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Analysis of miR-221/222 blockade in vitro model of renal cell carcinoma clear cell non-metastatic and metastatic

Grant number: 23/02352-3
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): June 01, 2023
Effective date (End): May 31, 2024
Field of knowledge:Health Sciences - Medicine - Surgery
Principal Investigator:Sabrina Thalita dos Reis Faria
Grantee:Giovanna Tamarindo da Silveira
Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Renal cell carcinoma (RCC) is a malignant neoplasm that originates in the lining of the proximal convoluted tubule of the nephrons. The disease is responsible for 2% of all cancer diagnoses and deaths in the world and 85% of all kidney tumors, being classified as the seventh most common form of cancer in the developed world. Clear cell renal cell carcinoma (ccRCC) is the most common subtype, accounting for 75% of renal carcinomas. ccRCC is associated with autosomal dominant Von Hippel Lindau syndrome, where the VHL tumor suppressor gene is inactivated. In recent years, groups of microRNAs (miRNA) have been found to have aberrant expression in neoplastic tissues including ccRCC, attracting increasing attention due to their associations with CRCC prognosis and the potential to become therapeutic targets. miR-221/222 can be up or down-regulated in several types of cancer, and in ccRCC, a study showed that in addition to the aberrant expression of miR 221 and 222 in tissues of patients with ccRCC, ectopic overexpression of miRNA 221 and 222 222 results in the reduction of the p27 gene (Kip-1), an inhibitor of cyclin-dependent kinases, with a crucial role in regulating the cell cycle. Here in our study, we will evaluate these microRNAs´ inhibition effect in metastatic and non-metastatic ccRCC cell lines. The cells to be used belong to two different strains: 786-O and Caki-1 which will then be transfected with anti-miR-221 and 222. After transfection we will extract the miRNA and evaluate the expression of miR221 and 222 by qPCR for analysis of transfection efficiency. To evaluate the effect of blocking these microRNAs, we will perform invasion, migration and colony formation assays.

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