Apoptosis in HepG2 cells induced by Pteris semipinnata extract 5F involves p53 activation and VEGF inhibition.

Objective To observe the effect of Pteris semipinnata extract,ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid(5F),on human hepatoma HepG2 cells about the proliferation and to investigate the role of p53,VEGF,and Caspase-3 in 5F-induced HepG2 cell apoptosis.Methods MTT Assay was used to determine the effect of 5F on proliferation of HepG2 cells,and cytoplasmic mono-and oligonucleosomes of HepG2 cells treated with 5F was assessed with Cell Death Detection ELISA Kit for determining whether 5F can induce apoptosis.Apoptotic nuclear morphology was assessed using Hoechst/PI assay.The effect on the expression levels of p53 and VEGF protein in HepG2 cells was detected by Western blotting analysis,and Caspase-3 activity was measured by a Caspase-3 Colorimetric Assay Kit.Results The cytotoxicity of 5F on HepG2 cells was elevated with 5F concentration increasing.Cytoplasmic mono-and oligonucleosomes were induced by 5F in HepG2 cells,and the induction was in a concentration-dependent manner.After treatment with 5F,the apoptotic change such as chromatin condensation was confirmed by Hoechst/PI staining.The level of p53 in nucleus was significantly elevated in HepG2 cells after 5F treatment,but the level of VEGF in cytosol was decreased.Moreover,the activity of Caspase-3 increased in a concentration-dependent manner.Conclusion 5F mediated apoptosis involves p53 and Caspase-3 activation,VEGF down regulation in HepG2 cells.5F might have a therapeutic value against human cancer cell lines and especially on hepatocellular carcinoma(HCC) cells.