USE OF SHRNA SCREEN TO IDENTIFIY UBIQUITIN RELATED GENES INVOLVED IN THE REGULATION OF TUMOR-CELL MIGRATION AND INVASION

JULIANA H. ENRIQUE´ STEINBERG; MARIO ROSSI; EZEQUIEL CALVO-ROITBERG ; FABIANA ALEJANDRA ROSSI; JOAQUIN M ESPINOSA

Congreso; Reunión Conjunta de Sociedades de Biociencias 2017; 2017

The development of metastasis foci in patients suffering from cancer represents a significant reduction in their survival and life quality. The Ubiquitin-Proteasome System (UPS) plays a fundamental role in the maintenance of protein homeostasis both in normal and stressed conditions, thus regulating almost every single cellular process. Since alterations in the ubiquitination cascade have been shown to be associated with malignant transformation, invasive potential of cells and metastasis, we sought to investigate the role of the UPS in the regulation of tumor-cell migration and invasion. To this end we performed a genetic screen using a shRNA library against UPS associated genes, and Boyden chambers. After the selection process, we characterized the non-migrating cell population and determined the relative abundance of each shRNA by Illumina NGS sequencing. We obtained a list of 30 candidate genes, half of which had already been associated with regulation of migration/invasion/tumorigenesis processes or metastasis. Among the candidates, we focused on a specific DUB and demonstrated that its silencing reduces the migratory/invasive potential of different tumor-cell lines using Boyden chamber (p