Adipocytes induced distinct gene expression profiling of triple negative breast cancer cells

We investigated how adiposity reprograms the metabolism and progression of triple-negative breast cancer (TNBC) cells compared to non-TNBC cells. MDA-MB-468 (TNBC) and BT474 (non-TNBC) cells were co-cultured with mature adipocytes, followed by RNA-seq analysis to assess the metabolic impact of adipocyte on TNBC cells. Overall design: MDA-MB-468 and BT-474 cells were cultured with DMEM/F12 complete medium containing 10%FBS. The medium used for human adipocytes was a high-glucose DMEM medium containing 10% FBS.using 10% FBS, 0.5 mM IBMX, 1 µM dexamethasone, and 10 µg/mL insulin for 3 days. Differentiation was further promoted with DMEM containing 10% FBS and 10 µg/mL insulin for 3 days, followed by maintenance in DMEM with 10% FBS for an additional 4 days until mature adipocytes, rich in lipid droplets, were fully formed. Subsenquently, breast cancer cells were cocultured with mature adipocytes via transwell coculture system for 72 hours. Total RNA of breast cancer cells with the presence or absence of adipocytes were conducted for RNA sequencing (RNA-seq) .