Direct quantitative detection for cell-free miR-155 in urine: a potential role in diagnosis and prognosis for non-muscle invasive bladder cancer
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Xin Zhang1, Yanli Zhang2, Xinfeng Liu2, Aiju Fang3, Jinfeng Wang1, Yongmei Yang1, Lili Wang1, Lutao Du1, Chuanxin Wang1
1Department of Clinical Laboratory, Qilu Hospital, Shandong University, Jinan 250012, China
2Department of Clinical Laboratory, Traffic Hospital of Shandong Province, Jinan 250031, China
3Department of Pathology, Traffic Hospital of Shandong Province, Jinan 250031, China
Chuanxin Wang, e-mail: email@example.com
Keywords: cell-free, miR-155, urine, non-muscle invasive bladder cancer, biomarker
Received: July 07, 2015 Accepted: November 21, 2015 Published: December 02, 2015
High recurrence rates of non-muscle invasive bladder cancer (NMIBC) in patients require lifelong testing and monitoring. The aim of this study is to develop a simplified RT-qPCR method (RT-qPCR-D) which directly quantifies cell-free miR-155 in urine without RNA extraction, and assess it as a potential tool in NMIBC detection. A pilot study including 60 urine samples was used to investigate the feasibility of RT-qPCR-D in detecting cell-free miR-155. Then, miR-155 levels were quantified in a large independent cohort of urine from 162 NIMBC patients, 76 cystitis patients, and 86 healthy donors using the RT-qPCR-D method. Changes of cell-free miR-155 before and after operation were also analyzed in 32 NIMBC patients. In pilot study, we found a significant linear association between RT-qPCR and RT-qPCR-D in urinary miR-155 detection. Both methods showed cell-free miR-155 were significantly increased in NMIBC patients, and could reflect their expression in tissues. Then, the increased expression of cell-free miR-155 was successfully validated in 162 NIMBC patients when compared with cystitis patients and healthy donors. Moreover, it distinguished NMIBC patients from others with 80.2% sensitivity and 84.6% specificity, which was superior to urine cytology. Cell-free miR-155 correlated with NMIBC stage and grade, and was an independent factor for predicting recurrence and progression to muscle invasion. In addition, cell-free miR-155 was significantly decreased after NMIBC patients underwent transurethral bladder resection. In conclusion, detection of cell-free miR-155 in urine using RT-qPCR-D is a simple and noninvasive approach which may be used for NMIBC diagnosis and prognosis prediction.
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