Skip to main content

Advertisement

SpringerLink
Log in

Epithelial-to-mesenchymal transition based diagnostic and prognostic signature markers in non-muscle invasive and muscle invasive bladder cancer patients

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Background

Diagnostic and prognostic significance of epithelial-to-mesenchymal transition (EMT) associated biomarkers are evaluated in a cohort of NMIBC (non-muscle invasive bladder cancer) and MIBC (muscle invasive bladder cancer) patients.

Methods and results

Real-time quantitative polymerase chain reaction (RT-qPCR) and immunohistochemical (IHC) staining were carried out in 100 tumor specimens (59 NMIBC and 41 MIBC). The expressions of the epithelial marker, mesenchymal markers and EMT-activating transcription factors (EMT-ATFs) were determined at transcriptome and protein level followed by their statistical associations with clinicohistopathological variables of the patients. Transcriptomic expression analysis showed statistical relevance of tumor stage with increased Twist and Zeb-1; tumor type with reduced E-cadherin and increased Snail; and smoking/tobacco chewing status (S/TC) of patients with increased N-cadherin and Snail in NMIBC patients. Tumor grade with reduced message E-cadherin, gain of N-cadherin, Snail, Twist and Zeb-1; patients’ age with reduced E-cadherin and Twist gain; and tumor type with increased message N-cadherin exhibited associations in MIBC patients. Protein expression analysis identified statistical relevance of tumor grade with nuclear gain of Snail and Twist; and nuclear gain of Slug with S/TC status of NMIBC patients. Novel gain of membranous Vimentin deduced association with patients’ age in MIBC patients. Survival analysis identified novel Vimentin as the positive predictor of short progression free survival (PFS) and short overall survival (OS) in MIBC patients. Study established altered EMT profile as the independent negative predictor of short recurrence free survival (RFS) in NMIBC patients and positive predictor of short PFS and OS in MIBC patients.

Conclusions

EMT associated biomarkers could provide diagnostic and prognostic risk stratification and hence could be of importance in the clinical management of bladder cancer patients.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Saginala K, Barsouk A, Aluru JS, Rawla P, Padala SA, Barsouk A (2020) Epidemiology of bladder cancer. Med Sci (Basel) 8(1):15

    CAS  Google Scholar 

  2. Garg M (2014) Prognostic and therapeutic applications of the molecular events in clinical management of urothelial carcinoma of bladder. J Exp Ther Oncol 10(4):301–316

    CAS  PubMed  Google Scholar 

  3. Garg M (2015) Urothelial cancer stem cells and epithelial plasticity: current concepts and therapeutic implications in bladder cancer. Cancer Metastasis Rev 34(4):691–770. https://doi.org/10.1007/s10555-015-9589-6

    Article  CAS  PubMed  Google Scholar 

  4. Siegel RL, Miller KD, Jemal A (2020) Cancer statistics, 2020. CA Cancer J Clin 70(1):7–30. https://doi.org/10.3322/caac.21590

    Article  PubMed  Google Scholar 

  5. McConkey DJ, Choi W, Marquis L, Martin F, Williams MB, Shah J, Svatek R, Das A, Adam L, Kamat A, Radtke AS, Dinney C (2009) Role of epithelial-to-mesenchymal transition (EMT) in drug sensitivity and metastasis in bladder cancer. Cancer Metastasis Rev 28:335–344. https://doi.org/10.1007/s10555-009-9194-7

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Ribatti D, Tamma R, Annese T (2020) Epithelial-mesenchymal transition in cancer: a historical overview. Transl Oncol 13(6):100773. https://doi.org/10.1016/j.tranon.2020.100773

    Article  PubMed  PubMed Central  Google Scholar 

  7. Xiong Y, Zu X, Wang L et al (2021) The VIM-AS1/miR-655/ZEB1 axis modulates bladder cancer cell metastasis by regulating epithelial–mesenchymal transition. Cancer Cell Int 21:233. https://doi.org/10.1186/s12935-021-01841-y

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Thiery JP, Acloque H, Ruby Huang YJ, Nieto MA (2009) Epithelial-mesenchymal transitions in development and disease. Cell 139(5):871–890. https://doi.org/10.1016/j.cell.2009.11.007

    Article  CAS  PubMed  Google Scholar 

  9. Garg M (2018) Epithelial plasticity and metastatic Cascade. Expert Opin Ther Targets 22(1):5–7. https://doi.org/10.1080/14728222.2018.1407312

    Article  PubMed  Google Scholar 

  10. Wu K, Zeng J, Zhou J et al (2013) Slug contributes to cadherin switch and malignant progression in muscle-invasive bladder cancer development. Urol Oncol 31(8):1751–1760. https://doi.org/10.1016/j.urolonc.2012.02.001

    Article  CAS  PubMed  Google Scholar 

  11. Loh CY, Chai JY, Tang TF et al (2019) The E-cadherin and N-cadherin switch in epithelial-to-mesenchymal transition: signaling, therapeutic implications, and challenges. Cells 8(10):1118. https://doi.org/10.3390/cells8101118

    Article  CAS  PubMed Central  Google Scholar 

  12. Soares IG, Chartoumpekis DV, Kyriazopoulou V, Zaravinos A (2020) EMT factors and metabolic pathways in cancer. Front Oncol. https://doi.org/10.3389/fonc.2020.00499

    Article  PubMed  PubMed Central  Google Scholar 

  13. Usman S, Waseem NH, Nguyen TKN et al (2021) Vimentin is at the heart of epithelial mesenchymal transition (EMT) mediated metastasis. Cancers (Basel) 13(19):4985. https://doi.org/10.3390/cancers13194985

    Article  CAS  Google Scholar 

  14. Garg M (2013) Epithelial-mesenchymal transition—activating transcription factors—multifunctional regulators in cancer. World J Stem Cells 5(4):188–195. https://doi.org/10.4252/wjsc.v5.i4.188

    Article  PubMed  PubMed Central  Google Scholar 

  15. Cheng L, Montironi R, Davidson D et al (2009) Staging and reporting of urothelial carcinoma of the urinary bladder. Mod Pathol 22:S70–S95. https://doi.org/10.1038/modpathol.2009.1

    Article  PubMed  Google Scholar 

  16. Singh R, Ansari JA, Maurya N, Mandhani A, Agrawal V, Garg M (2017) Epithelial-to-mesenchymal transition and its correlation with clinicopathological features in patients with urothelial carcinoma of the bladder. Clin Genitourin Cancer 15(2):e187–e197. https://doi.org/10.1016/j.clgc.2016.07.021

    Article  PubMed  Google Scholar 

  17. Kvikstad V, Mangrud OM, Gudlaugsson E et al (2019) Prognostic value and reproducibility of different microscopic characteristics in the WHO grading systems for pTa and pT1 urinary bladder urothelial carcinomas. Diagn Pathol 14:90. https://doi.org/10.1186/s13000-019-0868-3

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Xie Y, Li P, Gao Y et al (2017) Reduced E-cadherin expression is correlated with poor prognosis in patients with bladder cancer: a systematic review and meta-analysis. Oncotarget 8:62489–62499

    Article  Google Scholar 

  19. Baumgart E, Cohen MS, Silva Neto B et al (2007) Identification and prognostic significance of an epithelial-mesenchymal transition expression profile in human bladder tumors. Clin Cancer Res 13:1685–1694. https://doi.org/10.1158/1078-0432.CCR-06-2330

    Article  CAS  PubMed  Google Scholar 

  20. Zhang Y, Xu L, Li A, Han X (2019) The roles of ZEB1 in tumorigenic progression and epigenetic modifications. Biomed Pharmacother 110:400–408. https://doi.org/10.1016/j.biopha.2018.11.112

    Article  CAS  PubMed  Google Scholar 

  21. Favaretto RL, Bahadori A, Mathieu R et al (2017) Prognostic role of decreased E-cadherin expression in patients with upper tract urothelial carcinoma: a multi-institutional study. World J Urol 35(1):113–120. https://doi.org/10.1007/s00345-016-1835-1

    Article  CAS  PubMed  Google Scholar 

  22. Liang Z, Xie W, Wu R et al (2015) Inhibition of tobacco smoke-induced bladder MAPK activation and epithelial-mesenchymal transition in mice by curcumin. Int J Clin Exp Pathol 8(5):4503–4513 (26191140)

    PubMed  PubMed Central  Google Scholar 

  23. Bryan RT (2015) Cell adhesion and urothelial bladder cancer: the role of cadherin switching and related phenomena. Philos Trans R Soc Lond 370(1661):20140042. https://doi.org/10.1098/rstb.2014.0042

    Article  CAS  Google Scholar 

  24. Yu Q, Zhang K, Wang X, Liu X, Zhang Z (2010) Expression of transcription factors snail, slug and twist in human bladder carcinoma. J Exp Clin Cancer Res 29:119. https://doi.org/10.1186/1756-9966-29-119

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Singh R, Mandhani A, Agrawal V, Garg M (2019) Immunohistochemical studies to examine the diagnostic and prognostic implications of epithelial-to-mesenchymal transition in patients with urothelial carcinoma of the bladder. SN Comp Clin Med 1:277–286. https://doi.org/10.1007/s42399-018-0028-1

    Article  CAS  Google Scholar 

  26. Villarejo A, Cortés-Cabrera A, Molina-Ortíz P et al (2014) Differential role of snail1 and snail2 zinc fingers in E-cadherin repression and epithelial to mesenchymal transition. J Biol Chem 289(2):930–941. https://doi.org/10.1074/jbc.M113.528026

    Article  CAS  PubMed  Google Scholar 

  27. Hajra KM, Chen DY, Fearon ER (2002) The SLUG zinc-finger protein represses E-cadherin in breast cancer. Cancer Res 62:1613–1618

    CAS  PubMed  Google Scholar 

  28. lou V, Zaravinos A (2020) EMT factors and metabolic pathways in cancer. Front Oncol. https://doi.org/10.3389/fonc.2020.00499

    Article  PubMed  PubMed Central  Google Scholar 

  29. Fondrevelle ME, Kantelip B, Reiter RE et al (2009) The expression of twist has an impact on survival in human bladder cancer and is influenced by the smoking status. Urol Oncol 27:268–276. https://doi.org/10.1016/j.urolonc.2007.12.012

    Article  CAS  PubMed  Google Scholar 

  30. Bzerniak B, Dinney C, MacConkey D (2016) Origins of bladder cancer. Annu Rev Pathol 11:149–174. https://doi.org/10.1146/annurev-pathol-012513-104703

    Article  CAS  Google Scholar 

  31. Drápela S, Bouchal J, Jolly MK et al (2020) ZEB1: a critical regulator of cell plasticity, DNA damage response, and therapy resistance. Front Mol Biosci. https://doi.org/10.3389/fmolb.2020.00036

    Article  PubMed  PubMed Central  Google Scholar 

  32. Sánchez-Tilló E, Liu Y, de Barrios O et al (2012) EMT-activating transcription factors in cancer: beyond EMT and tumor invasiveness. Cell Mol Life Sci 69(20):3429–3456. https://doi.org/10.1007/s00018-012-1122-2

    Article  CAS  PubMed  Google Scholar 

  33. Garg M, Singh R (2019) Epithelial-to-mesenchymal transition: event and core associates in bladder cancer. Front Biosci Elite 11:150–165. https://doi.org/10.2741/E853

    Article  Google Scholar 

  34. Singh R, Mandhani A, Agrawal V, Garg M (2018) Positive correlation between matrix metalloproteinases and epithelial-to-mesenchymal transition and its association with clinical outcome in bladder cancer patients. Cancer Microenviron 11:23–39. https://doi.org/10.1007/s12307-017-0199-4

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

Authors thank Department of Science and Technology, Govt. of India for providing financial support to carry out research work (Research Grant No. SR/SO/HS-0113/2010). One of the authors, RS is thankful to University Grants Commission (UGC), Govt. of India for providing research fellowship.

Author information

Authors and Affiliations

  1. Department of Biochemistry, University of Lucknow, Lucknow, 226007, India

    R. Singh & M. Garg

  2. Department of Urology and Renal Transplantation, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, 226014, India

    U. P. Singh

  3. Department of Pathology, Sanjay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, 226014, India

    V. Agrawal

Authors
  1. R. Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. U. P. Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. V. Agrawal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Garg
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by RS and MG. The first draft of the manuscript was written by RS and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to M. Garg.

Ethics declarations

Conflict of interest

Authors disclose no potential conflict of interest.

Ethical approval

Studies were performed in accordance with the ethical standards laid down in the 1964 Declaration of Helsinki. Ethical clearance was obtained from Bioethics Cell, Institutional Ethics Committee (IEC) SGPGIMS, Lucknow, India (Reference No. 2014-166-CP-80).

Consent to participate

Analysis used anonymous data that were obtained after each patient agreed to participate in the study. A written informed consent was taken from each patient or immediate family member.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 24 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Singh, R., Singh, U.P., Agrawal, V. et al. Epithelial-to-mesenchymal transition based diagnostic and prognostic signature markers in non-muscle invasive and muscle invasive bladder cancer patients. Mol Biol Rep 49, 7541–7556 (2022). https://doi.org/10.1007/s11033-022-07563-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-022-07563-2

Keywords

Search

Navigation