Establishment of a cell line with permanent silencing of PKCbI by shRNA and the impact on its phenotype

• Grant number: 15/20227-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: April 01, 2016
• End date: March 31, 2017
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Daniela Sanchez Basseres
• Grantee: Estela Maura Mesquita Carabette
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Protein kinases are essential and highly conserved in diverse organisms. Amongst the kinases we find protein kinase C (PKC) that phosphorylate the hydroxyl of serine and threonine residues. This family of kinases is composed of 10 different is enzymes sub-divided according to their activation requirements. PKCs are usually found in the cytoplasm when they are inactive and translocate to membranes upon activation. However, in recent years PKCs have been shown toll localize to the cellular nucleus, participating in diverse processes such as: regulation of DNA replication, RNA synthesis, chromatin structure, and control of gene expression. Our group observed that in breast cancer estrogen receptor positive, MCF-7 cells, PKCs bI and bII, are expressed in the nucleus. However, the role of these is enzymes in the nucleus of these cells and in breast cancer, is still not well understood. Thus, in the present project we intend to silence PKCb in MCF-7 cells, using short hairpin RNA (shRNA) and lentivirus, to establish a cell line that does not express PKCb and observe the effect of nuclear PKCb deletion in MCF-7 cells on proliferation and colony formation.