Abstract P2-16-07: Overexpression of the alpha subunit of the epithelial sodium channel in MDAMB231 breast cancer cells reduces cell migration and proliferation

Abstract Breast cancer is the most common cancer diagnosed in women worldwide and leads to 600,000+ deaths annually. The vast majority of breast cancer-related deaths are attributed to metastasis from the primary site in the breast to visceral organs or the brain. Therefore, understanding how these cells achieve metastasis is vital. Ion channels, such as the epithelial sodium channel (ENaC), are emerging as new targets for cancer research due to their role in regulating cell functions such as the process that allows cancer cells to undergo phenotypic changes necessary for metastasis. Ion channel dysregulation may be crucial in cancer development due to the ion channels’ potential role in controlling cancer cell characteristics such as migration and proliferation. There is limited research investigating the role ENaC may have in breast cancer. We have shown that more migratory, mesenchymal breast cancer cell lines have reduced mRNA and protein expression of ENaC compared to more epithelial breast cancer cell lines. Importantly, higher expression levels of the pore-forming alpha-ENaC subunit is correlated with an increased breast cancer survival in patients (n=1705) included in the SCAN-B data set. Bioinformatic analysis, using the SCAN-B data set, showed an increase in the expression of alpha-ENaC mRNA correlated with lower expression levels of migration and proliferation markers, suggesting ENaC has a role in breast cancer cell proliferation and migration. Therefore, this research examines the influence of stably overexpressing the pore-forming subunit of ENaC, alpha-ENaC, in breast cancer cells on their cell migration and proliferation ability. We hypothesised that increasing the expression of ENaC will restore the cells to a more epithelial state, thus reducing cell migration and proliferation. MDAMB231 breast cancer cells were engineered via a stable transfection to overexpress alpha-ENaC or to express an empty vector control. Several clones were isolated and expanded and using RTqPCR, alpha-ENaC mRNA overexpression was confirmed to consistently have 200-fold increased mRNA expression compared to control cells (n=9, p=0.0006). An increase in alpha-ENaC protein expression was also confirmed via western blotting. Using an EdU assay the alpha-ENaC overexpressing cells were shown to have a significant reduction in proliferation compared to the control cell line (n=3, p=0.0048). The alpha-ENaC overexpressing cells showed reduced migratory ability when examined in two established migration assays. In scratch wound assays, the alpha-ENaC overexpressing cells had a significant reduction in cell migration at 24 hours post scratch (n=4, p=0.0001). In Boyden chamber assays, the number of alpha-ENaC-overexpressing cells that migrated through the membrane was significantly reduced compared to the control cell line (n=4, p=0.02). Our results suggest that increased sodium entry into the cell through ENaC has a role in preventing the development of metastatic breast cancer cells, and highlights ENaC as a potential target for future breast cancer therapies. Citation Format: Sarah McQueen, Adam Ware, Heather Cunliffe, Fiona McDonald. Overexpression of the alpha subunit of the epithelial sodium channel in MDAMB231 breast cancer cells reduces cell migration and proliferation [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-16-07.