Genetic Dissection of PTEN Signaling Mechanisms in Prostate Cancer

摘要

The goal of my traineeship is to employ genetic approaches to investigate PTEN signaling in prostate cancer. My first task was to construct expression vectors for a series of tumor-derived PTEN mutations and test these for the ability to perform a diverse set of PTEN functions. This mutant analysis is 75% complete and will be submitted for publication this year. My second task was to set-up and perform a large-scale RNAi screen to identify novel components involved in PTEN signaling. Setting-up this screening system has proven to be very difficult. After trying many approaches it appears that we have finally identified a tractable screening system. Initially, we tried to set-up to do a luciferase reporter screen, but the dynamic range was not suitable for screening. We switched to doing a suppressor screen using inducible PTEN. My second year of funding will primarily focus on performing this screen. While setting up the PTEN RNAi screen we found that a TRAIL luciferase reporter can be used to monitor endogenous Pi3K pathway activity from the insulin receptor to the transcriptional outputs; this work now comprises a full manuscript which will be submitted for publication.