过氧化物酶体增殖因子激活受体γ介导5-烯丙基-7-二氟亚甲基白杨素诱导人卵巢癌CoC1细胞凋亡的研究

目的 探讨5-烯丙基-7-二氟亚甲基白杨素(ADFMChR)诱导人卵巢癌CoC1细胞凋亡作用的分子机制.方法 采用MTT法测定ADFMChR对卵巢癌CoC1细胞系细胞增殖抑制作用;DNA琼脂糖凝胶电泳证实ADFMChR诱导CoC1细胞凋亡作用;Western印迹检测ADFMChR对CoC1细胞过氧化物酶体增殖物激活受体(PPARγ)、Bcl-2及Bax蛋白表达的影响.结果 MTT法显示ADFMChR呈剂量依赖性抑制CoC1细胞增殖作用,IC50值为7.76 μmol/L;ADFMChR(30.0 μmol/L)处理CoC1细胞48 h的DNA琼脂糖凝胶电泳呈现典型"梯形"条带,加用PPARγ阻断剂(GW9662)后条带消失;Western印迹分析ADFMChR作用CoC1细胞后PPARγ和Bax蛋白表达上调,而Bcl-2蛋白表达下调(P<0.05或P<0.01);GW9662具有阻断ADFMChR对Bcl-2的下调和Bax上调作用.结论 ADFMChR通过活化PPARγ,进而减少Bcl-2/Bax比例,诱导CoC1细胞凋亡.

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 and growth inhibition of CoC1 cells treated with ADFMChR was respectively measured using(3,4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide(MTT)colorimetric assay and clone-formation in soft agar assay. The apoptosis of CoC1 cells induced by ADFMChR was determined by DNA agarose gel electrophoresis assay. Effect of ADFMChR on PPAR-γ, 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 IC50 was 7. 76 μmol/L The ladder-shaped band could be shown in DNA agarose gel electrophoresis after treatment with ADFMChR at 30. 0 μmol/L for 48 h and the ladder-shape band disappeared with GW9662. Western blot analysis showed that expression of PPAR-γ and Bax proteins were up-regulated and protein levels of Bcl-2 were depressed after treatment with ADFMChR in a concentration-dependent fashion. However,the effects of ADFMChR on Bcl-2 and Bax proteins were blocked or diminished in the presence of GW9662. Conclusions The effect of ADFMChR on induction of apoptosis in CoC1 cells may be mediated by activation of PPAR-γ, sequentially accompanied by reducing of protein levels of Bcl-2 and increasing of Bax expression.