肺腺癌细胞对谷胱甘肽纳米Fe_3O_4的内吞作用

Endocytosis of lung adenocarcinoma cell on glutathione modified magnetite(Fe_3O_4)nanoparticles

目的在体外观察人肺腺癌细胞(SPC-A1)对谷胱甘肽(GSSG)修饰的纳米Fe3O4的内吞作用及与培养温度、时间及浓度的关系。方法通过透射电镜(TEM)与普鲁士兰染色法观察SPC-A1细胞对GSSG修饰的纳米Fe3O4的内吞作用。采用原子吸收光谱(AAS)法进行SPC-A1细胞粘附量及内吞量的定量分析。结果TEM结果表明,在4℃下培养24 h后GSSG修饰的纳米Fe3O4吸附于SPC-A1细胞表面;37℃下培养1 h后SPC-A 1细胞可内吞GSSG修饰的纳米Fe3O4细胞数目明显增加。AAS分析表明,SPC-A1细胞表面纳米Fe3O4的粘附量同培养时间及浓度呈正相关。GSSG修饰的纳米Fe3O4浓度较低(0.1或0.4 mg/ml)时,内吞量在24 h后达到饱和;GSSG修饰的纳米Fe3O4浓度较高(0.7或1.0 mg/ml)时,内吞量在72 h后达到饱和,单个细胞内吞量最大可达160 pg。结论SPC-A1细胞对GSSG修饰的纳米Fe3O4的内吞需要能量,内吞量依赖于浓度及培养时间。

Objective To study the relationship between endoeytosis of human lung adenocarcinoma cell line SPC- A1 modified by oxidized Glutathione（GSSG） and culture conditions including culture temperature, time and concentration. Methods The uptake of GSSG- modified Fe3O4 nanopartieles by SPC- A1 cell was observed by transmission electron microscopy （TEM） and Prussian blue staining. The extracellular adhesion or intracellular uptake of GSSC- - modified Fe3O4 nanopartieles were measured by atom absorb spectrum （AAS）. Results TEM results indicated that GSSG - modified Fe3O4 nanopartieles adhered on the membrane of SPC - A1 cell after being cultured at 4 ℃ for 1 h or 24 h, but SPC - A1 cell could took up nanopartieles particles after cultured at 37 ℃ for 1 h. Prussian blue staining indicated that SPC - A1 cells containing GSSG - modified Fe3O4 nanopartieles were much more with the prolongation of culture time or the increase of concentration. AAS analysis indicated that the extraeellular adhesion of GSSG- modified magnetic particles was depended on culture time and concentration. The uptake amount of GSSG- modiied Fe3O4 nanopartieles by SPC - A1 cell was saturated after 24 h when the nanopartieles in low concentration （0.1 mg/ml or 0.4 mg/ml）, the uptake was saturated after 72 h when the nanoparti- eles in high concentration （0.7 mg/ml or 1.0 mg/ml）. The maximal amount per SPC - A1 cell could reach 160 pg. Conclusion The uptake of GSSG - modified Fe3O4 nanoparticles by SPC - A1 cell is energy needed. It depends on the concentration of nanoparticles and saturates at certain time.