Use of shRNA screen to identify ubiquitin related genes involved in the regulation of tumor cell migration and invasion

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

Simposio; Frontiers in Bioscience III; 2018

Despite formidable advance in the prevention, early detection and treatment of a great number of cancers, the development of metastasis foci in patients suffering from this disease still represents a significant reduction in their survival and life quality. Although metastatic cells have partially been characterized, there is yet not an effective treatment for this pathological condition. The Ubiquitylation System plays a fundamental role in the maintenance of protein homeostasis both in normal and stressed conditions, in almost every single cellular process. Since alterations in the ubiquitylation cascade have been shown to be associated with malignant transformation, invasive potential of cells and metastasis, we sought to investigate its role in the regulation of tumor-cell invasion and migration. We performed a genetic screen using a shRNA library against Ubiquitin-related genes, and Boyden chambers to analyze the migrating/invasive potential of breast cancer derived cells, infected with this library. After the selection process, the non-migrating populations of cells were characterized and the shRNA present in them were determined. We validated USP19 as a novel regulator of cell migration and invasion both in vitro and in vivo