Drug resistance of sorafenib seriously affects the treatment effect of late-stage hepatocellular carcinoma（HCC） patients.However,the precise mechanism of resistance to sorafenib remains unclear.Therefore,to obtain a deep understand of sorafenib resistance mechanisms and find potential therapeutic targets are very important for improving the clinical prognosis of HCC patients.In this study,a label-free quantitative proteomics method was performed to investigate the proteins differentially expressed between HepG2 and the sorafenib-acquired resistance HepG2（HepG2-R） cells.In total,84 differential expressed proteins were identified between the two cell lines.Bioinformatics analysis results demonstrated the dysregulated metabolic processes have a significant impact on the drug resistance of HepG2-R cells.Among them,the expression of Microsomal glutathione S-transferase 1（MGST1） in two cell lines was further confirmed by western blot method.Moreover,colony formation assay and trypan blue dye assay results revealed that MGST1 is closely connected with the sorafenib resistance of HepG2-R cells,and the knockdown of MGST1 increased the sensitivity of sorafenib resistance HepG2-R cells to sorafenib treatment.In conclusion,these results lay a foundation for deciphering the mechanism for HCC sorafenib resistance and present a possibility of MGST1 serving as a therapeutic target for the treatment of sorafenib resistance HCC.