Extracellular plasma RNA from colon cancer patients is confined in a vesicle-like structure and is mRNA-enriched
Open accessAbstract
Little is yet known about the origin and protective mechanism of free nucleic acids in plasma. We investigated the possibility of these free nucleic acids being particle associated. Plasma samples from colon cancer patients and cell culture media were subjected to various antibody incubations, ultracentrifugation, and RNA extraction protocols for total RNA, epithelial RNA, and mRNA. Flow cytometry using a Ber-EP4 antibody and confocal laser microscopy after staining with propidium iodide were also performed. mRNA levels of the LISCH7 and SDHA genes were determined in cells and in culture media. Ber-EP4 antibody and polystyrene beads coated with oligo dT sequences were employed. We observed that, after incubation, total RNA and mRNA were always detected after membrane digestion, and that epithelial RNA was detected before this procedure. In ultracentrifugation, mRNA was caught in the supernatant only if a former lysis mediated or in the pellet if there was no previous digestion. Flow cytometry determinations showed that antibody-coated microbeads keep acellular structures bearing epithelial antigens apart. Confocal laser microscopy made 1- to 2-μm-diameter particles perceptible in the vicinity of magnetic polystyrene beads. Relevant differences were observed between mRNA of cells and culture media, as there was a considerable difference in LISCH7 mRNA levels between HT29 and IMR90 cell co-cultures and their culture media. Our results support the view that extracellular RNA found in plasma from cancer patients circulates in association with or is protected in a multiparticle complex, and that an active release mechanism by tumor cells may be a possible origin.
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Owing to the fact that the extracellular compartment contains RNases, the estimation of circulating hTERT transcripts is unreliable due to mRNA digestion.
“However, owing to the fact that the extracellular compartment contains RNases, the estimation of circulating hTERT transcripts is unreliable due to mRNA digestion [211].”
From Current and Future Trends of Colorectal Cancer Treatment: Exploring Advances in Immunotherapy (2024) · cites this paper as [211]
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- Authors
- García, J. M., García, V., Peña, C., Domínguez, G., Silva, J., Diaz, R., Espinosa, P., Citores, M. J., Collado, M., & Bonilla, F.
- Affiliations
- Hospital Universitario Puerta de Hierro Majadahonda (ES)
- Spanish National Cancer Research Centre (ES)
- Venue
- RNA
- Publisher
- Cold Spring Harbor Laboratory
- Published
- 2008-05-02
- Volume
- 14
- Issue
- 7
- Pages
- 1424-1432
- ISSN
- 1355-8382 (ISSN-L)
- 1469-9001
- PMC
- PMC2441977
- OpenAlex
- W2117089920
- Cited by
- 86
- References
- 41
- Citations by year
- 2025: 1 · 2024: 1 · 2023: 1 · 2022: 4 · 2020: 2 · 2019: 5 · 2018: 5 · 2017: 5 · 2016: 4 · 2015: 6 · 2014: 13 · 2013: 6 · 2012: 11
- FWCI
- 3.01
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- RNA (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- HighWire Press Open Archive (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer Genomics and Diagnostics
- Keywords
- extracellular RNA, plasma RNA, colon cancer, vesicle-like structures, mRNA enrichment, SDHA, extracellular vesicles, HT-29 cells, IMR-90 cells
- MeSH terms
- Adenocarcinoma, Colonic Neoplasms, Fibroblasts, Humans, Neoplastic Cells, Circulating, RNA, Messenger, RNA, Neoplasm, Cell Line, Tumor
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