双硫仑螯合铜对MCF-7和MCF-10A细胞超微结构和力学特性的影响

Effects of DSF/Cu on surface ultrastructures and mechanical properties of human breast cancer and normal breast epithelial cells

目的:基于原子力显微镜(atomic force microscope,AFM)在纳米级分辨率和皮牛级力学测量探讨双硫仑(disulfiram,DSF)螯合氯化铜复合物(DSF/Cu)对人乳腺癌MCF-7和正常乳腺上皮MCF-10A细胞膜超微结构及力学特性的影响。方法:采用流式细胞术比较DSF/Cu作用于2种细胞后凋亡率和细胞周期的改变。AFM探测细胞表面形貌、超微结构、细胞高度、宽度及粗糙度的变化。利用AFM纳米压痕技术检测DSF/Cu对2种细胞硬度(杨氏模量)的影响。结果:流式细胞术结果显示:DSF/Cu可显著诱导MCF-7细胞凋亡,且呈浓度依赖性;而对MCF-10A细胞影响不大。细胞周期检测显示DSF/Cu对2种细胞的细胞周期影响不同。AFM成像观察发现,400和800 nmol/L的DSF/Cu作用于MCF-7细胞6 h后,细胞形态明显皱缩,体积变小,表面变得平整,丝状伪足明显回缩、变短,有些变成了板状伪足,甚至完全消失。进一步定量分析发现细胞宽度变小,高度增高,均方根粗糙度和平均粗糙度均减小,且呈浓度依赖性。而对MCF-10A细胞则影响甚微。最后,单细胞水平生物力学检测显示:DSF/Cu作用于MCF-7和MCF-10A细胞6 h后,2种细胞的杨氏模量随药物浓度的增加均有所增加,且呈浓度依赖性,但MCF-7细胞增加的比例要远远高于MCF-10A细胞。结论:DSF/Cu可能通过特异性影响MCF-7细胞的生物力学特性发挥高效低毒的抗乳腺肿瘤作用。

AIM： To study the effects of disulfiram/copper complex（ DSF/Cu） on ultrastructures and mechanical properties of human breast cancer and normal breast epithelial cells by atomic force microscopy（ AFM） based on the nanoscale resolution and piconewton force measurement level. METHODS： The change of cell cycle and apoptotic rate of MCF-7 cells and MCF-10 A cells induced by DSF / Cu were compared by flow cytometry. The cell surface morphology,ultrastructure,height,width and roughness were detected by AFM. The effects of DSF / Cu on the hardness（ Young＇s modulus） of the 2 kinds of cells were determined by AFM with indentation technique. RESULTS： DSF / Cu significantly induced apoptosis of the MCF-7 cells in a dose-dependent manner,whereas had little effect on the MCF-10 A cells. The cell cycle analysis showed that DSF / Cu induced G2/ M arrest in the MCF-7 cells,but led to G0/ G1 arrest in the MCF-10 A cells. The AFM images showed that the MCF-7 cells shrank and showed smaller and smoother morphology,and the filopodia were retracted obviously,even some became into lamellipodia,or disappeared completely after treated with DSF / Cu at concentrations of 400 and 800 nmol / L. The quantitative analysis indicated that the MCF-7 cells showed smaller width andlarger height,and the root mean square roughness and average roughness were decreased significantly in a dose-dependent manner after treated with DSF / Cu at concentrations of 400 and 800 nmol / L. However,little effect in the MCF-10 A cells was observed. The biomechanics test at a single cell level demonstrated that the Young＇s modulus of the MCF-7 cells and MCF-10 A cells were both increased,yet the proportion increased in the MCF-7 cells was much higher than that in the MCF-10 A cells after treated with DSF / Cu at concentrations of 400 and 800 nmol / L. CONCLUSION： DSF / Cu has strong antitumor effect on breast cancer with high efficiency and low toxicity by changing the properties of the biomechanics specifically.