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| Status |
Public on Oct 12, 2016 |
| Title |
Real-time quantitative PCR analysis of shed microvesicles and exosomes released by human colorectal cancer LIM1863 cell lines |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by RT-PCR
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| Summary |
Secreted microRNAs (miRNAs) enclosed within extracellular vesicles (EVs) play a pivotal role in intercellular communication by regulating recipient cell gene expression and affecting target cell function. Here, we report the isolation of three distinct EV subtypes from the human colon carcinoma cell line LIM1863--shed microvesicles (sMVs) and two exosome populations (immunoaffinity isolated A33-exosomes and EpCAM-exosomes). Deep sequencing of miRNA libraries prepared from parental LIM1863 cells/derived EV subtype RNA yielded 254 miRNA identifications, of which 63 are selectively enriched in the EVs--miR-19a/b-3p, miR-378a/c/d, and miR-577 and members of the let-7 and miR-8 families being the most prominent. Let-7a-3p*, let-7f-1-3p*, miR-451a, miR-574-5p*, miR-4454 and miR-7641 are common to all EV subtypes, and 6 miRNAs (miR-320a/b/c/d, miR-221-3p, and miR-200c-3p) discern LIM1863 exosomes from sMVs; miR-98-5p was selectively represented only in sMVs. Notably, A33-Exos contained the largest number (32) of exclusively-enriched miRNAs; 14 of these miRNAs have not been reported in the context of CRC tissue/biofluid analyses and warrant further examination as potential diagnostic markers of CRC. Surprisingly, miRNA passenger strands (star miRNAs) for miR-3613-3p*, -362-3p*, -625-3p*, -6842-3p* were the dominant strand in A33-Exos, the converse to that observed in parental cells. This finding suggests miRNA biogenesis may be interlinked with endosomal/exosomal processing.
This work was supported by National Health and Medical Research Council of Australia (NHMRC) program grant #487922.
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| Overall design |
qRT-PCR miRNA expression profiling. Same amount of total RNA was used for LIM1863 cells and released extracellular vesicles (sMVs, A33-Exos and EpCAM-Exos).
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| Contributor(s) |
Chen M |
| Citation(s) |
25330373 |
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| Submission date |
Oct 11, 2016 |
| Last update date |
May 27, 2018 |
| Contact name |
MAOSHAN CHEN |
| E-mail(s) |
maoshanchen@gmail.com
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| Organization name |
La Trobe University
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| Department |
Department of Biochemistry and Genetics
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| Street address |
La Trobe University, Bundoora
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| City |
Melbourne |
| State/province |
Victoria |
| ZIP/Postal code |
3086 |
| Country |
Australia |
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| Platforms (1) |
| GPL22548 |
Applied Biosystems Taqman Array Human MicroRNA A+B Cards Set v3.0 |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA347836 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE87839_fold-change.txt.gz |
7.9 Kb |
(ftp)(http) |
TXT |
| GSE87839_non-normalized.txt.gz |
9.4 Kb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
| Processed data are available on Series record |
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