OBJECTIVE To investigate the regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib.METHODS Using human liver cancer cell Huh7 as subject,the lenvatinib-resist cell model(Huh7-LR)was generated by the low-dose gradient method combined with long-term administration.The sensitivity of parental cell Huh7 and drug-resistant cell Huh7-LR to lenvatinib was detected by using CCK-8 assay and flow cytometry.Western blot assay and GFP-mCherry-LC3 plasmid transfection were performed to detect the expression levels of autophagic protein Beclin-1,autophagic adapter protein sequestosome 1(p62),microtubule-associated protein 1 light chain 3(LC3)and autophagic level.Furthermore,an autophagy activation model was constructed by cell starvation,the protein expression of p62 and autophagy level were detected by using Western blot assay and GFP-mCherry-LC3 plasmid transfection,and the effect of autophagy activation on the sensitivity of Huh7-LR cells to lenvatinib was detected by flow cytometry.RESULTS Compared with parental cells,the drug resistance index of Huh7-LR cells was 6.2;protein expression of p62 was increased significantly,while apoptotic rate,protein expression of Beclin-1 and LC3Ⅱ/LC3Ⅰ ratio were all reduced significantly(P＜0.05 or P＜0.01);the level of autophagy was decreased to some extent.Autophagy activation could significantly increase the protein expression of p62 in Huh7-LR cells(P＜0.05)and autophagy level,and significantly increase its apoptotic rate(P＜0.05).CONCLUSIONS Autophagy is involved in lenvatinib resistance,and activating autophagy can reverse the resistance of liver cancer cells to lenvatinib to some extent.
autophagylenvatinibliver cancer celldrug resistance
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