Interplay Between Estrogen and Replication Origins in Breast Cancer DNA Amplification.

摘要

Which is the molecular mechanism that leads to DNA amplification and oncogenes activation in breast cancer cells. This project aims to understand the role of estrogen in inducing re-replication, thus leading to DNA amplifications. I worked on the establishment of cancer cell line models carrying an engineered replication origin that will be tested for undergoing DNA amplification after estrogen treatment. I performed two biological replicates of a preliminary experiment done during Year-One, but I was unable to reproduce the exciting data previously obtained: Drug selection after estrogen exposure was not efficient in selecting cells with DNA amplification at the ectopic replication origin. Optimizing the working system to have multiple possibilities on the selection of cells with DNA amplification induced by estrogen receptor alpha, a new reporter construct have been built and a more efficient cell line model (U2OS/ER-alpha) have been characterized.