Loss of the urothelial differentiation marker FOXA1 is associated with high grade, late stage bladder cancer and increased tumor proliferation

David Degraff, Peter E. Clark, Justin M. Cates, Hironobu Yamashita, Victoria L. Robinson, Xiuping Yu, Mark E. Smolkin, Sam S. Chang, Michael S. Cookson, Mary K. Herrick, Shahrokh F. Shariat, Gary D. Steinberg, Henry F. Frierson, Xue Ru Wu, Dan Theodorescu, Robert J. Matusik

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Abstract

Approximately 50% of patients with muscle-invasive bladder cancer (MIBC) develop metastatic disease, which is almost invariably lethal. However, our understanding of pathways that drive aggressive behavior of MIBC is incomplete. Members of the FOXA subfamily of transcription factors are implicated in normal urogenital development and urologic malignancies. FOXA proteins are implicated in normal urothelial differentiation, but their role in bladder cancer is unknown. We examined FOXA expression in commonly used in vitro models of bladder cancer and in human bladder cancer specimens, and used a novel in vivo tissue recombination system to determine the functional significance of FOXA1 expression in bladder cancer. Logistic regression analysis showed decreased FOXA1 expression is associated with increasing tumor stage (p<0.001), and loss of FOXA1 is associated with high histologic grade (p<0.001). Also, we found that bladder urothelium that has undergone keratinizing squamous metaplasia, a precursor to the development of squamous cell carcinoma (SCC) exhibited loss of FOXA1 expression. Furthermore, 81% of cases of SCC of the bladder were negative for FOXA1 staining compared to only 40% of urothelial cell carcinomas. In addition, we showed that a subpopulation of FOXA1 negative urothelial tumor cells are highly proliferative. Knockdown of FOXA1 in RT4 bladder cancer cells resulted in increased expression of UPK1B, UPK2, UPK3A, and UPK3B, decreased E-cadherin expression and significantly increased cell proliferation, while overexpression of FOXA1 in T24 cells increased E-cadherin expression and significantly decreased cell growth and invasion. In vivo recombination of bladder cancer cells engineered to exhibit reduced FOXA1 expression with embryonic rat bladder mesenchyme and subsequent renal capsule engraftment resulted in enhanced tumor proliferation. These findings provide the first evidence linking loss of FOXA1 expression with histological subtypes of MIBC and urothelial cell proliferation, and suggest an important role for FOXA1 in the malignant phenotype of MIBC.

Original languageEnglish (US)
Article numbere36669
JournalPloS one
Volume7
Issue number5
DOIs
StatePublished - May 10 2012

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Differentiation Antigens
Urinary Bladder Neoplasms
Muscle
Tumors
Cells
neoplasms
Cell proliferation
Cadherins
Neoplasms
bladder
Cell growth
Muscles
muscles
Urinary Bladder
Regression analysis
cadherins
Capsules
squamous cell carcinoma
Logistics
Rats

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Degraff, David ; Clark, Peter E. ; Cates, Justin M. ; Yamashita, Hironobu ; Robinson, Victoria L. ; Yu, Xiuping ; Smolkin, Mark E. ; Chang, Sam S. ; Cookson, Michael S. ; Herrick, Mary K. ; Shariat, Shahrokh F. ; Steinberg, Gary D. ; Frierson, Henry F. ; Wu, Xue Ru ; Theodorescu, Dan ; Matusik, Robert J. / Loss of the urothelial differentiation marker FOXA1 is associated with high grade, late stage bladder cancer and increased tumor proliferation. In: PloS one. 2012 ; Vol. 7, No. 5.
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abstract = "Approximately 50{\%} of patients with muscle-invasive bladder cancer (MIBC) develop metastatic disease, which is almost invariably lethal. However, our understanding of pathways that drive aggressive behavior of MIBC is incomplete. Members of the FOXA subfamily of transcription factors are implicated in normal urogenital development and urologic malignancies. FOXA proteins are implicated in normal urothelial differentiation, but their role in bladder cancer is unknown. We examined FOXA expression in commonly used in vitro models of bladder cancer and in human bladder cancer specimens, and used a novel in vivo tissue recombination system to determine the functional significance of FOXA1 expression in bladder cancer. Logistic regression analysis showed decreased FOXA1 expression is associated with increasing tumor stage (p<0.001), and loss of FOXA1 is associated with high histologic grade (p<0.001). Also, we found that bladder urothelium that has undergone keratinizing squamous metaplasia, a precursor to the development of squamous cell carcinoma (SCC) exhibited loss of FOXA1 expression. Furthermore, 81{\%} of cases of SCC of the bladder were negative for FOXA1 staining compared to only 40{\%} of urothelial cell carcinomas. In addition, we showed that a subpopulation of FOXA1 negative urothelial tumor cells are highly proliferative. Knockdown of FOXA1 in RT4 bladder cancer cells resulted in increased expression of UPK1B, UPK2, UPK3A, and UPK3B, decreased E-cadherin expression and significantly increased cell proliferation, while overexpression of FOXA1 in T24 cells increased E-cadherin expression and significantly decreased cell growth and invasion. In vivo recombination of bladder cancer cells engineered to exhibit reduced FOXA1 expression with embryonic rat bladder mesenchyme and subsequent renal capsule engraftment resulted in enhanced tumor proliferation. These findings provide the first evidence linking loss of FOXA1 expression with histological subtypes of MIBC and urothelial cell proliferation, and suggest an important role for FOXA1 in the malignant phenotype of MIBC.",
author = "David Degraff and Clark, {Peter E.} and Cates, {Justin M.} and Hironobu Yamashita and Robinson, {Victoria L.} and Xiuping Yu and Smolkin, {Mark E.} and Chang, {Sam S.} and Cookson, {Michael S.} and Herrick, {Mary K.} and Shariat, {Shahrokh F.} and Steinberg, {Gary D.} and Frierson, {Henry F.} and Wu, {Xue Ru} and Dan Theodorescu and Matusik, {Robert J.}",
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Degraff, D, Clark, PE, Cates, JM, Yamashita, H, Robinson, VL, Yu, X, Smolkin, ME, Chang, SS, Cookson, MS, Herrick, MK, Shariat, SF, Steinberg, GD, Frierson, HF, Wu, XR, Theodorescu, D & Matusik, RJ 2012, 'Loss of the urothelial differentiation marker FOXA1 is associated with high grade, late stage bladder cancer and increased tumor proliferation', PloS one, vol. 7, no. 5, e36669. https://doi.org/10.1371/journal.pone.0036669

Loss of the urothelial differentiation marker FOXA1 is associated with high grade, late stage bladder cancer and increased tumor proliferation. / Degraff, David; Clark, Peter E.; Cates, Justin M.; Yamashita, Hironobu; Robinson, Victoria L.; Yu, Xiuping; Smolkin, Mark E.; Chang, Sam S.; Cookson, Michael S.; Herrick, Mary K.; Shariat, Shahrokh F.; Steinberg, Gary D.; Frierson, Henry F.; Wu, Xue Ru; Theodorescu, Dan; Matusik, Robert J.

In: PloS one, Vol. 7, No. 5, e36669, 10.05.2012.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Loss of the urothelial differentiation marker FOXA1 is associated with high grade, late stage bladder cancer and increased tumor proliferation

AU - Degraff, David

AU - Clark, Peter E.

AU - Cates, Justin M.

AU - Yamashita, Hironobu

AU - Robinson, Victoria L.

AU - Yu, Xiuping

AU - Smolkin, Mark E.

AU - Chang, Sam S.

AU - Cookson, Michael S.

AU - Herrick, Mary K.

AU - Shariat, Shahrokh F.

AU - Steinberg, Gary D.

AU - Frierson, Henry F.

AU - Wu, Xue Ru

AU - Theodorescu, Dan

AU - Matusik, Robert J.

PY - 2012/5/10

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N2 - Approximately 50% of patients with muscle-invasive bladder cancer (MIBC) develop metastatic disease, which is almost invariably lethal. However, our understanding of pathways that drive aggressive behavior of MIBC is incomplete. Members of the FOXA subfamily of transcription factors are implicated in normal urogenital development and urologic malignancies. FOXA proteins are implicated in normal urothelial differentiation, but their role in bladder cancer is unknown. We examined FOXA expression in commonly used in vitro models of bladder cancer and in human bladder cancer specimens, and used a novel in vivo tissue recombination system to determine the functional significance of FOXA1 expression in bladder cancer. Logistic regression analysis showed decreased FOXA1 expression is associated with increasing tumor stage (p<0.001), and loss of FOXA1 is associated with high histologic grade (p<0.001). Also, we found that bladder urothelium that has undergone keratinizing squamous metaplasia, a precursor to the development of squamous cell carcinoma (SCC) exhibited loss of FOXA1 expression. Furthermore, 81% of cases of SCC of the bladder were negative for FOXA1 staining compared to only 40% of urothelial cell carcinomas. In addition, we showed that a subpopulation of FOXA1 negative urothelial tumor cells are highly proliferative. Knockdown of FOXA1 in RT4 bladder cancer cells resulted in increased expression of UPK1B, UPK2, UPK3A, and UPK3B, decreased E-cadherin expression and significantly increased cell proliferation, while overexpression of FOXA1 in T24 cells increased E-cadherin expression and significantly decreased cell growth and invasion. In vivo recombination of bladder cancer cells engineered to exhibit reduced FOXA1 expression with embryonic rat bladder mesenchyme and subsequent renal capsule engraftment resulted in enhanced tumor proliferation. These findings provide the first evidence linking loss of FOXA1 expression with histological subtypes of MIBC and urothelial cell proliferation, and suggest an important role for FOXA1 in the malignant phenotype of MIBC.

AB - Approximately 50% of patients with muscle-invasive bladder cancer (MIBC) develop metastatic disease, which is almost invariably lethal. However, our understanding of pathways that drive aggressive behavior of MIBC is incomplete. Members of the FOXA subfamily of transcription factors are implicated in normal urogenital development and urologic malignancies. FOXA proteins are implicated in normal urothelial differentiation, but their role in bladder cancer is unknown. We examined FOXA expression in commonly used in vitro models of bladder cancer and in human bladder cancer specimens, and used a novel in vivo tissue recombination system to determine the functional significance of FOXA1 expression in bladder cancer. Logistic regression analysis showed decreased FOXA1 expression is associated with increasing tumor stage (p<0.001), and loss of FOXA1 is associated with high histologic grade (p<0.001). Also, we found that bladder urothelium that has undergone keratinizing squamous metaplasia, a precursor to the development of squamous cell carcinoma (SCC) exhibited loss of FOXA1 expression. Furthermore, 81% of cases of SCC of the bladder were negative for FOXA1 staining compared to only 40% of urothelial cell carcinomas. In addition, we showed that a subpopulation of FOXA1 negative urothelial tumor cells are highly proliferative. Knockdown of FOXA1 in RT4 bladder cancer cells resulted in increased expression of UPK1B, UPK2, UPK3A, and UPK3B, decreased E-cadherin expression and significantly increased cell proliferation, while overexpression of FOXA1 in T24 cells increased E-cadherin expression and significantly decreased cell growth and invasion. In vivo recombination of bladder cancer cells engineered to exhibit reduced FOXA1 expression with embryonic rat bladder mesenchyme and subsequent renal capsule engraftment resulted in enhanced tumor proliferation. These findings provide the first evidence linking loss of FOXA1 expression with histological subtypes of MIBC and urothelial cell proliferation, and suggest an important role for FOXA1 in the malignant phenotype of MIBC.

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