Proteomic analysis of exosomes secreted during the epithelial-mesenchymal transition and potential biomarkers of mesenchymal high-grade serous ovarian carcinoma
Abstract Background The epithelial-mesenchymal transition (EMT) promotes cell signaling and morphology alterations, contributing to cancer progression. Exosomes, extracellular vesicles containing proteins involved in cell-cell communication, have emerged as a potential source of biomarkers for sever...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-11-01
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| Series: | Journal of Ovarian Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13048-023-01304-0 |
| _version_ | 1797414216923086848 |
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| author | Germano Aguiar Ferreira Carolina Hassibe Thomé Clarice Izumi Mariana Lopes Grassi Guilherme Pauperio Lanfredi Marcus Smolka Vitor Marcel Faça Francisco José Candido dos Reis |
| author_facet | Germano Aguiar Ferreira Carolina Hassibe Thomé Clarice Izumi Mariana Lopes Grassi Guilherme Pauperio Lanfredi Marcus Smolka Vitor Marcel Faça Francisco José Candido dos Reis |
| author_sort | Germano Aguiar Ferreira |
| collection | DOAJ |
| description | Abstract Background The epithelial-mesenchymal transition (EMT) promotes cell signaling and morphology alterations, contributing to cancer progression. Exosomes, extracellular vesicles containing proteins involved in cell-cell communication, have emerged as a potential source of biomarkers for several diseases. Methods Our aim was to assess the proteome content of exosomes secreted after EMT-induction to identify potential biomarkers for ovarian cancer classification. EMT was induced in the ovarian cancer cell line CAOV3 by treating it with EGF (10 ng/mL) for 96 h following 24 h of serum deprivation. Subsequently, exosomes were isolated from the supernatant using selective centrifugation after decellularization, and their characteristics were determined. The proteins present in the exosomes were extracted, identified, and quantified using Label-Free-Quantification (LFQ) via Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). To identify potential biomarkers, the obtained proteomic data was integrated with the TGGA database for mRNA expression using principal component analysis and a conditional inference tree. Results The exosomes derived from CAOV3 cells exhibited similar diameter and morphology, measuring approximately 150 nm, regardless of whether they were subjected to EMT stimulation or not. The proteomic analysis of proteins from CAOV3-derived exosomes revealed significant differential regulation of 157 proteins, with 100 showing upregulation and 57 downregulation upon EMT induction. Further comparison of the upregulated proteins with the TCGA transcriptomic data identified PLAU, LAMB1, COL6A1, and TGFB1 as potential biomarkers of the mesenchymal HGSOC subtype. Conclusions The induction of EMT, the isolation of exosomes, and the subsequent proteomic analysis highlight potential biomarkers for an aggressive ovarian cancer subtype. Further investigation into the role of these proteins is warranted to enhance our understanding of ovarian cancer outcomes. |
| first_indexed | 2024-03-09T05:29:53Z |
| format | Article |
| id | doaj.art-01d6e5bc76e64191a89bf145aa6cec0d |
| institution | Directory Open Access Journal |
| issn | 1757-2215 |
| language | English |
| last_indexed | 2024-03-09T05:29:53Z |
| publishDate | 2023-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Ovarian Research |
| spelling | doaj.art-01d6e5bc76e64191a89bf145aa6cec0d2023-12-03T12:33:02ZengBMCJournal of Ovarian Research1757-22152023-11-0116111310.1186/s13048-023-01304-0Proteomic analysis of exosomes secreted during the epithelial-mesenchymal transition and potential biomarkers of mesenchymal high-grade serous ovarian carcinomaGermano Aguiar Ferreira0Carolina Hassibe Thomé1Clarice Izumi2Mariana Lopes Grassi3Guilherme Pauperio Lanfredi4Marcus Smolka5Vitor Marcel Faça6Francisco José Candido dos Reis7Department of Gynecology and Obstetrics, Ribeirão Preto Medical SchoolDepartment of Gynecology and Obstetrics, Ribeirão Preto Medical SchoolDepartment of Cellular and Molecular Biology and Pathogenic Bioagents, Ribeirão Preto Medical School, University of São PauloDepartment of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São PauloDepartment of Biochemistry and Immunology, Ribeirão Preto Medical School, University of São PauloWeill Institute for Cell and Molecular Biology, Cornell UniversityRegional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, and Center for Cell Based Therapy, University of São PauloDepartment of Gynecology and Obstetrics, Ribeirão Preto Medical SchoolAbstract Background The epithelial-mesenchymal transition (EMT) promotes cell signaling and morphology alterations, contributing to cancer progression. Exosomes, extracellular vesicles containing proteins involved in cell-cell communication, have emerged as a potential source of biomarkers for several diseases. Methods Our aim was to assess the proteome content of exosomes secreted after EMT-induction to identify potential biomarkers for ovarian cancer classification. EMT was induced in the ovarian cancer cell line CAOV3 by treating it with EGF (10 ng/mL) for 96 h following 24 h of serum deprivation. Subsequently, exosomes were isolated from the supernatant using selective centrifugation after decellularization, and their characteristics were determined. The proteins present in the exosomes were extracted, identified, and quantified using Label-Free-Quantification (LFQ) via Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). To identify potential biomarkers, the obtained proteomic data was integrated with the TGGA database for mRNA expression using principal component analysis and a conditional inference tree. Results The exosomes derived from CAOV3 cells exhibited similar diameter and morphology, measuring approximately 150 nm, regardless of whether they were subjected to EMT stimulation or not. The proteomic analysis of proteins from CAOV3-derived exosomes revealed significant differential regulation of 157 proteins, with 100 showing upregulation and 57 downregulation upon EMT induction. Further comparison of the upregulated proteins with the TCGA transcriptomic data identified PLAU, LAMB1, COL6A1, and TGFB1 as potential biomarkers of the mesenchymal HGSOC subtype. Conclusions The induction of EMT, the isolation of exosomes, and the subsequent proteomic analysis highlight potential biomarkers for an aggressive ovarian cancer subtype. Further investigation into the role of these proteins is warranted to enhance our understanding of ovarian cancer outcomes.https://doi.org/10.1186/s13048-023-01304-0Ovarian cancerEpithelial-mesenchymal transitionExtracellular vesiclesExosomesSecretome |
| spellingShingle | Germano Aguiar Ferreira Carolina Hassibe Thomé Clarice Izumi Mariana Lopes Grassi Guilherme Pauperio Lanfredi Marcus Smolka Vitor Marcel Faça Francisco José Candido dos Reis Proteomic analysis of exosomes secreted during the epithelial-mesenchymal transition and potential biomarkers of mesenchymal high-grade serous ovarian carcinoma Journal of Ovarian Research Ovarian cancer Epithelial-mesenchymal transition Extracellular vesicles Exosomes Secretome |
| title | Proteomic analysis of exosomes secreted during the epithelial-mesenchymal transition and potential biomarkers of mesenchymal high-grade serous ovarian carcinoma |
| title_full | Proteomic analysis of exosomes secreted during the epithelial-mesenchymal transition and potential biomarkers of mesenchymal high-grade serous ovarian carcinoma |
| title_fullStr | Proteomic analysis of exosomes secreted during the epithelial-mesenchymal transition and potential biomarkers of mesenchymal high-grade serous ovarian carcinoma |
| title_full_unstemmed | Proteomic analysis of exosomes secreted during the epithelial-mesenchymal transition and potential biomarkers of mesenchymal high-grade serous ovarian carcinoma |
| title_short | Proteomic analysis of exosomes secreted during the epithelial-mesenchymal transition and potential biomarkers of mesenchymal high-grade serous ovarian carcinoma |
| title_sort | proteomic analysis of exosomes secreted during the epithelial mesenchymal transition and potential biomarkers of mesenchymal high grade serous ovarian carcinoma |
| topic | Ovarian cancer Epithelial-mesenchymal transition Extracellular vesicles Exosomes Secretome |
| url | https://doi.org/10.1186/s13048-023-01304-0 |
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