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

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dc.contributorSistema FMUSP-HC: Faculdade de Medicina da Universidade de São Paulo (FMUSP) e Hospital das Clínicas da FMUSP
dc.contributor.authorCAMARGO, Juliana A.
dc.contributor.authorVIANA, Nayara I.
dc.contributor.authorPIMENTA, Ruan
dc.contributor.authorGUIMARAES, Vanessa R.
dc.contributor.authorSANTOS, Gabriel A. dos
dc.contributor.authorCANDIDO, Patricia
dc.contributor.authorGHAZARIAN, Vitoria
dc.contributor.authorROMAO, Poliana
dc.contributor.authorSILVA, Iran A.
dc.contributor.authorBIRBRAIR, Alexander
dc.contributor.authorSROUGI, Miguel
dc.contributor.authorNAHAS, William C.
dc.contributor.authorLEITE, Katia R.
dc.contributor.authorTRARBACH, Ericka B.
dc.contributor.authorREIS, Sabrina T.
dc.date.accessioned2023-12-15T18:41:43Z
dc.date.available2023-12-15T18:41:43Z
dc.date.issued2023
dc.description.abstractProstate 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.eng
dc.description.indexMEDLINE
dc.description.indexPubMed
dc.description.indexWoS
dc.description.indexScopus
dc.description.sponsorshipSao Paulo Research Foundation (FAPESP) [2018/19906-3, 2019/00156-7, 2019/19138-9, 2017/17362-3, 2022/02288-0]
dc.identifier.citationINTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v.24, n.19, article ID 14847, 15p, 2023
dc.identifier.doi10.3390/ijms241914847
dc.identifier.eissn1422-0067
dc.identifier.issn1661-6596
dc.identifier.urihttps://observatorio.fm.usp.br/handle/OPI/57299
dc.language.isoeng
dc.publisherMDPIeng
dc.relation.ispartofInternational Journal of Molecular Sciences
dc.rightsopenAccesseng
dc.rights.holderCopyright MDPIeng
dc.subjectCRISPR-Cas9eng
dc.subjectmiR-21eng
dc.subjectmatrix metalloproteinaseseng
dc.subjectmetastatic prostate cancereng
dc.subject.othermatrix metalloproteinaseseng
dc.subject.otherexpressioneng
dc.subject.otherinvasioneng
dc.subject.otherreckeng
dc.subject.wosBiochemistry & Molecular Biologyeng
dc.subject.wosChemistry, Multidisciplinaryeng
dc.titleThe Effect of Gene Editing by CRISPR-Cas9 of miR-21 and the Indirect Target MMP9 in Metastatic Prostate Cancereng
dc.typearticleeng
dc.type.categoryoriginal articleeng
dc.type.versionpublishedVersioneng
dspace.entity.typePublication
hcfmusp.affiliation.countryEstados Unidos
hcfmusp.affiliation.countryisous
hcfmusp.author.externalBIRBRAIR, Alexander:Univ Fed Minas Gerais, Dept Pathol, BR-30190002 Belo Horizonte, MG, Brazil; Univ Wisconsin Madison, Dept Dermatol, Madison, WI 53715 USA; Columbia Univ, Med Ctr, Dept Radiol, New York, NY 10032 USA
hcfmusp.citation.scopus1
hcfmusp.contributor.author-fmusphcJULIANA ALVES DE CAMARGO
hcfmusp.contributor.author-fmusphcNAYARA IZABEL VIANA MOURA
hcfmusp.contributor.author-fmusphcRUAN CESAR APARECIDO PIMENTA
hcfmusp.contributor.author-fmusphcVANESSA RIBEIRO GUIMARAES SCHREITER
hcfmusp.contributor.author-fmusphcGABRIEL ARANTES GALVAO DIAS DOS SANTOS
hcfmusp.contributor.author-fmusphcPATRICIA RODRIGUES CANDIDO
hcfmusp.contributor.author-fmusphcVITORIA GHAZARIAN NUNES
hcfmusp.contributor.author-fmusphcPOLIANA ROMAO DA SILVA
hcfmusp.contributor.author-fmusphcIRAN AMORIM DA SILVA
hcfmusp.contributor.author-fmusphcMIGUEL SROUGI
hcfmusp.contributor.author-fmusphcWILLIAM CARLOS NAHAS
hcfmusp.contributor.author-fmusphcKATIA RAMOS MOREIRA LEITE
hcfmusp.contributor.author-fmusphcERICKA BARBOSA TRARBACH
hcfmusp.contributor.author-fmusphcSABRINA THALITA DOS REIS FARIA
hcfmusp.description.articlenumber14847
hcfmusp.description.issue19
hcfmusp.description.volume24
hcfmusp.origemWOS
hcfmusp.origem.pubmed37834295
hcfmusp.origem.scopus2-s2.0-85174675570
hcfmusp.origem.wosWOS:001086026300001
hcfmusp.publisher.cityBASELeng
hcfmusp.publisher.countrySWITZERLANDeng
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hcfmusp.scopus.lastupdate2024-04-12
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