Hepatic Stellate Cell Coculture Enables Sorafenib Resistance in Huh7 Cells through HGF/c-Met/Akt and Jak2/Stat3 Pathways
Figure 2
Elevated HGF in LX2 coculture supernatant contributed to sorafenib resistance. (a) HGF increased in LX2 supernatants. HGF concentration in LX2 supernatants was determined by ELISA kits. HGF in LX2 supernatants increased more than 5 times than control medium to the concentration of almost 1300 pg/mL. (b) ETS-1 expression level in LX2 cocultured Huh7 was determined by real-time PCR. ETS-1 mRNA level increased about 3 times than control. (c) Administration of crizotinib reversed sorafenib resistance. 500 nM crizotinib was added to the coculture system for 48 h when administrating sorafenib. Cell viability was assessed by MTT assay. Crizotinib reversed coculture induced cell survival while it alone impaired no cell viability. (d) Crizotinib reversed inactivation of caspase cleavage by inhibiting p-Met, p-ERK, and p-Akt. Crizotinib inhibited phosphorylation of Met, ERK, and Akt while it did not affect Stat3 ( LX2 versus control; LX2 versus LX2 + crizotinib. sora: sorafenib).
