新藤黄酸联合阿霉素诱导人乳腺癌MCF-7细胞凋亡的研究

Apoptosis-inducing Effect of Gambogenic Acid Combined with Adriamycin on Human Breast Cancer MCF-7 Cells

目的研究新藤黄酸（gambogenic acid,GNA）与阿霉素（adriamycin,ADM）联合作用对人乳腺癌细胞MCF-7的影响。方法体外培养人乳腺癌细胞MCF-7细胞株,采用甲基噻唑基四唑（methyl thiazolyl tetrazolium,MTT）检测细胞存活率,倒置显微镜观察细胞形态变化;4′,6-二脒基-2-苯基吲哚（4′,6-diamidino-2-phenylindole,DAPI）细胞核染色实验观察GNA与ADM单独与联合作用对乳腺癌细胞MCF-7凋亡的影响;膜联蛋白Ⅴ（annexinⅤ,AV）-异硫氰酸荧光素（fluorescein isothiocyanate,FITC）/碘化丙啶（propidium iodide,PI）双染流式细胞仪检测给药后乳腺癌细胞MCF-7凋亡率。结果 MTT检测结果显示,在GNA浓度为0.125～4μmol/L和ADM浓度在0.5～16μmol/L范围内,人乳腺癌细胞MCF-7的细胞存活率随GNA和ADM剂量增加而降低,且联合用药组细胞存活率低于单独用药组。倒置显微镜和荧光显微镜下,与对照组比较,细胞明显缩圆变小,染色细胞核破碎,呈现凋亡状态,而联合用药组凋亡细胞数量明显高于单独用药组。AV-FITC/PI双染经流式细胞仪检测发现,GNA与ADM单独和联合作用均诱导MCF-7细胞凋亡,与单独用药组比较,联合用药组细胞凋亡率显著增加（P〈0.01）。结论 GNA与ADM单独和联合作用均能诱导人乳腺癌细胞MCF-7细胞凋亡,且两者联合作用可产生协同效应,其作用机制可能与诱导细胞凋亡相关。

Objective To study the apoptosis-inducing effect of gambogenic acid（GNA）combined with adriamycin（ADM）on human breast cancer MCF-7cells.Methods Human breast cancer MCF-7cells were cultured invitro.Cell viability was determined by MTT assay;cell morphology was observed by inverted microscopy;nuclear staining with 4′,6-diamidino-2-phenylindole was used to assess the effect of GNA and ADM used alone or together on the apoptosis of MCF-7cells;annexin V（AV）-fluorescein isothiocyanate（FITC）/propidium iodide（PI）staining and flow cytometry were used to measure the apoptosis rate of MCF-7cells treated with GNA and/or ADM.Results The MTT assay showed that the viability of MCF-7cells decreased as the concentrations of GNA and ADM increased within 0.125-4μmol/L and0.5-16μmol/L,respectively,and the cell viability was lower when GNA and ADM were used together than when they were used alone.The inverted microscopy and fluorescence microscopy showed that,compared with control cells,treated cells became markedly round and small,had broken nuclei,and were apoptotic,and the number of apoptotic cells was larger when both GNA and ADM were used than when they were used alone.AV-FITC/PI staining and low cytometry showed that GNA and/or ADM induced the apoptosis of MCF-7cells,and the apoptosis rate increased significantly when GNA and ADM were used together than when they were used alone（P〈0.01）.Conclusion The single and combined use of GNA and ADM can induce the apoptosis of human breast cancer MCF-7cells,and the combination of two drugs has a synergistic effect.The mechanism may be related to inducing cell apoptosis.