Transcriptomic Profiling of Senescent versus Non-senescent Cancer-Associated Fibroblasts in Colorectal Cancer

Cancer-associated fibroblasts (CAFs) are a major component of the tumor microenvironment and play a critical role in cancer progression. Cellular senescence is a state of irreversible cell-cycle arrest that can paradoxically influence tumorigenesis. This study aims to elucidate the transcriptomic characteristics between senescent and non-senescent CAFs and investigates the role of senescent CAFs (sCAFs) in colorectal cancer (CRC) chemoresistance and the underlying mechanisms. Primary CAFs were isolated from in situ tumors of the MC38 murine colorectal cancer model. The isolation procedure involved enzymatic digestion of 1 mm3 tissue fragments with 1 mg/ml type IV collagenase at 37 C for 2 hours. The resulting cell suspension was filtered and cultured in DMEM high glucose medium with 10% fetal bovine serum. Non-adherent cells were removed after 72 hours of incubation. For senescence induction, cultured CAFs at 80% confluence were treated with bleomycin (50 ug/ml) for 24 hours. Following drug exposure, cells were washed and maintained in drug-free medium for 5 days to allow the senescence phenotype to fully develop. Non-treated, proliferating CAFs from the same passages were used as controls. We performed bulk RNA sequencing on four biological replicates of senescent CAFs (sCAF) and four biological replicates of non-senescent control CAFs (nsCAF) to identify gene expression signatures and pathways associated with CAF senescence. This dataset provides a valuable resource for understanding the functional roles of senescent CAFs in the colorectal cancer microenvironment and for identifying potential therapeutic targets.