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

Induction of Apoptosis of human ovarian cancer CoC1 cells by 5-Allyl-7-Gen-Difluoromethylenechrysin through activation of peroxisome-proliferator activated receptor-gamma

作　　者：李华珍[1] 曹建国[2] 邓宇傲[1] 许金华[3] 谢宛玉[1] 

机构地区：[1]南华大学附属第一医院妇产科,衡阳421001 [2]南华大学肿瘤研究所 [3]南华大学生命科学院生化中心

出　　处：《中华医学杂志》2007年第41期2914-2918,共5页National Medical Journal of China

基　　金：湖南省科技厅科研项目(06FJ3125)

摘　　要：目的探讨5-烯丙基-7-二氟亚甲基白杨素(ADFMChR)通过活化过氧化物酶体增殖因子激活受体(PPARγ)诱导人卵巢癌细胞凋亡作用的分子机制。方法采用 MTF 法测定 ADFMChR对卵巢癌(CoCl)细胞系细胞增殖抑制作用;碘化丙啶染色流式细胞术(FCM)检测 ADFMChR 诱导CoCl 细胞凋亡程度;DNA 琼脂糖凝胶电泳证实 ADFMChR 诱导 CoCl 细胞凋亡作用。Western 印迹检测 ADFMChR 对 CoCl 细胞 PPARγ,NF-kB,Bcl-2,Bax 蛋白表达的影响。结果 MTT 法显示ADFMChR 呈剂量依赖性抑制 CoCl 细胞增殖作用,IC_(50)值为7.76 μmol/L;FCM 分析 ADFMChR 呈剂量依赖性诱导 CoCl 细胞凋亡,ADFMChR(10.0,30.0 μmol/L)处理 CoCl 细胞48 h,凋亡率分别为33.07%和73.70%,均高于相应浓度白杨素诱导凋亡率(21.70%、40.00%);ADFMChR(30 μmol/L)处理 CoCl 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 以剂量依赖方式上调 CoCl 细胞 PPARγ和 Bax 蛋白表达,下调NF-kB 和 Bcl-2蛋白表达。结论 ADFMChR 通过活化 PPARγ抑制 NF-kB 和 Bcl-2表达,上调 Bax,诱导 CoCl 细胞凋亡。Objective To investigate induction of apoptosis of human ovarian cancer CoC1 cells by 5-Allyl-7-C, en-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 PPARγ, NF-κB, 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 dosedependent manner, The IC50 was 7.76 μmol/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 μmo[/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 μLmo[/L for 48 h and the ladder-shape band disappeared with GW9662. Western Blot analysis shown that expression of PPARγ and Bax proteins were upregulation and protein levels of NF-KB and Bcl-2 were depress after treatment with ADFMChR in a concentrationdependent. Conclusion The effect of ADFMChR on induction of apoptosis in CoC1 cells may be mediated by activation of PPARγ , sequentially accompanied by reducing of protein levels of NF-κB and Bcl-2 and increasing of Bax expression.

关 键 词：卵巢肿瘤 白杨素 PPARΓ 凋亡 

分 类 号：R737.31[医药卫生—肿瘤]