吞噬二氧化硅微粒对HepG2细胞黏附和迁移的影响  

EFFECT OF CELLULAR UPTAKE OF SiO_2 PARTICLES ON ADHESION AND MIGRATION OF HepG2 CELLS

作　　者：张裕英[1] 胡玲[1] 高长有[1] 

机构地区：[1]高分子合成与功能构造教育部重点实验室浙江大学高分子科学与工程学系,杭州310027

出　　处：《高分子学报》2009年第8期815-822,共8页Acta Polymerica Sinica

基　　金：国家自然科学基金(基金号50873087;50425311);国家重点基础研究发展规划项目(973计划;项目号2005CB623902)资助

摘　　要：通过Stber溶胶-凝胶法制备了掺杂荧光染料的二氧化硅微粒.透射电子显微镜表征其直径分别为80、160和500nm.在荧光显微镜下观察HepG2细胞对不同尺寸微粒的吞噬并采用流式细胞仪研究了微粒进入细胞的途径.检测了二氧化硅微粒的细胞毒性,通过划痕修复实验、细胞黏附和Transwell细胞迁移实验研究了吞噬二氧化硅微粒对细胞黏附和迁移能力的影响.实验结果表明,HepG2细胞主要通过网格蛋白介导的胞吞途径对二氧化硅微粒进行吞噬,4℃培养和叠氮化钠处理都会抑制胞吞的效率.在浓度为0～200μg/mL范围内,直径为80nm的二氧化硅微粒会对细胞造成浓度依赖的细胞毒性,而直径为160nm和500nm的二氧化硅微粒没有对细胞存活率造成明显的影响.但是,吞噬三种尺寸的微粒后,细胞的黏附和迁移能力都有较明显的下降,推测原因可能是由于胞吞过程对细胞骨架造成了损伤.The dye-doped SiO2 particles with diameters of 80,160 and 500 nm were prepared by using the Stoeber sol-gel method. Cellular uptake of the luminescent SiO2 particles by HepG2 cells and the endocytic pathway were studied by fluorescent microscopy and flow cytometry. Influence of the SiO2 particles on HepG2 cells was tested in terms of cytotoxicity, cell adhesion and migration by MTT assay, cell adhesion assay, the wound healing and transwell assay. According to the results of flow cytometry, the uptake efficiency of the SiO2 particles was significantly decreased when the culture was performed at 4℃ as well as in the presence of sodium azide or amandatine-HCl, while only very limited reduction was observed with the addition of genestein, revealing that the clathrin-mediated endocytosis was the main uptake pathway. The MTF a dose-dependent manner, while the 160 nm and assay found that the 80 nm SiO2 particles caused cytotoxicity in 500 nm particles didn＇t show significant cytotoxicity in a concentration range of 0 - 200 μg/mL. However, the results of the wound healing assay, cell adhesion and transwell assay demonstrate that the cell adhesion and migration were greatly inhibited by uptake of the SiO2 particles regardless of their size, most possibly because the endocytic process leads to disruption of the cytoskeleton. These studies suggest that the effect of endocytosis on functions of cells should also be assessed to evaluate the biocompatibility of nanoparticles.

关 键 词：二氧化硅 胞吞 细胞毒性 细胞迁移 

分 类 号：R114[医药卫生—卫生毒理学]