Comparative Study of Extracellular Vesicles from the Urine of Healthy Individuals and Prostate Cancer Patients.
Recent studies suggest that extracellular vesicles may be the key to timely diagnosis and monitoring of genito-urological malignancies. In this study we investigated the composition and content of extracellular vesicles found in the urine of healthy donors and prostate cancer patients. Urine of 14 PCa patients and 20 healthy volunteers was clarified by low-speed centrifugation and total extracellular vesicles fraction was obtain by high-speed centrifugation. The exosome-enriched fraction was obtained by filtration of total extracellular vesicles through a 0.1 μm pore filter. Transmission electron microscopy showed that cell-free urine in both groups contained vesicles from 20 to 230 nm. Immunogold staining after ultrafiltration demonstrated that 95% and 90% of extracellular vesicles in healthy individuals and cancer patients, respectively, were exosomes. Protein, DNA and RNA concentrations as well as size distribution of extracellular vesicles in both fractions were analyzed. Only 75% of the total protein content of extracellular vesicles was associated with exosomes which amounted to 90-95% of all vesicles. Median DNA concentrations in total extracellular vesicles and exosome-enriched fractions were 18 pg/ml and 2.6 pg/ml urine, correspondingly. Urine extracellular vesicles carried a population of RNA molecules 25 nt to 200 nt in concentration of no more than 290 pg/ml of urine. Additionally, concentrations of miR-19b, miR-25, miR-125b, and miR-205 were quantified by qRT-PCR. MiRNAs were shown to be differently distributed between different fractions of extracellular vesicles. Detection of miR-19b versus miR-16 in total vesicles and exosome-enriched fractions achieved 100%/93% and 95%/79% specificity/sensitivity in distinguishing cancer patients from healthy individuals, respectively, demonstrating the diagnostic value of urine extracellular vesicles.
Laboratory of molecular medicine, Institute of Chemical Biology and Fundamental Medicine CD RAS, Novosibirsk, Russian Federation.
Olga E
OE
Bryzgunova
Dispensary department 2, Novosibirsk Regional Oncology Center, Novosibirsk, Russian Federation.
Marat M
MM
Zaripov
Laboratory of molecular medicine, Institute of Chemical Biology and Fundamental Medicine CD RAS, Novosibirsk, Russian Federation.
Tatyana E
TE
Skvortsova
Laboratory of molecular medicine, Institute of Chemical Biology and Fundamental Medicine CD RAS, Novosibirsk, Russian Federation.
Evgeny A
EA
Lekchnov
Group of microscopy, Institute of Chemical Biology and Fundamental Medicine CD RAS, Novosibirsk, Russian Federation.
Alina E
AE
Grigor'eva
Laboratory of molecular medicine, Institute of Chemical Biology and Fundamental Medicine CD RAS, Novosibirsk, Russian Federation.
Ivan A
http://orcid.org/0000-0001-9101-2207
IA
Zaporozhchenko
Laboratory of molecular medicine, Institute of Chemical Biology and Fundamental Medicine CD RAS, Novosibirsk, Russian Federation.|Centre of Oncology and Radiotherapy, Novosibirsk Research Institute of Circulation Pathology Academician E.N. Meshalkin, Novosibirsk, Russian Federation.
Evgeny S
ES
Morozkin
Group of microscopy, Institute of Chemical Biology and Fundamental Medicine CD RAS, Novosibirsk, Russian Federation.
Elena I
EI
Ryabchikova
Center of New Medical Technologies of ICBFM SB RAS, Novosibirsk, Russian Federation.
Yuri B
YB
Yurchenko
Dispensary department 2, Novosibirsk Regional Oncology Center, Novosibirsk, Russian Federation.
Vladimir E
VE
Voitsitskiy
Laboratory of molecular medicine, Institute of Chemical Biology and Fundamental Medicine CD RAS, Novosibirsk, Russian Federation.|Centre of Oncology and Radiotherapy, Novosibirsk Research Institute of Circulation Pathology Academician E.N. Meshalkin, Novosibirsk, Russian Federation.
Pavel P
PP
Laktionov
D000368:Aged; D000369:Aged, 80 and over; D002498:Centrifugation / Q000379:methods; D004273:DNA, Neoplasm / Q000235:genetics / Q000652:urine; D055354:Exosomes / Q000378:metabolism* / Q000648:ultrastructure; D000067128:Extracellular Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000652:urine; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000378:metabolism / Q000652:urine; D011471:Prostatic Neoplasms / Q000175:diagnosis / Q000652:urine*; D012334:RNA, Neoplasm / Q000235:genetics / Q000652:urine; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D004273:DNA, Neoplasm; C505362:MIRN16 microRNA, human; C549214:MIRN19A microRNA, human; D035683:MicroRNAs; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm