Screening and validation of novel serum panel of microRNA in stratification of prostate cancer

Abstract

Background: Owing to the heterogeneous nature of prostate cancer (PCa) and errors in the characterization of the disease, researchers have been trying to unveil molecular biomarkers like microRNA (miRNA) as diagnostic markers. The purpose of our study is to demonstrate the precision of a panel of miRNAs as biomarkers with diagnostic potential for risk stratification.

Materials and methods: The present study demonstrates the comparative expression profiles of miRNA-141,-1290,-100, and -335 in both tissue and serum, including Benign Prostate Hyperplasia (BPH) and PCa, with healthy volunteers. Firstly, we demonstrate the expression of all miRNAs in the discovery cohort, including metastasis and benign tissue, and later validate their non-invasive diagnostic potential in BPH and PCa with healthy volunteers. MiRNA was isolated from tissue and serum to be quantified by RT-PCR and analyzed for biomarker potential by receiver operating characteristic (ROC) curve analysis, followed by targetome analysis of each miRNA.

Results: Among the non-invasive miRNA assessed, it was seen that miRNA 141 (P = 0.0003) and miRNA 1290 (P < 0.0001) are oncogenic with significantly higher expression, while miRNA 100 (P = 0.0002) and miRNA 335 are tumor suppressor, in PCa as compared to controls. While for BPH, miRNA 141 (P = 0.003) and miRNA 335 (P = 0.0002) were found to be significantly oncogenic and tumor suppressors, respectively. The analysis of the ROC curve of panel miRNAs (miRNA-141,-1290, and -100) portrayed a significant area under the curve with greater sensitivity and specificity. Moreover, in-silico prediction of their respective targetomes represents their extensive involvement in PCa progression and various other cascades that aid in PCa networks.

Conclusions: To the best of our knowledge, we are going to report for the first time this panel of miRNA that can be used to accurately and efficiently diagnose BPH and PCa patients from healthy males.

Keywords: Benign prostatic hyperplasia; Biomarker; Diagnosis; MicroRNA; Non-invasive; Prostate cancer.

Fig. 1

Study design.

Fig. 2

The quantitative expression analysis of miRNA 141, miRNA 100, miRNA 335, and miRNA 1290 between mPCa and the control group in the screening cohort.

Fig. 3

Box plot for the quantitative expression of (A) miRNA 141, (B) miRNA 100, (C) miRNA 335, and (D) miRNA 1290 in the serum of BPH and PCa patients.

Fig. 4

ROC curve analysis of individual (A) miRNA 141 (B) miRNA 100 (C) miRNA 335 (D) miRNA 1290 for BPH and PCa groups.

Fig. 5

ROC curve analysis of combination (A) all four miRNAs in both groups (B) signature panel of three, miRNA 100, miRNA 141, and miRNA 1290 for PCa group (C) panel of miRNA 335 and miRNA 141 for BPH group.

Fig. 6

(A) Comparative analysis between intragroup PSA levels (Low PSA – below 4 ng/ml; Gray PSA level; 4–10 ng/ml; High PSA level-above 10 ng/ml) in the Prostate Cancer group. (B): Comparative analysis between intragroup Gleason Score (GS) (GS = 7; GS > 7) in the prostate cancer group.

Fig. 7

KEGG pathway enrichment analysis of genes targeted by (A) the PCa panel of miRNA (B) the BPH panel of miRNA.

Fig. 8

Cytoscape 3.9.1: String app-the interaction of molecular targets of panel miRNA involved in PCa with confidence level >0.7 and an enrichment score of 2.55E-15.