Cargando…
Secretion of Mutant DNA and mRNA by the Exosomes of Breast Cancer Cells
Exosomes are the small vesicles that are secreted by different types of normal and tumour cells and can incorporate and transfer their cargo to the recipient cells. The main goal of the present work was to study the tumour exosomes’ ability to accumulate the parent mutant DNA or RNA transcripts with...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8123307/ https://www.ncbi.nlm.nih.gov/pubmed/33922925 http://dx.doi.org/10.3390/molecules26092499 |
_version_ | 1783692865383170048 |
---|---|
author | Andreeva, Olga E. Shchegolev, Yuri Y. Scherbakov, Alexander M. Mikhaevich, Ekaterina I. Sorokin, Danila V. Gudkova, Margarita V. Bure, Irina V. Kuznetsova, Ekaterina B. Mikhaylenko, Dmitry S. Nemtsova, Marina V. Bagrov, Dmitry V. Krasil’nikov, Mikhail A. |
author_facet | Andreeva, Olga E. Shchegolev, Yuri Y. Scherbakov, Alexander M. Mikhaevich, Ekaterina I. Sorokin, Danila V. Gudkova, Margarita V. Bure, Irina V. Kuznetsova, Ekaterina B. Mikhaylenko, Dmitry S. Nemtsova, Marina V. Bagrov, Dmitry V. Krasil’nikov, Mikhail A. |
author_sort | Andreeva, Olga E. |
collection | PubMed |
description | Exosomes are the small vesicles that are secreted by different types of normal and tumour cells and can incorporate and transfer their cargo to the recipient cells. The main goal of the present work was to study the tumour exosomes’ ability to accumulate the parent mutant DNA or RNA transcripts with their following transfer to the surrounding cells. The experiments were performed on the MCF7 breast cancer cells that are characterized by the unique coding mutation in the PIK3CA gene. Using two independent methods, Sanger sequencing and allele-specific real-time PCR, we revealed the presence of the fragments of the mutant DNA and RNA transcripts in the exosomes secreted by the MCF7 cells. Furthermore, we demonstrated the MCF7 exosomes’ ability to incorporate into the heterologous MDA-MB-231 breast cancer cells supporting the possible transferring of the exosomal cargo into the recipient cells. Sanger sequencing of the DNA from MDA-MB-231 cells (originally bearing a wild type of PIK3CA) treated with MCF7 exosomes showed no detectable amount of mutant DNA or RNA; however, using allele-specific real-time PCR, we revealed a minor signal from amplification of a mutant allele, showing a slight increase of mutant DNA in the exosome-treated MDA-MB-231 cells. The results demonstrate the exosome-mediated secretion of the fragments of mutant DNA and mRNA by the cancer cells and the exosomes’ ability to transfer their cargo into the heterologous cells. |
format | Online Article Text |
id | pubmed-8123307 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-81233072021-05-16 Secretion of Mutant DNA and mRNA by the Exosomes of Breast Cancer Cells Andreeva, Olga E. Shchegolev, Yuri Y. Scherbakov, Alexander M. Mikhaevich, Ekaterina I. Sorokin, Danila V. Gudkova, Margarita V. Bure, Irina V. Kuznetsova, Ekaterina B. Mikhaylenko, Dmitry S. Nemtsova, Marina V. Bagrov, Dmitry V. Krasil’nikov, Mikhail A. Molecules Communication Exosomes are the small vesicles that are secreted by different types of normal and tumour cells and can incorporate and transfer their cargo to the recipient cells. The main goal of the present work was to study the tumour exosomes’ ability to accumulate the parent mutant DNA or RNA transcripts with their following transfer to the surrounding cells. The experiments were performed on the MCF7 breast cancer cells that are characterized by the unique coding mutation in the PIK3CA gene. Using two independent methods, Sanger sequencing and allele-specific real-time PCR, we revealed the presence of the fragments of the mutant DNA and RNA transcripts in the exosomes secreted by the MCF7 cells. Furthermore, we demonstrated the MCF7 exosomes’ ability to incorporate into the heterologous MDA-MB-231 breast cancer cells supporting the possible transferring of the exosomal cargo into the recipient cells. Sanger sequencing of the DNA from MDA-MB-231 cells (originally bearing a wild type of PIK3CA) treated with MCF7 exosomes showed no detectable amount of mutant DNA or RNA; however, using allele-specific real-time PCR, we revealed a minor signal from amplification of a mutant allele, showing a slight increase of mutant DNA in the exosome-treated MDA-MB-231 cells. The results demonstrate the exosome-mediated secretion of the fragments of mutant DNA and mRNA by the cancer cells and the exosomes’ ability to transfer their cargo into the heterologous cells. MDPI 2021-04-25 /pmc/articles/PMC8123307/ /pubmed/33922925 http://dx.doi.org/10.3390/molecules26092499 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Communication Andreeva, Olga E. Shchegolev, Yuri Y. Scherbakov, Alexander M. Mikhaevich, Ekaterina I. Sorokin, Danila V. Gudkova, Margarita V. Bure, Irina V. Kuznetsova, Ekaterina B. Mikhaylenko, Dmitry S. Nemtsova, Marina V. Bagrov, Dmitry V. Krasil’nikov, Mikhail A. Secretion of Mutant DNA and mRNA by the Exosomes of Breast Cancer Cells |
title | Secretion of Mutant DNA and mRNA by the Exosomes of Breast Cancer Cells |
title_full | Secretion of Mutant DNA and mRNA by the Exosomes of Breast Cancer Cells |
title_fullStr | Secretion of Mutant DNA and mRNA by the Exosomes of Breast Cancer Cells |
title_full_unstemmed | Secretion of Mutant DNA and mRNA by the Exosomes of Breast Cancer Cells |
title_short | Secretion of Mutant DNA and mRNA by the Exosomes of Breast Cancer Cells |
title_sort | secretion of mutant dna and mrna by the exosomes of breast cancer cells |
topic | Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8123307/ https://www.ncbi.nlm.nih.gov/pubmed/33922925 http://dx.doi.org/10.3390/molecules26092499 |
work_keys_str_mv | AT andreevaolgae secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT shchegolevyuriy secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT scherbakovalexanderm secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT mikhaevichekaterinai secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT sorokindanilav secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT gudkovamargaritav secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT bureirinav secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT kuznetsovaekaterinab secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT mikhaylenkodmitrys secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT nemtsovamarinav secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT bagrovdmitryv secretionofmutantdnaandmrnabytheexosomesofbreastcancercells AT krasilnikovmikhaila secretionofmutantdnaandmrnabytheexosomesofbreastcancercells |