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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 the molecular mechanism of 5-allyl-7-difluoromethylenechrysin (ADFMChR) inducing apoptosis of human ovarian cancer cells by activating peroxisome proliferator-activated receptor (PPARγ). Methods The inhibitory effect of ADFMChR on the cell proliferation of ovarian cancer (CoCl) cells was determined by MTT assay. The apoptosis of CoCl cells induced by ADFMChR was detected by propidium iodide staining (FCM). The DNA fragmentation was detected by DNA agarose gel electrophoresis. Apoptosis. The effect of ADFMChR on the protein expression of PPARγ, NF-κB, Bcl-2 and Bax in CoCl cells was detected by Western blotting. Results MTT assay showed that ADFMChR could inhibit the proliferation of CoCl cells in a dose-dependent manner with the IC 50 value of 7.76 μmol / L. FCM showed that ADFMChR induced apoptosis of CoCl cells in a dose-dependent manner, and ADFMChR (10.0 and 30.0 μmol / L) The apoptotic rate was 33.07% and 73.70% at 48 h, respectively, higher than those induced by chrysin at 21.70% and 40.00%, respectively. DNA fragmentation of CoCl cells treated with ADFMChR at 30 μmol / The bands disappeared after addition of PPARγ blocker (GW9662). Western blot analysis showed that ADFMChR upregulated the expression of PPARγ and Bax protein and down-regulated the expression of NF-κB and Bcl-2 protein in CoCl cells in a dose-dependent manner . Conclusion ADFMChR inhibits the expression of NF-κB and Bcl-2 by activating PPARγ, up-regulates Bax and induces apoptosis of CoCl cells.