Modelling the risk factors of breast cancer in vitro

Abstract

Breast cancer accounts for around 25% of all newly-diagnosed cancers, and is one of the leading causes of cancer-related death in women due to a high rate of relapse and metastasis in patients undergoing remission. Epidemiological studies have identified obesity and alcohol consumption as major risk factors, which may be due to the inductions of partial epithelial to mesenchymal transition (p-EMT) and metabolic reprogramming caused by high levels of circulating leptin and ethanol. However, our understanding of the key signalling pathways involved in these inductions is, presently, somewhat limited. Additionally, caveats of traditional human cell culture media have led to the recent development of physiological media, which better simulate the microenvironmental conditions of cells in vivo. However, the impact of these media on the expression of stress-related genes remains somewhat explored. This project aims to study the expression of key EMT, metabolic, and pluripotency genes in response to traditional (DMEM/F-12) and physiological (Plasmax) media, as well as treatment with leptin and ethanol, in untransformed (non-cancerous) and transformed (cancerous) breast epithelial cells. Analysis of the expression of these genes does not clearly show whether or not Plasmax relieves stress on cells, but does suggest that it provides a more physiologically stable environment. Analysis also somewhat supports the roles of p-EMT and metabolic reprogramming in the breast cancer predisposition and progression facilitated by leptin and ethanol, and highlights some of the key players involved. Although these results are encouraging, a more complete dataset with sufficient biological replicates is required for robust conclusions to be drawn. Further experiments such as mammosphere formation assays, proliferation assays, and the construction of a pCAG-Fucci2a cell line (to investigate cell cycle changes) could further highlight the importance of physiological media, and elucidate the mechanisms underlying potential tumour-promoting signalling downstream of the two major breast cancer risk factors.

Date of Award	9 May 2023
Original language	English
Awarding Institution	University of Bristol
Supervisor	Abdelkader Essafi (Supervisor)