Integrative functional genetic-epigenetic approach for selecting genes as urine biomarkers for bladder cancer diagnosis
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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.
KeywordsBladder cancer Urine cytology Long non-coding RNA-UCA1 Bilharziasis miRNA Hyaluronoglucosaminidase 1 Urinary biomarkers
Long non-coding RNA
Quantitative polymerase chain reaction
Reverse transcription polymerase chain reaction
Squamous cell carcinoma
Transitional cell carcinoma
Urothelial cancer associated 1
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
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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