Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

Search BRD Papers

Investigating differential miRNA expression profiling using serum and urine specimens for detecting potential biomarker for early prostate cancer diagnosis.

Author(s): HasanoĞlu S, GÖncÜ BS, YÜcesan E, Atasoy S, Kayali Y, Özten KandaŞ N

Publication: Turk J Med Sci, 2021, Vol. , Page

PubMed ID: 33550766 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study compared the RNA yield and miRNA expression profile of a pooled urine and serum specimen from prostate cancer patients and healthy controls. Levels of miR-320A and miR-6750 were further investigated in individual plasma and serum specimens from prostate cancer patients and healthy controls. Urine and EDTA blood were collected from eight prostate cancer patients and 30 healthy patients. Serum was obtained by centrifugation of the blood at 1100 x g for 10 min and frozen at -80°C. Serum was thawed on ice and RNA was extracted using Trizol reagent and stored at -80°C. Urine was centrifuged at 1210 x g for 20 minutes and the pellet was resuspended in PBS. RNA was extracted from urine using the Urine Total RNA Purification and stored frozen. RNA was quantified by spectrophotometer. RNA expression was analyzed by applying pooled extracted RNA from urine and serum to an Affymetrix GeneChip miRNA 4.0 Array. Levels of miR-320A and miR-6750 were quantified by real-time RT-PCR in individual urine and serum specimens using the miScript II RT Kit RT-PCR Kit.

   Summary of Findings:

   The median RNA yield from urine and serum of healthy patients was 507.7 ng/μL and 23.2 ng/μL, respectively, and from prostate cancer patients was 385.3 ng/μL and 22.71 ng/μL, respectively. A total of 103 miRNAs were expressed in all four pools (serum and urine of prostate cancer patients and healthy controls). Of the 103 miRNAs identified, 49 were higher and 14 were lower in the pooled urine from cancer patients than the pooled urine from healthy controls. In serum, 19 miRNAs were higher and 21 were lower in the pool from cancer patients than the pool from healthy controls. Four miRNAs were found to differ (>2-fold change) in the pooled serum and urine from cancer patients and healthy controls. Two of these (mIR-4532 and miR-4706) were higher in the urine (3.99- and 2.66-fold, respectively) but lower in the serum (-2.05- and -2.72-fold, respectively) of cancer patients than controls. miR-6750 was found to be higher in both the pooled serum and urine from cancer patients compared to healthy controls (2.37- and 2.23-fold, respectively). In contrast, miR-320A was lower in the pooled serum and urine of cancer patients than controls (-2.05 and -4.2-fold, respectively). Real-time RT-PCR found significantly higher levels of miR-320A in serum than urine (P=0.0023 in control patients and P=0.0026 in cancer patients) and, in contrast to the microarray results, higher levels in the urine but not serum of cancer patients than controls (P=0.0168). miR-6750 was higher in the serum of cancer patients than urine (P=0.007) but levels did not differ between cancer patients and controls in either urine or serum.

   Biospecimens

   • Bodily Fluid - Urine
   • Bodily Fluid - Serum

   Preservative Types

   • None (Fresh)
   • Frozen

   Diagnoses:

   • Neoplastic - Carcinoma
   • Normal

   Platform:

   Analyte	Technology Platform
   RNA	Spectrophotometry
   RNA	DNA microarray
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Preaquisition	Diagnosis/ patient condition	Prostate cancer Healthy
   Real-time qRT-PCR Specific	Targeted nucleic acid	miR-6750 miR-320A
   Biospecimen Acquisition	Biospecimen location	Serum Urine

   View more