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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 effect of cisplatin (DDP) on HepG2 cells. Methods: The experiment was divided into GNPs, DDP, GNPs + DDP and no drug control group. The HepG2 cells were treated with the above groups of drugs, the proliferation inhibition rate of each group was detected by MTT assay, the apoptosis rate and cell cycle distribution were detected by FCM, the content of platinum in each group was quantified by inductively coupled plasma atomic emission spectrometry, Observation of changes of cell surface ultrastructure with atomic force microscopy. Results: The proliferation inhibition rates of HepG2 cells after treated with GNPs (1.0 nmol / L) and DDP (0.5, 1.0 or 2.0 μg / mL) were significantly higher than those in control group (26.19 ± 0.87% vs 36.17 ± 1.33% vs 39.86 ± 1.38 )%] Were significantly higher than that of pure DDP (0.5, 1.0 or 2.0 μg / mL) [(20.82 ± 0.76)%, (23.76 ± 1.04)% and (25.66 ± 1.16)%] Significance (P <0.01). The apoptosis rate of GNPs (1.0nmol / L) and DDP (2.0μg / mL) group (GNPs + DDP group) [(22.43 ± 3.24)%] was significantly higher than that of DDP group (P <0.05). The percentage of cells in S phase (59.05 ± 6.02%) was significantly higher than that in pure DDP group (40.37 ± 2.83%) (P <0.01). The level of DDP in HepG2 cells in GNPs + DDP group [(212.21 ± 12.3) μg / L] was significantly higher than that in DDP group (108.67 ± 7.74 μg / L) (P <0.01). Atomic force microscopy showed that, compared with DDP group, HepG2 cells in GNPs + DDP group showed an increase in pore size, roughness and decrease in cell nuclear fullness. Conclusion: GNPs can increase the accumulation of DDP in HepG2 cells, arrest the cells in S phase, enhance the inhibitory effect of DDP on cell proliferation and induce apoptosis.