摘要
目的研究新藤黄酸(gambogenic acid,GNA)诱导人结肠癌HCT116细胞凋亡的机制。方法采用不同浓度的GNA作用细胞24h,对照组是相同浓度GNA加内质网应激抑制剂4-苯基丁酸(4-phenylbutyric acid,4-PBA)共同作用HCT116细胞24h。采用甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)染色测定两组细胞增殖抑制率的差异,采用吖啶橙(acridine orange,AO)和溴化乙锭(ethidium bromide,EB)染色观察细胞的形态学变化,采用膜联蛋白Ⅴ(AnnexinⅤ,AV)-异硫氰酸荧光素(fluorescein isothiocyanate,FITC)/碘化丙碇(propidium iodide,PI)双重染色检测细胞凋亡率。结果内质网应激抑制剂4-PBA可以缓解GNA对HCT116细胞增殖的抑制作用;AO/EB染色后荧光显微镜观察发现GNA作用的细胞具有凋亡特征;流式细胞仪检测显示4-PBA可降低HCT116细胞的凋亡率。结论 GNA能抑制人结肠癌细胞HCT116增殖,诱导细胞凋亡,其诱导细胞凋亡的作用可能与内质网应激途径有关。
Objective To investigate the mechanism by which gambogenic acid （GNA） induces the apoptosis of human colon cancer HCTll6 cells. Methods Experimental HCTll6 cells were treated with 2.5, 5.0, and 7.5 μmol/L GNA for 24 h, while control HCTll6 cells were treated with 2.5, 5.0, and 7.5 μmol/L GNA plus 5 μmol/L endoplasmic reticulum stress （ERS） inhibitor 4-phenylbutyric： acid （4-PBA） for 24 h. The cellular proliferation inhibition rate was measured by methyl thiazolyl tetrazolium assay; the morphological changes of HCTll6 cells were observed by acridine orange （AO）/ethidium bromide （EB） staining under a fluorescence microscope; the cell apoptosis rate was determined by flow cytometry with annexin V-fluorescein isothiocyanate （FITC）/propidium iodide （PI） double staining. Results The ERS inhibitor 4-PBA reduced the inhibitory effect of GNA on the proliferation of HCTll6 cells. The AO/EB staining indicated the characteristics of apoptosis in GNA-treated HCT116 cells under the fluorescence microscope. The flow cytometry with annexin V-FITC/PI double staining showed that 4-PBA reduced the apoptosis rate of GNA-treated HCT116 cells. Conclnsion GNA can inhibit proliferation and induce apoptosis in HCT116 cells, and it might induce cell apoptosis through ERS.
出处
《安徽中医药大学学报》
CAS
2014年第1期63-66,共4页
Journal of Anhui University of Chinese Medicine
基金
安徽省自然科学基金项目(11040606M190)
国家自然基金项目(81173600)
