Volume 24, Issue 8 pp. 632-638

Original Article: Laboratory Investigation

Expression profile of urothelial transcription factors in bladder biopsies with interstitial cystitis

Kanya Kaga,

Kanya Kaga

Department of Urology, Continence Center, Dokkyo Medical University, Mibu, Tochigi, Japan

Department of Urology, Graduate School of Medicine, Chiba University, Chiba, Japan

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Ken-ichi Inoue,

Corresponding Author

Ken-ichi Inoue

[email protected]

Center for Research Support, Dokkyo Medical University, Mibu, Tochigi, Japan

Correspondence: Ken-ichi Inoue Ph.D., Center for Research Support, Dokkyo Medical University, 880 Kitakobayashi, Mibu, Tochigi 321-0293, Japan. Email: [email protected]Search for more papers by this author

Mayuko Kaga,

Mayuko Kaga

Department of Urology, Continence Center, Dokkyo Medical University, Mibu, Tochigi, Japan

Department of Urology, Graduate School of Medicine, Chiba University, Chiba, Japan

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Tomohiko Ichikawa,

Tomohiko Ichikawa

Department of Urology, Graduate School of Medicine, Chiba University, Chiba, Japan

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Tomonori Yamanishi,

Tomonori Yamanishi

Department of Urology, Continence Center, Dokkyo Medical University, Mibu, Tochigi, Japan

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Kanya Kaga,

Kanya Kaga

Department of Urology, Continence Center, Dokkyo Medical University, Mibu, Tochigi, Japan

Department of Urology, Graduate School of Medicine, Chiba University, Chiba, Japan

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Ken-ichi Inoue,

Corresponding Author

Ken-ichi Inoue

[email protected]

Center for Research Support, Dokkyo Medical University, Mibu, Tochigi, Japan

Mayuko Kaga,

Mayuko Kaga

Department of Urology, Continence Center, Dokkyo Medical University, Mibu, Tochigi, Japan

Department of Urology, Graduate School of Medicine, Chiba University, Chiba, Japan

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Tomohiko Ichikawa,

Tomohiko Ichikawa

Department of Urology, Graduate School of Medicine, Chiba University, Chiba, Japan

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Tomonori Yamanishi,

Tomonori Yamanishi

Department of Urology, Continence Center, Dokkyo Medical University, Mibu, Tochigi, Japan

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First published: 18 June 2017

https://doi.org/10.1111/iju.13391

Citations: 6

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Abstract

Objectives

To characterize interstitial cystitis pathology based on the expression profile of urothelial tissue-specific master transcription factors.

Methods

Bladder carcinoma cell lines derived from the urothelial stem cells (epithelial or mesenchymal) were used to identify candidate urothelial master transcription factors. Gene expression was measured with quantitative reverse transcription polymerase chain reaction. From the initial screening of 170 transcription factors (human homologs of Drosophila segmentation genes and known master transcription factors from a database), 28 transcription factors were selected. Subsequently, messenger ribonucleic acid from bladder biopsies of interstitial cystitis patients was purified, and gene expression levels of known urothelial marker genes and candidate master transcription factors were measured. Multivariate expression data were analyzed with spss software.

Results

Factor analysis decomposed the expression profile into four axes: principal axis 1 included retinoic acid receptors and 17 candidate master transcription factors. Principal axis 2 included KRT5 and five candidates. Principal axis 3 included transcription factor TP63 and two candidates. Principal axis 4 included SHH and two candidates. Principal component analysis segregated biopsies from Hunner's lesion in the principal component 1 (retinoic acid)/principal component 2 (SOX13)/principal component 3 (TP63) space.

Conclusions

Urothelial master transcription factors could serve as novel diagnostic markers and potentially explain the molecular pathology of interstitial cystitis.

Supporting Information

Filename	Description
iju13391-sup-0001-FigureS1.docxWord document, 320.6 KB	Figure S1. (a) Six bladder carcinoma cell lines were used for the screening and whole-cell protein lysates were extracted. Each well was loaded with 50 μg protein, and western blotting was carried out for TP63, SHH, E-cadherin and GAPDH. Note that the three cell lines, 5637, HuB6 and Hub15, synthesized abundant E-cadherin as well as TP63, a master regulator of stratified epithelium. “Epi” denotes classification into E-type. GAPDH serves as an internal control. (b) Strategy for master TF screening. We chose 179 TFs for initial screening, which include developmental regulators evolutionary conserved from Drosophila melanogaster and known master TFs (bladder, human mammary epithelial cells and colon crypt biopsies. Master TFs are under feed-forward regulation of their own super enhancers, and maintain exceptionally high expression of their coding genes. Therefore, we used mRNA expression level as a screening criteria.
iju13391-sup-0002-FigureS2.xlsxMS Excel, 35.8 KB	Figure S2. Urothelial master TF candidates identified from initial screening. Bladder carcinoma cell lines are classified either as E (epithelial) or as M (mesenchymal) type.
iju13391-sup-0003-TableS1.xlsxMS Excel, 54.7 KB	Table S1. Sequences of oligos and fluorescent probes used in the present study.
iju13391-sup-0004-TableS2.xlsxMS Excel, 21.6 KB	Table S2. Taqman probes used in the present study.
iju13391-sup-0005-TableS3.xlsxMS Excel, 41.9 KB	Table S3. List of clinical parameters as well as gene expressions for urothelial master transcription factor candidates in interstitial cystitis biopsies.
iju13391-sup-0006-TableS4.xlsxMS Excel, 68.7 KB	Table S4. List of clinical parameters as well as gene expressions for known urothelial markers in interstitial cystitis biopsies.
iju13391-sup-0007-TableS5.xlsxMS Excel, 41.8 KB	Table S5. Pearson's correlations among distinct mRNA expressions in interstitial cystitis biopsies.
iju13391-sup-0008-AppendixS1.docxWord document, 93.6 KB	Appendix S1. SPSS output file for principal axis factoring.
iju13391-sup-0009-AppendixS2.docxWord document, 38.6 KB	Appendix S2. SPSS output analysis for principal component analysis.

Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

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Citing Literature

Volume24, Issue8

August 2017

Pages 632-638

Expression profile of urothelial transcription factors in bladder biopsies with interstitial cystitis

Abstract

Objectives

Methods

Results

Conclusions

Supporting Information

References

Citing Literature

References

Information

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Expression profile of urothelial transcription factors in bladder biopsies with interstitial cystitis

Abstract

Objectives

Methods

Results

Conclusions

Supporting Information

References

Citing Literature

References

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