CRISPR/Cas9 – Mediated MMP-9 Silencing Inhibits Bladder Cancer T24 Cell Invasion and Migration In Vitro

• DOI: 10.2139/ssrn.4412721
• Publication Date: 2023-04-14T16:46:40Z
• Authors: Fabio Pescarmona...; Juliana Alves Ca...; Nayara Izabel Vi...; Ruan Cesar Apare...; Vanessa Ribeiro ...; Katia Ramos More...; William Carlos N...; Sabrina Thalita ...

Background: Bladder Cancer (BCa) is the tenth most common type of cancer, presenting significant morbidity, mortality, and healthcare costs, especially in more advanced cases. New biomarkers disease mechanisms oncology have been studied, among them matrix metalloproteinases (MMPs) their role cancer progression metastasis. Therefore, we analyzed MMP-9 gene expression downregulation effects by CRISPR-Cas9 genome editing technique T24-luc bladder urothelial carcinoma cell lines.Materials methods: Insertion guide RNA sequences (sgRNA 1 2) into pX-330 plasmids targeting was performed. The resultant were transfected BCa cells, protein using RT-qPCR Western blotting. To analyze impact silencing, proliferation apoptosis assays performed flow cytometry, colony formation assay, migration assay wound healing test, Matrigel™ invasion assay.Results: After transfecting T24 line, a reduction observed sgRNA2, for both sgRNAs. Reduced capacity, increased cellular reduced vitro capacity edited cells observed. A cells' also observed.Conclusions: silencing line inhibited proliferation, migration, invasion, apoptosis. These results support tumorigenesis processes suggest as potential therapeutic target.