新藤黄酸诱导卵巢癌A2780细胞凋亡的作用研究
Effect of Gambogenic Acid on the Apoptosis of Ovarian Cancer Cell Line A2780
  
DOI:
中文关键词:  新藤黄酸  卵巢癌细胞A2780  线粒体凋亡
英文关键词:Gambogenic acid  Ovarian cancer cell line A2780  Mitochondrial apoptosis
基金项目:国家自然科学基金项目(81673650,81573615);安徽省自然科学研究重点项目(KJ2017A288)
作者单位
程 卉,李庆林,侯 梅,苏婧婧 安徽中医药大学科研实验中心 新安医学教育部重点实验室安徽 合肥 230038 
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中文摘要:
      目的 研究新藤黄酸对卵巢癌细胞增殖的抑制作用及其可能的机制。方法 采用MTT检测新藤黄酸对卵巢癌A2780细胞存活率的影响,采用fluo-3AM染色荧光显微镜观察新藤黄酸对A2780细胞钙离子水平的影响,采用Hoechst 33342染色荧光显微镜观察细胞的凋亡率,采用PI染色流式细胞仪检测新藤黄酸对细胞周期的影响,采用Annexin Ⅴ-FITC/PI染色流式细胞仪检测新藤黄酸对细胞凋亡率的影响,采用Western blot检测凋亡相关蛋白表达的影响。结果 MTT检测结果表明,新藤黄酸对卵巢癌细胞A2780的增殖具有抑制作用,且抑制作用具有明显的剂量依赖性;fluo-3AM染色荧光显微镜下观察,可见各浓度新藤黄酸均能升高A2780细胞内钙离子水平,且呈现浓度依赖关系;PI染色流式细胞仪检测结果表明,新藤黄酸能阻滞A2780细胞于G0/G1期;Hoechst 33342染色和Annexin Ⅴ-FITC/PI染色流式细胞仪检测结果表明,随着新藤黄酸浓度的增加,A2780细胞凋亡率呈现增高的趋势;Western blot检测结果表明,新藤黄酸可以促进凋亡相关蛋白p53、细胞色素C以及Caspase-9表达水平升高。结论 新藤黄酸具有抑制卵巢癌A2780细胞增殖和诱导肿瘤细胞发生线粒体凋亡的作用。
英文摘要:
      Objective To investigate the inhibitory effect of gambogenic acid on the proliferation of ovarian cancer cells and its possible mechanism. Methods MTT assay was used to measure the effect of gambogenic acid on the viability of ovarian cancer A2780 cells. Fluo-3/AM staining was used to observe the effect of gambogenic acid on the level of calcium ions in A2780 cells under a fluorescence microscope. Hoechst 33342 staining was used to observe the apoptosis rate of A2780 cells under a fluorescence microscope. PI flow cytometry was used to observe the effect of gambogenic acid on cell cycle. Annexin V-FITC/PI flow cytometry was used to observe the effect of gambogenic acid on apoptosis rate. Western blot was used to measure the expression of apoptosis-related proteins. Results MTT assay showed that gambogenic acid significantly inhibited the proliferation of ovarian cancer A2780 cells in a dose-dependent manner. Fluo-3/AM staining under the fluorescence microscope showed that gambogenic acid promoted the level of calcium ions in A2780 cells in a concentration-dependent manner. PI flow cytometry showed that A2780 cells were arrested in G0/G1 phase by gambogenic acid. Hoechst 33342 staining and Annexin V-FITC/PI flow cytometry showed that the apoptosis rate of A2780 cells gradually increased with the increase in the concentration of gambogenic acid. Western blot showed that gambogenic acid promoted the expression of the apoptosis-related proteins P53, cytochrome C, and Caspase-9. Conclusion Gambogenic acid can inhibit the proliferation of ovarian cancer A2780 cells and induce mitochondrial apoptosis.
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