纳米颗粒对人食管癌EC-9706、EC-109细胞增殖和凋亡的影响  

Effects of three kinds of nanoparticles on proliferation and apoptosis of esophageal squamous carcinoma cells

作　　者：韩鹏黎[1] 孙蕾[1] 吕朋举[1] 龚芬芬[1] 马超[1] 陈国[1] 朱怡然[2] 夏天[1] 曹巍[1] 

机构地区：[1]郑州大学附属郑州中心医院转化医学中心,河南郑州450007 [2]华中师范大学生命科学学院,湖北武汉430000

出　　处：《中国药理学通报》2016年第6期789-794,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 31570899);河南省医学科技攻关计划项目(No 201403273)

摘　　要：目的研究氧化铜(CuO)、氧化锌(ZnO)、二氧化钛(TiO_2)纳米颗粒在体外对人食管鳞状癌细胞系EC-9706和EC-109增殖的影响及其部分作用机制。方法使用透射电镜(TEM)、动态光散射(DLS)鉴定纳米颗粒物的理化性质,然后将不同浓度的TiO_2、CuO、ZnO(5~80 mg·L^(-1))与体外培养的EC-9706和EC-109细胞孵育,四甲基偶氮唑盐(MTT)法检测细胞生长抑制率,流式细胞术(FCM)法检测细胞周期及凋亡率,Transwell小室法检测细胞侵袭能力,蛋白免疫印迹法检测细胞中Bcl-2和caspase-3蛋白表达情况。结果实验所用的纳米CuO、ZnO、TiO_2颗粒为球形,粒径分别为12、20.6、12 nm。在水中与细胞培养液中聚集成较大颗粒并带负电。随着纳米颗粒浓度的增加,纳米CuO和ZnO颗粒可剂量依赖性抑制食管鳞癌细胞的增殖,G_0/G_1期细胞的比率上调,G_2/M期的比率下降,并促进凋亡率增加,侵袭力下降;纳米颗粒处理48 h后,与对照组相比,活化caspase-3表达量明显升高,Bcl-2表达量明显下降(P<0.05)。纳米TiO_2颗粒对食管鳞癌细胞的增殖、凋亡和侵袭能力无明显的影响。结论与纳米TiO_2相比,一定浓度的CuO和ZnO纳米颗粒可有效抑制食管鳞癌细胞的生长,阻滞细胞周期于G_0/G_1期,并诱导其凋亡,这可能是其抑制食管癌细胞功能的作用机制之一。Aim To study the effects of CuO,ZnO and TiO_2 nanoparticles on the viability and metastatic potential of EC-9706 and EC-109 esophageal squamous carcinoma cell line in vitro. Methods Characteristics of CuO,ZnO and TiO_2 nanoparticles were detected using transmission electron microscope( TEM) and dynamic light scattering( DLS). EC-9706 and EC-109 cells were treated with different concentrations of CuO,ZnO and TiO_2( 5 ~ 80 mg·L^(-1)). The cell proliferation was analyzed by MTT assay. The cell cycle and apoptotic rates were determined by flow cytometry( FCM). The cell invasion was assayed in Transwell chambers. The expression of Bcl-2 and caspase-3 protein in cells was detected by Western blot method. Results CuO,ZnO and TiO_2 nanoparticles were spherical with primary particle size 12,20. 6,12 nm. The particles were agglomerated in water and cell culture medium with negative charge. CuO and ZnO nanoparticles induced decreases in EC-9706 and EC-109 cell viability dose-dependently. After exposed to increasing concentrations of CuO and ZnO nanoparticles,the cell cycle analysis revealed a decreasing proportion of cells in G_2/ M and S phase,and up-regulation of the cells in G_0/ G_1 phase. Apoptotic cells also increased along with decreased cell invasion upon CuO and ZnO treatment.Nanoparticles treatment after 48 h, the activated caspase-3 expression quantity increased significantly and the Bcl-2 expression quantity decreased obviously( P < 0. 05) compared with control group. TiO_2 nanoparticles had no obvious effect on the EC-9706 and EC-109 cell proliferation,cell cycle,apoptosis and invasion. Conclusion Compared with TiO_2,CuO and ZnO nanoparticles can inhibit EC-9706 and EC-109 cell viability and metastatic potential,the mechanism of action involves cell cycle arrest in G_0/ G_1 phase and apoptosis. These findings can help the development of nanoparticles as anti-cancer therapeutics for esophageal cancer.

关 键 词：食管癌 氧化铜纳米颗粒 氧化锌纳米颗粒 二氧化钛纳米颗粒 细胞增殖 凋亡 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]