Sistema FMUSP-HC: Faculdade de Medicina da Universidade de São Paulo (FMUSP) e Hospital das Clínicas da FMUSPCAMARGO, Juliana A.VIANA, Nayara I.PIMENTA, RuanGUIMARAES, Vanessa R.SANTOS, Gabriel A. dosCANDIDO, PatriciaGHAZARIAN, VitoriaROMAO, PolianaSILVA, Iran A.BIRBRAIR, AlexanderSROUGI, MiguelNAHAS, William C.LEITE, Katia R.TRARBACH, Ericka B.REIS, Sabrina T.2023-12-152023-12-152023INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v.24, n.19, article ID 14847, 15p, 20231661-6596https://observatorio.fm.usp.br/handle/OPI/57299Prostate 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.engopenAccessCRISPR-Cas9miR-21matrix metalloproteinasesmetastatic prostate cancermatrix metalloproteinasesexpressioninvasionreckarticleCopyright MDPI10.3390/ijms241914847Biochemistry & Molecular BiologyChemistry, Multidisciplinary1422-0067