自组装纳米短肽基质在肿瘤细胞三维培养中的应用

Application of self-assembling nano-oligopeptide in three-dimensional culture of cancer cell

目的研究人肺癌细胞A549在自组装纳米短肽RADA16三维基质材料中培养时的细胞学行为,揭示肿瘤细胞在三维环境与传统二维环境中培养的差异。方法用透射电子显微镜对材料进行表征;MTT检测细胞对5-氟尿嘧啶的敏感性;黏附性实验分析二维及三维培养的细胞对胞外基质蛋白(Ⅳ型胶原、层粘连蛋白、纤粘连蛋白)黏附率的差异。结果RADA16形成了类似体内细胞外基质的纳米纤维网络。A549在三维基质中的状态较好,随着时间增加、抗药性增加,药物敏感性降低。对不同的胞外基质蛋白黏附率揭示,与二维培养的细胞相比,RADA16三维培养的细胞克隆对不同胞外基质蛋白的黏附性有显著性差异。结论细胞在RADA16三维培养中,细胞的生长、形貌、药敏性及对胞外基质黏附性等细胞行为与二维培养的细胞具有显著性差异,展示了纳米短肽材料在细胞三维培养研究中的应用前景。

Objective To investigate the cellular behaviors of human lung cancer cells （A549） cultured in three-dimensional （3D） self-assembling nanofiber peptide RADA16 matrix and to reveal the differences of tumor cell behaviors in 3D and two-dimensional （2D） cultures. Methods The structure of the peptide was tested with transmission electron microscopy. The sensitivity of the cells to 5-Fu was detected by MTT assay. The adhesion experiment was employed to analyze the differences of adhesion ability for extra-cellular matrix protein （lVcollagen, laminin, fibronectin） of cells cultured under 2D and 3D conditions. Results RADA16 formed nanofiber networks which were similar to the extra-cellular matrix. A549 grew well in the 3D peptide matrix, and with the culture time increased, its resistance to drug increased whereas its sensitivity to drug reduced. Regarding the adhesion ability to different extra-cellular matrix, there were significant differences between cells cultured in 3D RADA16 matrix and cells in 2D culture. Conclusion Under 3D RADA16 culture, cellular behaviors such as the growth, morphology, drug sensitivity and adhesion to extra-cellular matrix are significantly different from those in 2D culture. This shows the prospects of self-assembling nano-oligopeptide used in cell 3D culture.