Ethyl acetate extract of Hedyotis diffusa Willd induce AML cells via PI3K/Akt/p53 signaling pathway
Objective It is to evaluate the regulating effect of ethyl acetate extract of Hedyotis diffusa Willd(EAE-HDW)on PI3K/Akt/p53 signaling pathway,and to explore its mechanism of inhibiting the proliferation and inducing ap-optosis of acute myeloid leukemia type M2(AML-M2)cell line Kasumi-1.Methods The EAEHDW was prepared by ex-tracting from Hedyotis diffusa Willd with ethyl acetate,and the sample purity was detected by high performance liquid chro-matography tandem mass spectrometry device.Blank groups were set up,the Kasumi-1 cell line at logarithmic growth phase were treated with 0.02 mg/mL,0.04 mg/mL,0.06 mg/mL,0.08 mg/mL,0.10 mg/mL EAEHDW for different time,and their survival rates were detected by MTT assay,their apoptosis was detected by Annexin V/PI flow cytometry after 24 h of treatment with different concentrations of EAEHDW,and the expressions of Caspase family proteins and PI3K/Akt/p53 sig-naling pathway proteins in the cells were detected by Western blot after 24 of treatment with different concentrations of EAE-HDW(0.02 mg/mL,0.04 mg/mL,0.06 mg/mL).Results Compared with the blank group,the survival rate of Kasumi-1 was significantly decreased and the apoptotic rate was significantly increased after treatment with different doses of EAE-HDW,the cell survival rate decreased more significantly with the prolongation of the treatment time and the increase of the drug concentration,and the apoptosis rate increased more significantly with the increase of the drug concentration,and the differences were all statistically significant(all P<0.05).The relative expressions of poly ADP ribose polymerase(PARP),Caspase-3,Caspase-8,Caspase-9,cytochrome C(Cyto-C),and p53 proteins were significantly increased in Kasumi-1 cells after treatment with different concentrations of EAEHDW(all P<0.05),and the relative expressions of Akt,phosphorylated Akt(p-Akt),murine double microchromosome 2(MDM2),phosphorylated MDM2(p-MDM2)pro-teins were significantly decreased(all P<0.05).Conclusion EAEHDW can significantly inhibit the proliferation of mye-loid leukemia Kasumi-1 cells and induce their apoptosis,the mechanism of which may be related to influencing the expres-sion of Caspase family proteins and inhibiting PI3K/Akt/p53 signaling pathway to lead to p53 activation.
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