Development of Sorafenib-Resistant Hep3B Cells

Resistant Hep3B cells were developed in clinically relevant models [116 (link)], as previously reported [117 (link)]. In T-75 flasks, Hep3B cells were seeded overnight and incubated at 37 °C. The cells were then treated with sorafenib at the 10% inhibitory concentration (IC10) (0.4 µM) of sorafenib (Biovision, Milpitas, CA, USA #BAY 43-9006) to mimic the clinically consumed low dose of chemotherapeutic drugs by a cancer patient who is then exposed to escalating doses over time [116 (link)]. To develop resistance, the survived cells were transplanted to a new flask and then treated with sorafenib at escalating concentrations over six months. Then, sorafenib at concentration IC10 (0.4 µM) was persistently retained in the culture media to ensure and maintain resistance. MTT cell viability assay was performed each month to validate the resistance behavior of the cells.