纳米二氧化硅颗粒对HL-7702细胞黏附和迁移能力的影响  

Effects of SiO_2 nanoparticles on cell adhesion and migration of HL-7702 cells

作　　者：潘涛[1,2] 郭彩霞[3] 金明华[1] 刘晓梅[1] 刘颖[1] 杜海英[1] 孙志伟[1,4] 

机构地区：[1]吉林大学公共卫生学院卫生毒理学教研室,吉林长春130021 [2]西安医学院公共卫生系,陕西西安710021 [3]首都医科大学公共卫生学院劳动卫生与环境卫生学系,北京100069 [4]首都医科大学公共卫生学院卫生化学与毒理学系,北京100069

出　　处：《吉林大学学报（医学版）》2015年第4期732-736,共5页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81172704)

摘　　要：目的:探讨纳米二氧化硅颗粒对HL-7702细胞黏附迁移能力的影响,为其安全性评价提供实验依据。方法:体外培养的人正常肝细胞株HL-7702随机分为对照组和纳米二氧化硅颗粒暴露组,暴露浓度分别为12.5、25.0、50.0和100.0mg·L-1。采用透射电子显微镜(TEM)及动态光散射粒度分析仪(DLS)对纳米二氧化硅颗粒进行观察。细胞处理24h后,细胞黏附实验检测细胞黏附能力;划痕修复实验检测细胞迁移能力,TEM观察细胞对纳米二氧化硅颗粒的摄取及颗粒在细胞内的分布。结果:TEM观察,纳米二氧化硅呈圆形,颗粒大小均匀一致,分散性良好,颗粒平均粒径为(67.42±5.69)nm。DLS法检测,纳米二氧化硅颗粒在无血清RPMI-1640培养液中水合粒径为(134.13±2.78)nm,而在含1%、5%和10%血清的RPMI-1640培养液中的水合粒径明显增大。细胞黏附实验及划痕修复实验,与对照组比较,在无毒剂量下,纳米二氧化硅颗粒暴露组细胞的相对黏附率和损伤修复率明显下调(P<0.05),且随着作用浓度的升高下调作用愈加明显。与对照组比较,在无毒剂量作用下,纳米二氧化硅暴露组大量的纳米二氧化硅颗粒已被细胞摄取,颗粒主要以成簇的形式分布于内吞小泡中或散在分布于胞质中,可见细胞膜凹陷、包裹吞噬纳米颗粒的过程。结论:纳米二氧化硅颗粒能够抑制HL-7702细胞的黏附和迁移能力,并呈一定的浓度依赖效应。Objective To study the effects of SiO2 nanoparticles（SiO2 NPs）on the cell adhesion and migration of HL-7702 cells,and to provide experimental basis for safety evaluation of SiO2 NPs.Methods The in vitro cultured human normal liver cell line HL-7702 were randomly divided into control and SiO2 NPs exposure groups at the concentrations of 12.5,25.0,50.0,and 100.0 mg· L^-1,respectively.Transmission electron microscope（TEM）and dynamic light scattering（DLS）method were used to observe the characteristics of SiO2 NPs.After24 hexposure,cell adhesion assay was applied to examine the changes of cell adherence;wound healing assay was used to discover the changes of cell migration;the cellular uptake and biodistribution of SiO2 NPs were observed using TEM.Results Based on the result of TEM,SiO2 NPs were spherically shaped,uniformly sized and sporadically dispersed with the diameter of（67.42±5.69）nm.The results of DLS method showed that the average hydrated particle size of SiO2 NPs dispersing in RPMI-1640serum-free medium was（134.13±2.78）nm,while the hydrated particle sizes of SiO2 NPs dispersing in RPMI-1640 with 1%,5%,or 10% serum were apparently enlarged.The results of cell adhesion and wound healing assays showed that the adhesion rates and the healing rates of cells could be down-regulated by SiO2 NPs even under a dose that did not affect the cell viability and membrane integrity（P〈0.05）,furthermore,the down regulation effect was significantiy increased along with the enlargement of the SiO2 NPs＇ concentration.The TEM results showed that after exposed to nontoxic dose of SiO2 NPs,the cells accumulated a large number of nanoparticles,the particles mostly dispersed in endocytic recycling compartment,and some were in the cytoplasm;some parts of the cell membrans were caved,and the procedure of endocytose could be captured.Conclusion SiO2 NPs could inhibit the cell adhesion and migration in a dosedependent manner.

关 键 词：纳米二氧化硅颗粒 HL-7702细胞 细胞黏附 细胞迁移 细胞摄取 

分 类 号：R992[医药卫生—毒理学]