F12 medium

The SV-HUC-1, T24, BIU-87 and 5637 cell lines used in this study were purchased from the Chinese Academy of Science in Shanghai. SV-HUC-1 cell line were cultured F12 medium (Procell), T24, BIU-87 and 5637 cell lines were cultured RPMI-1640 medium (Procell) and supplemented with 10% FBS (Biological Industries) at 37°C in a 5% CO₂ humidified atmosphere.

Human NSCLC cells A549 and human normal lung epithelial cells BEAS-2B were provided by the Tongji Hospital, Affiliated with Tongji Medical College of Huazhong University of science and technology. We purchased human NSCLC cells H1944 (cat#CL-0632) from Procell. A Mycoplasma contamination test was carried out, which was negative. A549 cells were routinely cultured in F12 medium (Procell, cat#PM150312), BEAS-2B cells were routinely cultured in DMEM with high glucose (Gibco, cat#11965092), and the H1944 cells were routinely cultured in RPMI-1640 medium (Procell, cat#PM150110). All the media were supplemented with 1% penicillin and streptomycin solution (Beyotime, cat#ST488) and 10% fetal bovine serum (Gibco, cat#A3160902). The cells were incubated at 37 °C under 5% CO₂. Nutlin-3 (Glpbio, cat#GC16051) and Simvastatin (SIM, Selleck, cat#S1792) powder were dissolved in DMSO to prepare a stock solution. The working solution was diluted to 10 μM in the medium. DL-Mevalonolactone (DL-MVA) (Molnova, cat#M15570) and Methoxyestradiol (2-MeOE2, Selleck, cat#S1233) solutions were dissolved in a medium and respectively diluted to 100 μM and 10 μM.