乳腺癌细胞三维培养模型构建及其药物敏感性研究

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AlamarBlue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，G1／G0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IC50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。

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

Objective To observe the changes of sensitivity to doxorubicin on breast adenocarcinoma MCF -7 cells cul-tured 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 doxoru-bicin on MCF-7 spheroids or monolayers was determined by AlamarBlue assay respectively .Cell cycle and apoptotic rate were de-tected by flow cytometry.Results For 3D spheroid culture,the proportion of cells in G0 ~G1 phase was higher,while the propor-tion 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 3 days of DOX treatment,the IC50 value obtained for spheroid culture was 23.77μg/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 differ-ences in cell cycle and apoptotic rate between MCF-7 spheroids and monolayer ,which may be the important causes of the multi-cellular resistance of breast carcinoma .