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Updated: May 18, 2026
Sequencing Small Non-coding RNA from Formalin-fixed Tissues and Serum-derived Exosomes from Castration-resistant Prostate Cancer Patients
Published on: November 19, 2019
Profiling of microRNAs in exosomes released from PC-3 prostate cancer cells
Nina Pettersen Hessvik1, Santosh Phuyal, Andreas Brech
1Department of Biochemistry, The Norwegian Radium Hospital, Oslo, Norway.
Abstract:
Exosomes are small extracellular vesicles released to the extracellular milieu through fusion of multivesicular bodies with the plasma membrane. These vesicles contain microRNAs and might therefore be vehicles transferring genetic information between cells. The aim of this study was to investigate whether there was a sorting of microRNAs into exosomes in the prostate cancer cell line PC-3. In addition, microRNAs in PC-3 cells and in the non-cancerous prostate cell line RWPE-1 were compared. Exosomes were isolated from the conditioned media from PC-3 cells by ultracentrifugation and inspected by electron microscopy. Total RNA was isolated and microRNAs were analyzed by microarray analysis and real time RT-PCR. MicroRNA microarray analysis revealed that the microRNA profile of PC-3 released exosomes was similar to the profile of the corresponding parent cells. Nevertheless, a sorting of certain microRNAs into exosomes was observed, and low number microRNAs (microRNAs with a low number in their name) were found to be underrepresented in these vesicles. Moreover, the miRNA profile of PC-3 cells resembled the miRNA profile of RWPE-1 cells, though some miRNAs were found to be differently expressed in these cell lines. These results show that exosomes from PC-3 cells, in agreement with previous reports from other cell types, contain microRNAs. Furthermore, this study supports the idea that there is a sorting of microRNAs into exosomes and adds a new perspective by pointing at the underrepresentation of low number miRNAs in PC-3 released exosomes.
Insights
Prostate cancer cells release exosomes containing microRNAs, with specific sorting observed. Low-number microRNAs are underrepresented in these exosomes, offering new insights into exosome cargo selection.
Area of Science:
- Cell Biology
- Molecular Biology
- Cancer Research
Background:
- Exosomes are extracellular vesicles involved in intercellular communication.
- MicroRNAs within exosomes may transfer genetic information between cells.
- Understanding exosome cargo is crucial for cancer research.
Purpose of the Study:
- To investigate microRNA sorting into exosomes from the PC-3 prostate cancer cell line.
- To compare microRNA profiles between PC-3 cells and non-cancerous RWPE-1 cells.
- To identify specific microRNAs enriched or depleted in PC-3 exosomes.
Main Methods:
- Isolation of exosomes from PC-3 cell conditioned media via ultracentrifugation.
- Electron microscopy for exosome characterization.
- Microarray analysis and real-time RT-PCR for microRNA profiling.
Main Results:
- PC-3 cell-derived exosome microRNA profiles mirror their parent cells.
- Specific microRNA sorting into exosomes was confirmed.
- Low-number microRNAs were found to be underrepresented in PC-3 exosomes.
- PC-3 and RWPE-1 cell microRNA profiles showed similarities with some differential expression.
Conclusions:
- PC-3 cell exosomes contain microRNAs, supporting their role in cargo transport.
- Evidence suggests active sorting of microRNAs into exosomes.
- The underrepresentation of low-number microRNAs in PC-3 exosomes provides novel insights into cargo selection mechanisms.
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