Effects of isorhynchophylline on proliferation,migration,and 5-FU resistance of liver cancer cells by regulating EGFR/FAK signaling pathway
Objective To investigate the effects of isorhynchophylline(isorhy)on the proliferation,migration,and 5-FU resistance of liver cancer cells by regulating the epidermal growth factor receptor(EGFR)/focal adhesion kinase(FAK)signaling pathway.Methods Logarithmic phase HepG2 cells were divided into control group,low,medium,and high concentration groups of isorhy(32.5,65.0,130.0 μmol·L-1),and the high concentration group of isorhy(130 μmol·L-1)+EGFR activator NSC228155(2 μmol·L-1).Logarithmic phase 5-FU-resistant hepatocellular carcinoma cell line HepG2/5-FU was divided into control group,isorhy(130 μmol·L-1)group,5-FU(60 μmol·L-1)group,isorhy(130 μmol·L-1)+5-FU(60 μmol·L-1)group,and isorhy(130 μmol·L-1)+5-FU(60 μmol·L-1)+NSC228155(2 μmol·L-1)group.The cells were treated with drugs for 24 hours,and the control group was not treated.EdU staining and CCK-8 were applied to detect HepG2 or HepG2/5-FU cell proliferation.Scratch experiment was applied to detect the migration of HepG2 or HepG2/5-FU cells.Western blotting was applied to detect cyclinD1,MIEN1,P-glycoprotein(P-gp),p-EGFR,and p-FAK proteins in cells.Results Compared with the control group,the EdU positive rate,A450 value,scratch healing rate,CyclinD1,MIEN1,p-EGFR,and p-FAK protein expression of HepG2 cells in isorhy group were reduced,in a concentration-dependent manner(P＜0.05).Compared with isorhy(130 μmol·L-1)group,the EdU positive rate,A450 value,scratch healing rate,CyclinD1,MIEN1,p-EGFR,and p-FAK protein expression of HepG2 cells in isorhy(130.0 μmol·L-1)+NSC228155 group were increased(P＜0.05).Compared with control group,the EdU positive rate,scratch healing rate,A450 value,P-gp,p-EGFR,and p-FAK protein expression of HepG2/5-FU cells in the 5-FU group were reduced(P＜0.05).Compared with the isorhy group or 5-FU group,the EdU positive rate,A450 value,scratch healing rate,P-gp,p-EGFR,and p-FAK protein expression of HepG2/5-FU cells in the isorhy+5-FU group were reduced(P＜0.05).Compared with the isorhy+5-FU group,the positive rate of EdU,A450 value,scratch healing rate,P-gp,p-EGFR,and p-FAK protein expression in HepG2/5-FU cells in the isorhy+5-FU+NSC228155 group were increased(P＜0.05).Conclusion Isorhy may inhibit proliferation,migration,and reduce 5-FU resistance of HepG2 cells by blocking the EGFR/FAK pathway.
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