Crocin promotes apoptosis and autophagy of human hepatoma HepG2 cells through PI3K/Akt/mTOR pathway
Objective To study the effect and mechanism of Crocin on apoptosis and autophagy of human hepatoma HepG2 cells.Methods Different concentrations of Crocin(0,0.1,0.2,0.4,0.8,1.6,3.2 mmol/L)were used to intervene HepG2 cells in logarithmic growth phase for 48 h,the proliferation inhibition rate was detected by MTT method,and the half-inhibitory concentration(IC50)was calculated.The IC50 was used as the drug concentration in subsequent experiments.The blank control group,Crocin group,Crocin+IGF-1(PI3K activator)group and Crocin+LY294002(PI3K inhibitor)group were set up.Forty-eight hours after drug intervention,the proliferation inhibition rate and the apoptosis rate were respectively detected by MTT or Annexin V-FITC/PI double staining method.The autophagosomes was observated after transfection of GFP-LC3 fluorescent plasmid into HepG2 cells.The expression of Cleaved Caspase-3,Cleaved Caspase-9,LC3,Beclin-1,PI3K,p-PI3K,Akt,p-Akt,mTOR,p-mTOR were detected by Western blot.Results The inhibitory effect of Crocin on HepG2 cell proliferation was dose-dependent,which IC50 value was 0.78 mmol/L.Compared with the blank control group,the proliferation inhibition rate,apoptosis rate and the autophagosomes number of HepG2 cell in Crocin group were significantly increased(P＜0.05);the expressions of Cleaved Caspase-3,Cleaved Caspase-9,LC3-I,LC3-Ⅱ,Beclin-1 and the ratio of LC3-Ⅱ/LC3-I were significantly increased(P＜0.05);the phosphorylation level of PI3K,Akt,mTOR(p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR)were significantly decreased(P＜0.05).IGF-1 could significantly reverse the above-mentioned effects of Crocin on HepG2 cells,while LY294002 could significantly enhance the above-mentioned effects of Crocin on HepG2 cells,the differences were significant(P＜0.05).Conclusion Crocin can promote the apoptosis and autophagy of human hepatoma HepG2 cells,which mechanism may be related to the inhibition of PI3K/Akt/mTOR pathway.
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