[Induction of apoptosis of human ovarian cancer CoC1 cells by 5-allyl-7-gen-difluoromethylenechrysin through activation of peroxisome-proliferator activated receptor-gamma].

OBJECTIVE

To investigate induction of apoptosis of human ovarian cancer CoC1 cells by 5-Allyl-7-Gen-Difluoromethylenechrysin (ADFMChR) in vitro, and its molecular mechanism.

METHODS

The proliferative inhibition of CoC1 cells treated with ADFMChR was measured using (3, 4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) colorimetric assay. The apoptosis of CoC1 cells induced by ADFMChR was determined by DNA agarose gel electrophoresis assay and flow cytometry using PI staining. Effect of ADFMChR on PPARgamma, NF-kappaB, Bcl-2, Bax protein expression level of CoC1 cells was detected by Western blotting.

RESULTS

The proliferation of CoC1 cells could be significantly inhibited by ADFMChR in a dose-dependent manner, The IC(50) was 7.76 micromol/L. ADFMChR significantly induced apoptosis in a concentration-dependent, the rate of apoptosis was 33.07% and 73.70% respectively after treatment with 10.0, 30.0 micromol/L of ADFMChR for 48 h, which was higher than either the control group (21.70%, 40.00%) at the same concentration ChR-treated cells. The ladder-shaped band could be shown in DNA agarose gel electrophoresis after treatment with ADFMChR at 30.0 micromol/L for 48 h and the ladder-shape band disappeared with GW9662. Western Blot analysis shown that expression of PPARgamma and Bax proteins were upregulation and protein levels of NF-kappaB and Bcl-2 were depress after treatment with ADFMChR in a concentration-dependent.

CONCLUSION

The effect of ADFMChR on induction of apoptosis in CoC1 cells may be mediated by activation of PPARgamma, sequentially accompanied by reducing of protein levels of NF-kappaB and Bcl-2 and increasing of Bax expression.