Cell lines and cell culture

Human HCC cell lines Huh-7 (Japanese Cancer Research Resources Bank, catalog no. JCRB0403), PLC/PRF/5 [American Type Culture Collection (ATCC), catalog no. CRL-8024], HepG2 (ATCC, catalog no. HB-8065), Hep3B (ATCC, catalog no. HB-8064), and MHCC97-L (Beyotime, catalog no. C6586) (gifts from C. L. C. Wong in the University of Hong Kong), human breast cancer cell line MCF-7 (ATCC, catalog no. HTB-22), cervical cancer cell line HeLa (ATCC, catalog no. CRM-CCL-2), colon cancer cell line CT26 (ATCC, catalog no. CRL-2638), and non-small cell lung cancer cell line A549 (ATCC, catalog no. CRM-CCL-185) were cultured in Dulbecco's Modified Eagle Medium (DMEM; HyClone, Logan, UT) with 10% fetal bovine serum (FBS; HyClone) and 1% penicillin/streptomycin (HyClone). All cells were authenticated and maintained in a cell culture incubator under a 95% air and 5% CO₂ humidified environment at 37 °C and passaged every 3 to 4 days with 0.25% trypsin (HyClone). The cells used for the experiments involved in this study were no more than 20 passages.