The Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancer

Author(s):Show less -

Camargo, Juliana A. ; Viana, Nayara I. ; Pimenta, Ruan ; Guimaraes, Vanessa R. ; dos Santos, Gabriel A. ; Candido, Patricia ; Ghazarian, Vitoria ; Romao, Poliana ; Silva, Iran A. ; Birbrair, Alexander ; Srougi, Miguel ; Nahas, William C. ; Leite, Katia R. ; Trarbach, Ericka B. ; Reis, Sabrina T.

• Document type: Journal article
• Source: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES; v. 24, n. 19, p. 15-pg., 2023-10-01.

Abstract

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. (AU)

• FAPESP's process: 18/19906-3 - Genomic edition with CRISPR/Cas9 to evaluate the role of MMP9 and the regulator miR21 in Prostate Cancer
• Grantee: Juliana Alves de Camargo
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 19/19138-9 - Study of the effect of miR-137 on cell line of castration resistant prostate cancer in a hypercholesterolemic environment
• Grantee: Vitória Ghazarian Nunes
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 17/17362-3 - Genomic edition with CRISPR / Cas9 to evaluate the role of MMP9 and it regulator MIR21 in prostate cancer
• Grantee: Sabrina Thalita dos Reis Faria
• Support Opportunities: Regular Research Grants
• FAPESP's process: 19/00156-7 - In vivo study of co-activators and co-repressors of androgen receptor in Prostate Câncer
• Grantee: Ruan César Aparecido Pimenta
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 22/02288-0 - Evaluation of the effect of MMP9 silencing by CRISPR-Cas9 system concomitant to the use of BCG and anti-PDL1 in the treatment of Bladder Cancer: in vivo study
• Grantee: Sabrina Thalita dos Reis Faria
• Support Opportunities: Regular Research Grants