Bulk RNA-seq of HFF cell senescence model induced by oncogene activation

To investiage the mechanism of cellular senescence, we established a model of oncogene RAS-induced cellular senescence in human foreskin fibroblasts (HFF cells). We transduced an estrogen receptor fused to the H-RASG12V protein (ER:RAS) in HFF, which can be induced by 4-hydroxytamoxifen (4-OHT). Lentiviral packaging of the oncogene-inducible expression vector (pLNC ER:RAS (G12V)) was conducted using HEK293T cells. The collected viral supernatant was then introduced into younger (~P18) HFF cells. Following infection, resistant clones were selected using 200 μg/ml of neomycin at 48-72 hours post-infection. After five days, stable transgenic cell lines were obtained. To activate the expression of H-RAS (G12V), 100 nM of 4-hydroxytamoxifen (4-OHT) was added, and cells exhibiting oncogene-induced senescence (OIS) were harvested one week thereafter.