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Integrative functional genetic-epigenetic approach for selecting genes as urine biomarkers for bladder cancer diagnosis

  • Research Article
  • Published:
Tumor Biology

Abstract

Early screening for bladder cancer (BC) holds the key to combat and control the increasing global burden of BC mortality. We presented a simple approach to characterize, analyze, and validate a panel of biomarkers in BC and their relationship to bilharziasis. We investigated voided urine and blood samples from patients with bladder cancer (n = 94), benign bladder lesions (n = 60), and age-matched normal controls (n = 56). This study was divided into the following phases. (1) We analyzed the expression of urinary Hyaluronoglucosaminidase 1 (HYAL1) protein in BC and control samples by zymography. (2) We performed bioinformatics analysis to retrieve a set of epigenetic regulators of HYAL1. (3) This set of three selected genes [long non-coding RNA-urothelial cancer associated 1(lncRNA-UCA1), microRNA-210, and microRNA-96] was then analyzed in the same urine samples used in phase I by quantitative real-time PCR. (4) A high reproducibility of gene selection results was also determined from statistical validation. The urinary expression of HYAL1 protein and its epigenetic regulators were higher in BC patients (P < .001). The receiver-operating characteristic curve analyses demonstrated that each one had good sensitivity and specificity for distinguishing BC patients from non-BC ones (HYAL1, 89.4 and 91.2 %; miR-210, 76.6 and 93 %; miR-96, 76.6 and 89.4 %; and lncRNA-UCA1, 91.5 and 96.5 %). There was a significant positive correlation between HYAL1 and the selected epigenetic biomarkers. The performance of this urine biomarker panel reached 100 % sensitivity and 89.5 % specificity for bladder cancer diagnosis.

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Abbreviations

BC:

Bladder cancer

HYAL1:

Hyaluronoglucosaminidase 1

kDa:

Kilodalton

LncRNA:

Long non-coding RNA

MiR:

Micro-RNA

qPCR:

Quantitative polymerase chain reaction

RT-PCR:

Reverse transcription polymerase chain reaction

SCC:

Squamous cell carcinoma

TCC:

Transitional cell carcinoma

UCA1:

Urothelial cancer associated 1

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Acknowledgments

This work was supported by Ain Shams University Research Projects 2013–14. The authors are grateful to Dr. Nahla M. Awad, Ass. Prof. of Pathology at Early Cancer Detection Unit, Faculty of Medicine, Ain Shams University, for her help in the cytological examinations of all investigated urine samples. All authors have read the journal’s policy on disclosure of potential conflicts of interest. The authors have no conflict of interest. All authors have read the journal’s authorship agreement and that the manuscript has been reviewed by and approved by all named authors.

Conflicts of interest

None.

Authors’ contributions

Eissa S has participated in the design of the study, carried out data analysis, involved in drafting the manuscript or revising and has given final approval of the version to be published. Matboli M has performed bioinformatic analysis, practical work, participated in the design of the study, and performed the statistical analysis. Essawy N participated in the study design and involved in drafting the manuscript or revising. Youssef M. Kotb has provided us with urine, blood samples, and patient data and has given final approval of the version to be published. All authors read and approved the final manuscript.

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Authors and Affiliations

  1. Oncology Diagnostic Unit, Medical Biochemistry and Molecular biology Department, Faculty of Medicine, Ain Shams University, P.O. box 11381, Abbassia, Cairo, Egypt

    Sanaa Eissa & Marwa Matboli

  2. Pharmacogenetics and Stratified Medicine, University College London, London, UK

    Nada O. E. Essawy

  3. Urology Department, Faculty of Medicine, Ain Shams University, Cairo, Egypt

    Youssef M. Kotb

Authors
  1. Sanaa Eissa
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  2. Marwa Matboli
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  3. Nada O. E. Essawy
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  4. Youssef M. Kotb
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Correspondence to Sanaa Eissa.

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Eissa, S., Matboli, M., Essawy, N.O.E. et al. Integrative functional genetic-epigenetic approach for selecting genes as urine biomarkers for bladder cancer diagnosis. Tumor Biol. 36, 9545–9552 (2015). https://doi.org/10.1007/s13277-015-3722-6

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  • DOI: https://doi.org/10.1007/s13277-015-3722-6

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