Abstract
Extracellular vesicles (EVs) are shed by many different cell types. Their nucleic acids content offers new opportunities for biomarker research in different solid tumors. The role of EV RNA in prostate cancer (PCa) is still largely unknown. EVs were isolated from different benign and malignant prostate cell lines and blood plasma from patients with PCa (n = 18) and controls with benign prostatic hyperplasia (BPH) (n = 7). Nanoparticle tracking analysis (NTA), Western blot, electron microscopy, and flow cytometry analysis were used for the characterization of EVs. Non-coding RNA expression profiling of PC3 metastatic PCa cells and their EVs was performed by next generation sequencing (NGS). miRNAs differentially expressed in PC3 EVs were validated with qRT-PCR in EVs derived from additional cell lines and patient plasma and from matched tissue samples. 92 miRNAs were enriched and 48 miRNAs were depleted in PC3 EVs compared to PC3 cells, which could be confirmed by qRT-PCR. miR-99b-5p was significantly higher expressed in malignant compared to benign EVs. Furthermore, expression profiling showed miR-10a-5p (p = 0.018) and miR-29b-3p (p = 0.002), but not miR-99b-5p, to be overexpressed in plasma-derived EVs from patients with PCa compared with controls. In the corresponding tissue samples, no significant differences in the miRNA expression could be observed. We thus propose that EV-associated miR-10a-5p and miR-29b-3p could serve as potential new PCa detection markers.
Highlights
Extracellular vesicles (EVs) are defined as particles delimited by a lipid bilayer that cannot replicate and can be subclassified according to their size, density, biochemical composition, and origin [1]
We used next generation sequencing (NGS) profiling of small ncRNA to identify 92 miRNAs that were enriched and 48 miRNAs that were depleted in PC3 EVs
The largest group of miRNAs that appeared to be enriched in EVs was the miR-10 family
Summary
Extracellular vesicles (EVs) are defined as particles delimited by a lipid bilayer that cannot replicate and can be subclassified according to their size, density, biochemical composition, and origin [1]. Active surface proteins give them antigenic potency, capable of both inducing immune response and interacting with cellular surface structures [4,5,6] When taken up, their biologically active contents are released, enabling various effects on transcription, translation, and post-translational modifications, leading to significant changes in the metabolism and homeostasis of the recipient cell [7,8,9]. Their biologically active contents are released, enabling various effects on transcription, translation, and post-translational modifications, leading to significant changes in the metabolism and homeostasis of the recipient cell [7,8,9] Apart from their physiological functions, for instance, in immune-modulation, angiogenesis, and proliferation, they are crucial to many pathophysiological processes [10,11], like the modification of the tumor microenvironment in the paracrine setting [12,13] or pre-metastatic niche formation via the blood stream [14]. Small EVs in the size range between 30 and 100 nm are termed as exosomes, though a clear differentiation from other EVs regarding the composition and function is not possible [1]
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