Establishment of urinary micro-RNA detection method and its application in diagnosis of bladder cancer

doi:10.16150/j.1671-2870.2017.01.018

Abstract

Abstract: Objective: To establish a method for detection of 8 urinary micro-RNA and to detect their distribution in urine and evaluating its significance in detection of bladder cancer. Methods: Early morning and random urine samples from 15 healthy controls and random urine samples from 15 patients with bladder cancer were collected. The total RNA were extracted and then were reversely transcripted to cDNA, and 8 micro-RNAs (miR-126, miR-200a, miR-200b, miR-200c, miR-182, miR-183, miR-429 and miR-141) were amplified with real-time polymerase chain reaction (PCR). Diffe-rences in miRNA levels were compared between early morning and random urine samples as well as between non-concentrated and concentrated samples. Different extraction and purification methods for miRNAs were compared and the diffe-rences of miRNA concentrations between controls and bladder cancer patients were analyzed. The diagnostic efficacy of multiple urinary miRNA combinations for bladder cancer was analyzed using ROC curve. Results: The relative concentrations of 6 miRNAs (miR-126, miR-200a, miR-200b, miR-200c, miR-429 and miR-141) were significantly higher in the concentrated urine than those in non-concentrated urine. There was no significant difference in relative miRNA concentration between early morning and randomized urine collection. Compared with normal controls, the relative concentrations of 4 miRNAs (miR-126, miR-182, miR-183 and miR-141) were significantly increased in bladder cancer group while miRNA-200a was significantly decreased. ROC analysis showed that detection of single miRNA had limited diagnostic efficacy for bladder cancer, while combined analysis of 5 miRNA had better diagnostic value for bladder cancer. Conclusions: Multiple micro-RNAs are stably distributed in urine, and combined detection of multiple miRNAs might be used as a dia-gnostic marker for bladder cancer.

Key words: Bladder cancer, MicroRNA, Urine, Real-time polymerase chain reaction

CLC Number:

R737.14

LI Li, BIAN Bingxian, ZHANG Liang, SHEN Lisong. Establishment of urinary micro-RNA detection method and its application in diagnosis of bladder cancer[J]. Journal of Diagnostics Concepts & Practice, 2017, 16(01): 93-97.

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