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...

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Bibliographic Details
Published in:International journal of molecular sciences 2023-10, Vol.24 (19), p.14847
Main Authors: 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
Format: Article
Language:English
Subjects:
Androgens
Apoptosis
CRISPR
Extracellular matrix
Gene expression
Genes
Genetic aspects
Genome editing
Genomes
Genomics
Integrins
Metastasis
MicroRNAs
Plasmids
Prostate cancer
Protein expression
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Summary: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.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms241914847