ABSTRACT
Objective::To investigate the effect of bergapten on the apoptosis of HepG2 and Hep3B cells through phosphatidylinositol 3 kinase (PI3K)/protein kinase B(Akt) pathway. Method::Bergamot (5, 50, 200 μmol·L-1) groups and blank group were set up. The effect of bergapten at different concentrations on proliferation of HepG2 and Hep3B cells for 24, 48 h were detected by thiazolyl blue(MTT) assay. Apoptosis was detected by Annexin V-FITC/propidium iodide double staining. Quantitative real-time fluorescence reverse transcriptional polymerase chain reaction (Real-time PCR) and Western blot assay were used to detect relevant mRNA and proteins expressions. The clone formation rate and the effect of HepG2 and Hep3B cells in each group were evaluated by plate cell clone formation. Result::MTT assay showed that bergapten could significantly inhibit the proliferation activity of HepG2 and Hep3B cells in a time-dependent manner. Flow cytometry analysis showed that bergapten in 200 μmol·L-1 concentration groups had significant pro-apoptotic effect on HepG2 and Hep3B cells after 48 h (P<0.05). Western blot results showed that bergamolactone could up-regulate the protein expressions of Caspase-3, Caspase-8 (P<0.05), and down-regulate protein expressions of B-lymphocytoma-2 (Bcl-2), PI3K (P<0.05). Real-time PCR results showed that mRNA expressions of PI3K and Akt were decreased(P<0.05). The results of plate cell clone formation experiment showed that with the increase of the concentration of bergamolide, the cell clone formation rate of each group showed a decreasing trend, particularly in 200 μmol·L-1 concentration group (P<0.05). Conclusion::Bergapten can inhibit the proliferation of HepG2 and Hep3B cells, which may be induced through the PI3K/Akt signaling pathway.