The Panel of 12 Cell-Free MicroRNAs as Potential Biomarkers in Prostate Neoplasms

Abstract

Prostate cancer is a global biological, medical, and social issue aggravated by the lack of reliable, highly specific, and sensitive non-invasive tests for diagnosis and staging of prostate cancer. One prospective source of biomarkers are the cell-free miRNAs present in various biological fluids. In the present study, we validated the diagnostic potential of cell-free miRNAs: miR-19b, miR-22, miR-92a, miR-378, miR-425, miR-30e, miR-31, miR-125b, miR-200b, miR-205, miR-375, and miR-660; we estimated the required sample size and the minimal miRNA set for a subsequent large-scale validation study. Relative expression of 12 miRNA combined in 31 ratios was investigated in three fractions of biological fluids (urine extracellular vesicles, clarified urine, and plasma) obtained from patients with prostate cancer (n = 10), benign prostate hyperplasia (n = 8), and healthy volunteers (n = 11). Eight of the miRNAs found in urine vesicles (miR-19b, miR-30e, miR-31, miR-92a, miR-125, miR-200, miR-205, and miR-660) showed great promise and when combined into six ratios (miR-125b/miR-30e, miR-200/miR-30e, miR-205/miR-30e, miR-31/miR-30e, miR-660/miR-30e, and miR-19b/miR-92a) could classify patients with prostate cancer, benign prostate hyperplasia, and healthy donors with 100% specificity, 100% sensitivity, and with a high degree of reliability for most donors.

Total Blood Exosomes in Breast Cancer: Potential Role in Crucial Steps of Tumorigenesis

“…Each RNA sample was reverse-transcribed to cDNA using the TaqMan ® MicroRNA Reverse Transcriptions Kit (PN 4366596, Applied Biosystems, USA). Single-stranded cDNA was synthesized from 2.5 μL of RNA using specific miRNA primers [ 31 , 47 , 48 ]. Samples without RNA templates were used as negative controls.…”

“…A total of 4 μL of cDNA was used as a template in a 24 μL PCR reaction. PCR products were amplified using specific primers [ 31 , 47 , 48 ] and TaqMan Universal PCR Master Mix (PN 4427788, Applied Biosystems, USA) and detected using the CFX 96TM Real-Time System (Bio-Rad, USA). The PCR reactions for each sample were run in duplicate, including blank controls without cDNA.…”

The Influence of Radical Prostatectomy on the Expression of Cell-Free MiRNA

“…Primers and probes for reverse transcription and TaqMan qPCR [ 15 ] were synthesized in the Laboratory of Medicinal Chemistry (ICBFM SB RAS, Novosibirsk, Russia), their sequences are presented in Table S1 . Each reverse transcription (RT) reaction was performed in a total volume of 10 μL and contained 2.5 μL of RNA, 25-nM of each of the miRNA-specific primers, 0.5 units of RiboLockRNAse inhibitor (Fermentas, Vilnius, Lithuania), 50 units of M-MuLV–RH reverse transcriptase (BiolabMix, Novosibirsk, Russia), 2 μL of 5 × MMLV reaction buffer (250-mM Tris-HCl (pH 8.3 at 25 °C), 250-mM KCl, 20-mM MgCl 2 and 50-mM DTT) and 125-mM of each dNTP.…”

“…Earlier, we demonstrated that, selected from 84 miRNAs [ 14 ], a panel of 12 cell-free miRNAs—namely, miR-19b, miR-22, miR-92a, miR-378, miR-425, miR-30e, miR-31, miR-125b, miR-200b, miR-205, miR-375 and miR-660—allows us to classify patients with PCa and benign prostate hyperplasia (BPH) against healthy donors (HD) with 100% specificity and 100% sensitivity [ 15 ].…”

The effects of mutant Ras proteins on the cell signalome

“…It was recently observed that both the miR-21 and K-Ras expression levels were elevated in colorectal cancer samples, suggesting that, instead of suppressing the Ras pathway, miR-21 may promote K-Ras mRNA translation during cancer development [116]. Over the last decade, the pleiotropic effects and biological significance of microRNAs in Ras signaling modulation have been established, and the use of microRNA levels as biomarkers for cancer diagnosis is under extensive research [117,118].…”