative controls, respectively. HNF-3alpha mRNA levels in the DP, VP, AP, and bladder were 20, 14, 5, and 6 times higher than the SV equivalent in the liver. HNF-3alpha mRNA was detected in 8 of 10 prostate epithelial cell lines (rat NRP 152 and 154, mouse Pr14, and human DU-145, PC3, LNCaP, ND-1, and BPH-1) but not in rat Dunning epithelial or mouse Pr12 cells. Addition of testosterone to castrated rats was found to prevent a drastic loss of HNF-3alpha mRNA in the VP. This result suggests that HNF-3alpha mRNA levels are at least indirectly regulated by testosterone. The HNF-3alpha mRNA is expressed in epithelial cells of the urogenital sinus derivatives VP, AP, DP, and bladder and Wolffian duct derivative, the SV. To confirm that functional HNF-3alpha protein is produced in the VP, electrophoretic mobility shift assays were performed with whole-cell extracts and high-affinity oligonucleotide (TTR-S) from the transthyretin promoter. Binding to TTR-S was disrupted when the extract was incubated with HNF-3alpha, but not with HNF-3beta, antibody. Taken together, the results using VP, AP, DP, SV, and bladder suggest that HNF-3alpha may play an important role in development and maintenance of urogenital tract epithelial cells.
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