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

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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.

• Tipo de documento: Artigo Científico
• Fonte: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES; v. 24, n. 19, p. 15-pg., 2023-10-01.

Resumo

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)

• Processo FAPESP: 18/19906-3 - Edição genômica com CRISPR/Cas9 para avaliação do papel da MMP9 e seu regulador MIR21 no Câncer de Próstata
• Beneficiário: Juliana Alves de Camargo
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 19/19138-9 - Estudo do efeito do miR-137 sobre linhagem celular de Câncer de Próstata resistente a castração em ambiente hipercolesterolêmico
• Beneficiário: Vitória Ghazarian Nunes
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 17/17362-3 - Edição genômica com CRISPR/Cas9 para avaliação do papel da MMP9 e de seu regulador MIR21 no câncer de próstata
• Beneficiário: Sabrina Thalita dos Reis Faria
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 19/00156-7 - Estudo in vivo de coativadores e corepressores do receptor de andrógeno no Câncer de Próstata
• Beneficiário: Ruan César Aparecido Pimenta
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 22/02288-0 - Avaliação do efeito do silenciamento da MMP9 pelo sistema CRISPR-Cas9 concomitante ao uso do BCG e anti-PDL1 no tratamento do Câncer de Bexiga: estudo in vivo
• Beneficiário: Sabrina Thalita dos Reis Faria
• Modalidade de apoio: Auxílio à Pesquisa - Regular