人乳腺癌细胞MDA-MB-231三维培养模型的构建

Construction of Three-dimensional Culture Model of Human Breast Cancer Cells MDA-MB-231

本研究旨在摸索一种体外构建人乳腺癌细胞MDA-MB-231三维(3D)培养的新方法,以期建立一种能够模拟体内肿瘤细胞真实形态的理想模型。利用该培养模型比较分析MDA-MB-231与正常乳腺上皮细胞MCF-10A 3D形态发生过程中极性形成的差异,主要是采用3D腺泡免疫荧光染色技术对顶部极性标记GM130和底部极性标记Laminin-5的定位进行比较研究,结果发现在3D培养10 d,MDA-MB-231形成了一种近似体内的球形结构,confocol层切MDA-MB-231球体的赤道轴发现,相对于MCF-10A形态发生形成的极性空腔腺泡,MDA-MB-231形成的是一种极性紊乱的实心球结构,其顶部极性标记GM130呈无规则分布、底部极性标记Laminin-5向球体中心内渗,这些结果与体内乳腺癌细胞去极性的特征相吻合,表明该3D培养模型可为深入阐明乳腺癌细胞MDA-MB-231恶化转移及抗癌药物作用机制等研究提供可靠的实验模型。

The purpose of this study is to explore a new method for the construction of MDA-MB-231 three dimension cultures in vitro, to establish an ideal model which can simulate the real morphology of tumor cells in vivo. The differences in polarity formation between MDA-MB-231 and normal breast epithelial cells MCF-10 a during morphology were compared and analyzed using this new 3 D culture model on Day 10, which were monitored by immunostaining analysis for GM130 and laminin-5. We found that MCF-10 A cells developed polarized structures, with GM130 exhibiting an apical location toward the lumen and a clearly defined basement membrane containing laminin-5, while MDA-MB-231 cells developed into solid spheres, with disordered GM130 orientation and excessive deposition of laminin-5. These results are consistent with the characteristics of the depolarization of breast cancer cells in vivo, and indicate that the new 3 D culture model can provide a reliable experimental model for the in-depth study of the progression of MDA-MB-231 and the mechanism of anticancer drugs in breast cancer cells.