Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line.
<h4>Background</h4>Exosomes play a major role in cell-to-cell communication, targeting cells to transfer exosomal molecules including proteins, mRNAs, and microRNAs (miRNAs) by an endocytosis-like pathway. miRNAs are small noncoding RNA molecules on average 22 nucleotides in length that...
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2010
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oai:doaj.org-article:8d52301de72b403c969d56b6e6c290482021-11-18T07:03:32ZLet-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line.1932-620310.1371/journal.pone.0013247https://doaj.org/article/8d52301de72b403c969d56b6e6c290482010-10-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20949044/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Exosomes play a major role in cell-to-cell communication, targeting cells to transfer exosomal molecules including proteins, mRNAs, and microRNAs (miRNAs) by an endocytosis-like pathway. miRNAs are small noncoding RNA molecules on average 22 nucleotides in length that regulate numerous biological processes including cancer pathogenesis and mediate gene down-regulation by targeting mRNAs to induce RNA degradation and/or interfering with translation. Recent reports imply that miRNAs can be stably detected in circulating plasma and serum since miRNAs are packaged by exosomes to be protected from RNA degradation. Thus, profiling exosomal miRNAs are in need to clarify intercellular signaling and discover a novel disease marker as well.<h4>Methodology/principal findings</h4>Exosomes were isolated from cultured cancer cell lines and their quality was validated by analyses of transmission electron microscopy and western blotting. One of the cell lines tested, a metastatic gastric cancer cell line, AZ-P7a, showed the highest RNA yield in the released exosomes and distinctive shape in morphology. In addition, RNAs were isolated from cells and culture media, and profiles of these three miRNA fractions were obtained using microarray analysis. By comparing signal intensities of microarray data and the following validation using RT-PCR analysis, we found that let-7 miRNA family was abundant in both the intracellular and extracellular fractions from AZ-P7a cells, while low metastatic AZ-521, the parental cell line of AZ-P7a, as well as other cancer cell lines showed no such propensity.<h4>Conclusions/significance</h4>The enrichment of let-7 miRNA family in the extracellular fractions, particularly, in the exosomes from AZ-P7a cells may reflect their oncogenic characteristics including tumorigenesis and metastasis. Since let-7 miRNAs generally play a tumor-suppressive role as targeting oncogenes such as RAS and HMGA2, our results suggest that AZ-P7a cells release let-7 miRNAs via exosomes into the extracellular environment to maintain their oncogenesis.Keiichi OhshimaKanako InoueAkemi FujiwaraKeiichi HatakeyamaKaori KantoYuko WatanabeKoji MuramatsuYorikane FukudaShun-ichiro OguraKen YamaguchiTohru MochizukiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 10, p e13247 (2010) |
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DOAJ |
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EN |
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Medicine R Science Q |
| spellingShingle |
Medicine R Science Q Keiichi Ohshima Kanako Inoue Akemi Fujiwara Keiichi Hatakeyama Kaori Kanto Yuko Watanabe Koji Muramatsu Yorikane Fukuda Shun-ichiro Ogura Ken Yamaguchi Tohru Mochizuki Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line. |
| description |
<h4>Background</h4>Exosomes play a major role in cell-to-cell communication, targeting cells to transfer exosomal molecules including proteins, mRNAs, and microRNAs (miRNAs) by an endocytosis-like pathway. miRNAs are small noncoding RNA molecules on average 22 nucleotides in length that regulate numerous biological processes including cancer pathogenesis and mediate gene down-regulation by targeting mRNAs to induce RNA degradation and/or interfering with translation. Recent reports imply that miRNAs can be stably detected in circulating plasma and serum since miRNAs are packaged by exosomes to be protected from RNA degradation. Thus, profiling exosomal miRNAs are in need to clarify intercellular signaling and discover a novel disease marker as well.<h4>Methodology/principal findings</h4>Exosomes were isolated from cultured cancer cell lines and their quality was validated by analyses of transmission electron microscopy and western blotting. One of the cell lines tested, a metastatic gastric cancer cell line, AZ-P7a, showed the highest RNA yield in the released exosomes and distinctive shape in morphology. In addition, RNAs were isolated from cells and culture media, and profiles of these three miRNA fractions were obtained using microarray analysis. By comparing signal intensities of microarray data and the following validation using RT-PCR analysis, we found that let-7 miRNA family was abundant in both the intracellular and extracellular fractions from AZ-P7a cells, while low metastatic AZ-521, the parental cell line of AZ-P7a, as well as other cancer cell lines showed no such propensity.<h4>Conclusions/significance</h4>The enrichment of let-7 miRNA family in the extracellular fractions, particularly, in the exosomes from AZ-P7a cells may reflect their oncogenic characteristics including tumorigenesis and metastasis. Since let-7 miRNAs generally play a tumor-suppressive role as targeting oncogenes such as RAS and HMGA2, our results suggest that AZ-P7a cells release let-7 miRNAs via exosomes into the extracellular environment to maintain their oncogenesis. |
| format |
article |
| author |
Keiichi Ohshima Kanako Inoue Akemi Fujiwara Keiichi Hatakeyama Kaori Kanto Yuko Watanabe Koji Muramatsu Yorikane Fukuda Shun-ichiro Ogura Ken Yamaguchi Tohru Mochizuki |
| author_facet |
Keiichi Ohshima Kanako Inoue Akemi Fujiwara Keiichi Hatakeyama Kaori Kanto Yuko Watanabe Koji Muramatsu Yorikane Fukuda Shun-ichiro Ogura Ken Yamaguchi Tohru Mochizuki |
| author_sort |
Keiichi Ohshima |
| title |
Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line. |
| title_short |
Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line. |
| title_full |
Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line. |
| title_fullStr |
Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line. |
| title_full_unstemmed |
Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line. |
| title_sort |
let-7 microrna family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2010 |
| url |
https://doaj.org/article/8d52301de72b403c969d56b6e6c29048 |
| work_keys_str_mv |
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