基质硬度对2D和3D培养乳腺癌细胞摄取纳米制剂行为的-药学学报.PDF

质硬度对2D 和3D 培养乳腺癌细胞摄取纳米制剂行为的影响 王瑜，秦硕，龚涛，张志荣，符垚* ( 四川大学华西药学院，四川 成都 610041 ) 摘要：以聚苯乙烯纳米球为研究对象，研究基质硬度对二维（2D ）和三维（3D ）培养的 乳腺癌细胞摄取行为的影响。通过光催化聚合反应法制备硬度可调的聚丙烯酰胺凝胶，并在凝 胶表面共价修饰胶原以支持细胞黏附，构建2D 细胞培养模型；选择I 型鼠尾胶原构建3D 细 胞培养模型；分别接种人乳腺癌细胞MCF-7 于聚丙烯酰胺凝胶表面或混悬于胶原凝胶中进行 3D 细胞培养；20 nm 和50 nm 聚苯乙烯纳米球悬液与2D 或3D 培养的细胞共孵育，采用激光 共聚焦显微镜对细胞形态和纳米球摄取效率进行定性和定量考察。本研究发现在2D 和3D 培 养条件下，基质硬度均参与调控肿瘤细胞肌动蛋白骨架的重构并间接影响不同粒径大小的纳米 制剂的摄取行为。 关键词：基质硬度；二维/三维细胞培养；细胞形态；细胞摄取；聚苯乙烯纳米球 中图分类号: R945 文献标识码: A 文章编号: 0513-4870 （2017 ） Matrix stiffness regulates cell uptake of nanoparticles in 2D and 3D cultures of breast cancer cells WANG Yu, QIN Shuo, GONG Tao, ZHANG Zhi-rong, FU Yao* ( West China School of Pharmacy, Sichuan University, Chengdu 610041, China) Abstract: Fluorescent polystyrene nanospheres (PS) were selected as model nanoparticles to explore the impact of substrate stiffness on cell uptake of nanoparticles in breast cancer cells. Polyacrylamide (PAA) gels with varying stiffness were prepared by photopolymerization, and type I rat tail collagen was covalently conjugated on the surface of PAA gels to facilitate cell adhesion. Type I rat tail collagen was also used to fabricate collagen gels for 3D cell culture. Human breast cancer cell line MCF-7 cells were subjected to 2D culture on PAA gels and 3D culture within collagen gels. Next, 20 and 50 nm polystyrene nanospheres were incubated with MCF-7 cells in 2D or 3D cultures. Cell morphology and uptake efficiency was observed by confocal microscopy. Our study demonstrates that substrate stiffness differentially regulated cell morphology as well as the cell uptake behavior of polystyrene nanospheres in MCF-7 cells under 2D or 3D culture conditions. 1 Key words: matrix stiffness; 2D/3D cell culture; cell