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The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer
Prostate cancer (PCa) has a high prevalence and represents an important health problem, with an increased risk of metastasis. With the advance of CRISPR-Cas9 genome editing, new possibilities have been created for investigating PCa. The technique is effective in knockout oncogenes, reducing tumor re...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10573678/ https://www.ncbi.nlm.nih.gov/pubmed/37834295 http://dx.doi.org/10.3390/ijms241914847 |
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author | Camargo, Juliana A. Viana, Nayara I. Pimenta, Ruan Guimarães, Vanessa R. dos Santos, Gabriel A. Candido, Patrícia Ghazarian, Vitória Romão, Poliana Silva, Iran A. Birbrair, Alexander Srougi, Miguel Nahas, William C. Leite, Kátia R. Trarbach, Ericka B. Reis, Sabrina T. |
author_facet | Camargo, Juliana A. Viana, Nayara I. Pimenta, Ruan Guimarães, Vanessa R. dos Santos, Gabriel A. Candido, Patrícia Ghazarian, Vitória Romão, Poliana Silva, Iran A. Birbrair, Alexander Srougi, Miguel Nahas, William C. Leite, Kátia R. Trarbach, Ericka B. Reis, Sabrina T. |
author_sort | Camargo, Juliana A. |
collection | PubMed |
description | Prostate cancer (PCa) has a high prevalence and represents an important health problem, with an increased risk of metastasis. With the advance of CRISPR-Cas9 genome editing, new possibilities have been created for investigating PCa. The technique is effective in knockout oncogenes, reducing tumor resistance. MMP9 and miR-21 target genes are associated with PCa progression; therefore, we evaluated the MMP-9 and miR-21 targets in PCa using the CRISPR-Cas9 system. Single guide RNAs (sgRNAs) of MMP9 and miR-21 sequences were inserted into a PX-330 plasmid, and transfected in DU145 and PC-3 PCa cell lines. MMP9 and RECK expression was assessed by qPCR, WB, and IF. The miR-21 targets, integrins, BAX and mTOR, were evaluated by qPCR. Flow cytometry was performed with Annexin5, 7-AAD and Ki67 markers. Invasion assays were performed with Matrigel. The miR-21 CRISPR-Cas9-edited cells upregulated RECK, MARCKS, BTG2, and PDCD4. CDH1, ITGB3 and ITGB1 were increased in MMP9 and miR-21 CRISPR-Cas9-edited cells. Increased BAX and decreased mTOR were observed in MMP9 and miR-21 CRISPR-Cas9-edited cells. Reduced cell proliferation, increased apoptosis and low invasion in MMP9 and miR-21 edited cells was observed, compared to Scramble. CRISPR-Cas9-edited cells of miR-21 and MMP9 attenuate cell proliferation, invasion and stimulate apoptosis, impeding PCa evolution. |
format | Online Article Text |
id | pubmed-10573678 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-105736782023-10-14 The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer Camargo, Juliana A. Viana, Nayara I. Pimenta, Ruan Guimarães, Vanessa R. dos Santos, Gabriel A. Candido, Patrícia Ghazarian, Vitória Romão, Poliana Silva, Iran A. Birbrair, Alexander Srougi, Miguel Nahas, William C. Leite, Kátia R. Trarbach, Ericka B. Reis, Sabrina T. Int J Mol Sci Article Prostate cancer (PCa) has a high prevalence and represents an important health problem, with an increased risk of metastasis. With the advance of CRISPR-Cas9 genome editing, new possibilities have been created for investigating PCa. The technique is effective in knockout oncogenes, reducing tumor resistance. MMP9 and miR-21 target genes are associated with PCa progression; therefore, we evaluated the MMP-9 and miR-21 targets in PCa using the CRISPR-Cas9 system. Single guide RNAs (sgRNAs) of MMP9 and miR-21 sequences were inserted into a PX-330 plasmid, and transfected in DU145 and PC-3 PCa cell lines. MMP9 and RECK expression was assessed by qPCR, WB, and IF. The miR-21 targets, integrins, BAX and mTOR, were evaluated by qPCR. Flow cytometry was performed with Annexin5, 7-AAD and Ki67 markers. Invasion assays were performed with Matrigel. The miR-21 CRISPR-Cas9-edited cells upregulated RECK, MARCKS, BTG2, and PDCD4. CDH1, ITGB3 and ITGB1 were increased in MMP9 and miR-21 CRISPR-Cas9-edited cells. Increased BAX and decreased mTOR were observed in MMP9 and miR-21 CRISPR-Cas9-edited cells. Reduced cell proliferation, increased apoptosis and low invasion in MMP9 and miR-21 edited cells was observed, compared to Scramble. CRISPR-Cas9-edited cells of miR-21 and MMP9 attenuate cell proliferation, invasion and stimulate apoptosis, impeding PCa evolution. MDPI 2023-10-03 /pmc/articles/PMC10573678/ /pubmed/37834295 http://dx.doi.org/10.3390/ijms241914847 Text en © 2023 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 | Article Camargo, Juliana A. Viana, Nayara I. Pimenta, Ruan Guimarães, Vanessa R. dos Santos, Gabriel A. Candido, Patrícia Ghazarian, Vitória Romão, Poliana Silva, Iran A. Birbrair, Alexander Srougi, Miguel Nahas, William C. Leite, Kátia R. Trarbach, Ericka B. Reis, Sabrina T. The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer |
title | The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer |
title_full | The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer |
title_fullStr | The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer |
title_full_unstemmed | The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer |
title_short | The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer |
title_sort | effect of gene editing by crispr-cas9 of mir-21 and the indirect target mmp9 in metastatic prostate cancer |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10573678/ https://www.ncbi.nlm.nih.gov/pubmed/37834295 http://dx.doi.org/10.3390/ijms241914847 |
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