乳腺癌细胞三维培养模型构建及其药物敏感性研究  

Fabrication of Three Dimensional Cell Culture Model of Breast Adenocarcinoma and Detection of Its Drug Resistance

作　　者：姜昊声[1] 曹燕[1] 吴绍秋[1] 刘冰妍[1] 贾一平[1] 王振磊[1] 尚鸣异[1] 

机构地区：[1]上海交通大学医学院附属同仁医院,200072

出　　处：《实用癌症杂志》2016年第6期871-874,共4页The Practical Journal of Cancer

摘　　要：目的观察乳腺癌MCF-7细胞在3D与2D培养模式下对阿霉素的敏感性。方法应用模板印制凝胶微孔支架技术建立体外3D细胞培养模型,通过倒置显微镜等方法观察3D细胞球的生长特性;采用Alamar Blue法获得不同培养模式下药物阿霉素作用的细胞抑制率;采用流式细胞术比较2D培养和3D培养细胞周期、细胞凋亡的差异。结果3D与2D培养相比,G-1/G-0期胞增多,S期、G2/M期细胞减少,细胞周期受阻滞,具有统计学差异（P〈0.05）;活细胞减少,早期凋亡、晚期凋亡、坏死细胞比例增大（P〈0.05）。阿霉素作用3 d后,3D培养细胞的IC50值为23.77μg/ml[95%CI 15.80-35.77],2D培养细胞的IC-（50）值为0.46μg/ml（95%CI 0.28-0.75）,MCRI为51.7。结论 3D培养MCF-7细胞在细胞周期、细胞凋亡等方面与2D培养细胞存在明显差异,可能是导致乳腺癌多细胞耐药的重要原因。Objective To observe the changes of sensitivity to doxorubicin on breast adenocarcinoma MCF-7 cells cultured as 3 dimensional model or monolayer. Methods MCF-7 cells were cultured as 3 dimensional model using agarose scaffolds with highly ordered micro-wells. The spheroid growth characterization was observed by invert microscope. The sensitivity to doxorubicin on MCF-7 spheroids or monolayers was determined by Alamar Blue assay respectively. Cell cycle and apoptotic rate were detected by flow cytometry. Results For 3D spheroid culture,the proportion of cells in G0- G1 phase was higher,while the proportion of cells in S phase and G2-M phase were lower than the 2D culture（ P〈0. 05）. The cell viability declined in 3D culture than the 2D control,and the proportion of cells in the early-apoptotic,late-apoptotic phase increased significantly（ P〈0. 05）. After 3days of DOX treatment,the IC（50） value obtained for spheroid culture was 23. 77 μg / ml [95% CI 15. 80 - 35. 77],relative to 0. 46μg /ml（ 95% CI 0. 28 - 0. 75） for the monolayer culture. The MCRI value was 51. 7. Conclusion There are significant differences in cell cycle and apoptotic rate between MCF-7 spheroids and monolayer,which may be the important causes of the multicellular resistance of breast carcinoma.

关 键 词：乳腺癌 肿瘤多细胞球 3D培养 阿霉素 抗药性 

分 类 号：R737.9[医药卫生—肿瘤]