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Monitoring the response of urothelial precancerous lesions to Bacillus Calmette-Guerin at the proteome level in an in vivo rat model

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Abstract

Intravesical Bacillus Calmette-Guerin (BCG) is the best treatment modality for progression of non-muscle invasive bladder cancer. We aimed to monitor changes at the proteome level to identify putative protein biomarkers associated with the response of urothelial precancerous lesions to intravesical BCG treatment. The rats were divided into three groups (n = 10/group): control, non-treated, and BCG-treated groups. The non-treated and BCG-treated groups received N-methyl-N-nitrosourea intravesically. BCG Tice-strain was instilled into bladder in BCG-treated group. At the endpoint of experiment, all surviving rat bladders were collected and equally divided into two portions vertically from dome to neck. Half of each bladder was assessed immunohistopathologically and the other half was used for 2D-based comparative proteomic analysis. Differentially expressed proteins were validated by Western blot analysis. Precancerous lesions of bladder cancer were more common in non-treated group (77.8%) than in BCG-treated group (50%) and the control group (0%). Greater than twofold changes occurred in the expression of a number of proteins. Among them, Rab-GDIβ, aldehyde dehydrogenase 2 (ALDH2) and 14-3-3 zeta/delta were important since they were previously reported to be associated with cancer and their expression levels were found to be lower in BCG-treated group in comparison to the non-treated group. ALDH2 and 14-3-3 zeta/delta were also found to be highly expressed in the non-treated group compared to the control group. The down-regulation of these proteins and Rab-GDIβ was achieved with BCG; this result indicates that they may be used as putative biomarkers for monitoring changes in bladder carcinogenesis in response to BCG immunotherapy.

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Abbreviations

2D:

2-Dimensional

2-DE:

2D Gel electrophoresis

ALDH:

Aldehyde dehydrogenase

ALDH1:

Aldehyde dehydrogenase 1

ALDH2:

Aldehyde dehydrogenase 2

BC:

Bladder cancer

BCG:

Bacillus Calmette-Guerin

CIS:

Carcinoma in situ

G3PD:

Glyceraldehyde-3-phosphate dehydrogenase

IPG:

Immobilized pH gradient

LGIEN:

Low-grade intraepithelial neoplasia

MALDI:

Matrix assisted laser desorption ionization

MNU:

N-Methyl N-nitrosourea

NMIBC:

Non-muscle invasive bladder cancer

Rab-GDIβ:

Rab GDP dissociation inhibitor beta

SDS-PAGE:

SDS-Polyacrylamide gel electrophoresis

TOF:

Time of flight

TUR:

Transurethral resection

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Authors and Affiliations

  1. Department of Urology, Kocaeli University School of Medicine, Umuttepe Campus, 41380, Kocaeli, Turkey

    Kerem Teke, Ali Kemal Uslubas, Hasan Yilmaz & Ozdal Dillioglugil

  2. Department of Molecular Biology, Kocaeli University School of Medicine, Kocaeli, Turkey

    Nil Guzel, Murat Kasap & Gurler Akpinar

  3. Department of Pathology, Kocaeli University School of Medicine, Kocaeli, Turkey

    Demir Kursat Yildiz

Authors
  1. Kerem Teke
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  2. Nil Guzel
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  3. Ali Kemal Uslubas
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  4. Murat Kasap
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  5. Hasan Yilmaz
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  6. Gurler Akpinar
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  7. Demir Kursat Yildiz
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  8. Ozdal Dillioglugil
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Contributions

KT project development, data collection, data analysis, and manuscript writing/editing. NG data collection. AKU data collection. MK project development, data analysis, and manuscript writing/editing. HY project development, data analysis. GA project development, data analysis, and manuscript writing/editing. DKY data collection, manuscript writing. OD project development, data analysis, and manuscript writing/editing.

Corresponding author

Correspondence to Kerem Teke.

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Conflict of interest

The authors do not have any conflicts of interest to disclose.

Funding

This work was supported by the Scientific and Investigational Projects Reserve of the University of Kocaeli (Grant number: 2015/035).

Ethical approval

This study was approved by the Local Ethical Committee on Animal Research at Kocaeli University, approval number: 11/3-2015. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.

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Teke, K., Guzel, N., Uslubas, A.K. et al. Monitoring the response of urothelial precancerous lesions to Bacillus Calmette-Guerin at the proteome level in an in vivo rat model. Cancer Immunol Immunother 67, 67–77 (2018). https://doi.org/10.1007/s00262-017-2063-2

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  • DOI: https://doi.org/10.1007/s00262-017-2063-2

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