MiR-21-5p in urinary extracellular vesicles is a novel biomarker of urothelial carcinoma.
BACKGROUND
Extracellular vesicles are lipid bilayer vesicles containing protein, messengerRNA and microRNA. Cancer cell-derived extracellular vesicles may be diagnostic and therapeutic targets. We extracted extracellular vesicles from urine of urothelial carcinoma patients and the control group to identify cancer-specific microRNAs in urinary extracellular vesicles as new biomarkers.


METHODS
microRNA from urinary extracellular vesicles extracted from 6 urothelial carcinoma patients and 3 healthy volunteers was analyzed. We verified candidate microRNAs in an independent cohort of 60 urinary extracellular vesicles samples. To normalize the microRNA expression level in extracellular vesicles, we examined the following in extracellular vesicles: protein concentration, CD9 intensity, amounts of whole miRNAs, RNA U6B small nuclear expression and the creatinine concentration of original urine correlating with the counts of extracted extracellular vesicles measured by the NanoSight™ system.


RESULTS
From the microarray results 5 microRNAs overexpressed in urinary extracellular vesicles of urothelial carcinoma patients were identified. Creatinine concentration of original urine correlated most with particle counts of extracellular vesicles, indicating that creatinine could be a new tool for normalizing microRNA expression. MiR-21-5p was the most potent biomarker in urinary extracellular vesicles (sensitivity, 75.0%; specificity, 95.8%) and was also overexpressed in urinary extracellular vesicles from urothelial carcinoma patients with negative urine cytology. For the subgroup with negative urine cytology, the sensitivity was 75.0% and specificity was 95.8%.


CONCLUSIONS
MiR-21-5p in urinary extracellular vesicles could be a new biomarker of urothelial carcinoma, especially for urothelial carcinoma patients with negative urine cytology.
Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.
Kyosuke
K
Matsuzaki
Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.
Kazutoshi
K
Fujita
Laboratory of Molecular and Cellular Physiology, Osaka University Graduate School of Pharmaceutical Science, Osaka, Japan.
Kentaro
K
Jingushi
Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.
Atsunari
A
Kawashima
Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.
Takeshi
T
Ujike
Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.
Akira
A
Nagahara
Laboratory of Molecular and Cellular Physiology, Osaka University Graduate School of Pharmaceutical Science, Osaka, Japan.
Yuko
Y
Ueda
Department of Urology, Osaka General Medical Center, Osaka, Japan.
Go
G
Tanigawa
Department of Urology, Osaka Police Hospital, Osaka, Japan.
Iwao
I
Yoshioka
Cancer Proteomics Group, Genome Center, Japanese Foundation for Cancer Research, Tokyo, Japan.
Koji
K
Ueda
Department of Immunology, Kanazawa University Graduate School of Medical Sciences, Ishikawa, Japan.
Rikinari
R
Hanayama
Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.
Motohide
M
Uemura
Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.
Yasushi
Y
Miyagawa
Laboratory of Molecular and Cellular Physiology, Osaka University Graduate School of Pharmaceutical Science, Osaka, Japan.
Kazutake
K
Tsujikawa
Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.
Norio
N
Nonomura
D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000652:urine*; D000067128:Extracellular Vesicles / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000652:urine*; D008875:Middle Aged; D014571:Urologic Neoplasms / Q000235:genetics / Q000473:pathology / Q000652:urine*
D014408:Biomarkers, Tumor; C524940:MIRN21 microRNA, human; D035683:MicroRNAs
biomarker; extracellular vesicles; microRNA; urine; urothelial carcinoma