纳米金增强顺铂对肝癌HepG2细胞的细胞毒作用  

Gold nanoparticles enhance the cytotoxic effects of cisplatin on liver cancer HepG2 cells

作　　者：巫青[1] 潘运龙[1] 覃莉[2] 赵晓旭[1] 丁晖[1] 皮江[3] 傅岳武[1] 胡杨志[1] 蔡继业[3] 

机构地区：[1]暨南大学附属第一医院普外科,广东广州510630 [2]暨南大学医学院组织胚胎学教研室,广东广州510632 [3]暨南大学生命科学技术学院纳米生物技术实验室,广东广州510632

出　　处：《肿瘤》2012年第9期675-680,共6页Tumor

基　　金：国家自然科学基金资助项目(编号:30772131);国家重点基础研究发展计划(973计划)资助项目(编号:2010CB833603);2008年广东省自然科学基金面上项目[编号:粤科基办字(2008)4号];2012年暨南大学第一临床医学院科研培育专项基金项目(编号:No.4)

摘　　要：目的:观察纳米金(gold nanoparticles,GNPs)是否能够增强顺铂(cisplatin,DDP)对肝癌HepG2细胞的细胞毒作用。方法:实验分成GNPs、DDP、GNPs+DDP和不加药对照组。采用上述各组药物处理HepG2细胞后,MTT法检测各组细胞的增殖抑制率,FCM检测细胞凋亡率和细胞周期分布情况,电感耦合等离子体原子发射光谱仪定量检测各组细胞内的铂含量,原子力显微镜下观察细胞表面超微结构的变化。结果:GNPs(1.0nmol/L)与DDP(0.5、1.0或2.0μg/mL)联合作用后,HepG2细胞的增殖抑制率[(26.19±0.87)%、(36.17±1.33)%和(39.86±1.38)%]均明显高于相应浓度的单纯DDP(0.5、1.0或2.0μg/mL)组[(20.82±0.76)%、(23.76±1.04)%和(25.66±1.16)%],差异有统计学意义(P<0.01)。GNPs(1.0nmol/L)联合DDP(2.0μg/mL)组(GNPs+DDP组)的细胞凋亡率[(22.43±3.24)%]明显高于单纯DDP(2.0μg/mL)组(DDP组)[(14.17±2.48)%](P<0.05),S期细胞百分比[(59.05±6.02)%]也较单纯DDP(2.0μg/mL)组[(40.37±2.83)%]明显增加(P<0.01)。GNPs+DDP组HepG2细胞内的DDP含量[(212.21±12.3)μg/L]较DDP组[(108.67±7.74)μg/L]明显增加(P<0.01)。原子力显微镜下观察发现GNPs+DDP组HepG2细胞与DDP组相比,细胞膜表面孔径增大、粗糙度增加,细胞核饱满程度下降。结论:GNPs可增加DDP在HepG2细胞内的积聚,使细胞阻滞于S期,增强DDP对细胞增殖的抑制作用,诱导细胞凋亡。Objective： To observe whether gold nanoparticles （GNPs） can enhance the cytotoxic effects of cisplatin （DDP） on liver cancer HepG2 cells. Methods： The HepG2 cells were treated with GNPs alone, DDP alone or GNPs combined with DDP, and the cells without any treatment served as a control. The inhibitory rate of HepG2 cell proliferation was detected by MTT method. The apoptotic rate and the cell cycle distribution were examined by flow cytometry （FCM）. The concentration of DDP in HepG2 cells was determined by inductive coupled plasma atomic emission spectrometer （ICP-AES）. The ultrastructural change of cell surface was observed under an atomic force microscope （AFM）. Results： The inhibitory rate of HepG2 cell proliferation in GNPs （1.0 nmol/L） combined with DDP （0.5, 1.0 or 2.0 μg/mL） group [（26.19±0.87）%, （36.17±1.33）% and （39.86±1.38）%] was higher than those in DDP （0.5, 1.0 and 2.0 μg/mL） alone groups [（20.82±0.76）%, （23.76±1.04）% and （25.66±1.16）%] （P 〈 0.01）. The apoptotic rate of GNPs （1.0 nmol/L） combined with DDP （2.0 μg/mL） group （GNPs ＋ DDP group） [（22.43±3.24）%] was higher than that of DDP （2.0 μg/mL） alone group （DDP group） [（14.17±2.48）%] （P 〈 0.05）, and the percentage of HepG2 cells arrested in S phase was higher in GNPs ＋ DDP group [（59.05±6.02）%] as compared with the DDP group [（40.37±2.83）%] （P 〈 0.01）. The intracellular concentration of DDP was also higher in GNPs ＋ DDP group [（212.21±12.3） μg/L] as compared with the DDP group [（108.67±7.74） μg/L] （P 〈 0.01）. AFM image showed the HepG2 cells having larger pore size of cell membrane, more rough cell surface, and a shrink in the size of nucleus in GNPs ＋ DDP group as compared with the DDP group. Conclusion： GNPs can increase the accumulation of DDP in HepG2 cells, and arrests the cells in S phase, which results in an increase in the inhibitory effect of DDP on cell proliferation and induces apopt

关 键 词：肝肿瘤 肿瘤治疗方案 顺铂 纳米金 细胞 HepG2 

分 类 号：R735.7[医药卫生—肿瘤]