芹菜素增敏TRAIL诱导卵巢癌CoC1细胞凋亡

Sensitization of Human Ovarian Cancer CoC1 Cells to TNF-Related Apoptosis-Inducing Ligand(TRAIL)-Induced Apoptosis by Apigenin

目的研究芹菜素(API)抑制NF-κB活性,增强肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导人卵巢癌CoC1细胞凋亡的作用。方法体外培养CoC1细胞。碘化丙啶(PI)染色流式细胞术(FCM)定量分析细胞凋亡率(亚二倍体DNA含量细胞百分率)。DNA琼脂糖凝胶电泳观察细胞DNA梯形条带。Western blot检测细胞蛋白的表达。结果 API(20μmol/L)和TRAIL(20 ng/mL)以及两者合用48 h,CoC1细胞凋亡率分别是8.83%±2.33%、8.32%±2.80%和69.5%±4.65%;API(20μmol/L)预孵育4 h后,TRAIL(20 ng/mL)处理CoC1细胞44h,展示出典型DNA梯形条带图谱。API(20μmol/L)以时间依赖的方式降低CoC1细胞IκBα蛋白磷酸化水平和抑制NF-κB(p65)蛋白表达。结论亚细胞毒性浓度的API具有增强TRAIL诱导人卵巢癌CoC1细胞凋亡作用,其作用机制与抑制NF-κB活性有关。

Objective To investigate whether apigenin（API）enhance the apoptosis induced by TNF-related apoptosis-inducing ligand（TRAIL）via inhibiting activities of NF-κB in human ovarian cancer CoC1 cell line.Methods Human ovarian cancer CoC1 cells were cultured in vitro.Apoptotic rate was determined by flow cytometry using PI fluorescence staining.The characteristic features of cell apoptosis was examined using DNA agarose gel electrophoresis.The expressions of protein were analyzed by Western blot.Results Flow cytometry analysis with PI stainning indicated that apoptotic rates in human ovarian cancer CoC1 cells by API（20 μmol/L）or TRAIL（20 ng/mL）or both for 48 h were 8.83%±2.33%,8.32%±2.80% and 69.50%±4.65%,respectively.The ladder band could be shown in DNA agarose gel electrophoresis by pretreated with API（20 μmol/L）for 4 h then with TRAIL（20 ng/mL）for 44 h.Western blot analysis indicated that API inhibited expression of NF-κB（p65）protein and reduced the phosphorylated level of IκBα protein in CoC1 cells,in a time-dependent manner.Conclusion API at subtoxic concentration potentiates the induction of apoptosis by TRAIL,and is associated with inhibiting activities of NF-κB in human ovarian cancer CoC1 cell line.