(662.2) Mass Spectrometry based Proteomics to Investigate and Characterize the Human Jumping Translocation Breakpoint (hJTB) Protein using Cancer Cell Lines
Monday, April 4, 2022
12:30 PM – 1:45 PM
Location: Exhibit/Poster Hall A-B - Pennsylvania Convention Center
Presenting Author Clarkson University Potsdam, New York
Human JTB (hJTB) is a gene located on the human chromosome 1 at q21 which is involved in the unbalanced translocation in various types of cancer. JTB protein is ubiquitously present in normal cells and is found to be overexpressed in various types of cancer including prostate and breast cancer. Hence this protein could be a biomarker for tumor malignancies and a potential target for their treatment. However, the biological function and the pathway through which this protein causes increased cell growth and proliferation is not entirely clear. Investigation and comparison of the proteomes of cells with upregulated and downregulated JTB can be a good approach to understand the function of the protein and also its contribution to tumorigenesis. In this study, MCF7 breast cancer cell lines were transfected with the sense orientation of the JTB cDNA in HA, His and FLAG tagged CMV expression vector as well as with shRNA plasmids. Proteins extracted from transient and stable transfected cells were separated using SDS-PAGE. The expression of JTB was confirmed by western blotting technique. In gel digested peptides were analyzed by a Nano Acquity UPLC coupled with QTOF Xevo G2 Mass Spectrometer. Data processing was done using Mascot 2.4 server and Scaffold 4.1 software. We found several proteins that were dysregulated. Furthermore, we will do Immunoprecipitation to look at JTB protein interacting partners. These studies could help us elucidate the mechanism through which JTB induces cell proliferation and test the JTB protein as a potential drug target for malignancies with overexpression of the protein.