碳纳米管携载多柔比星对人卵巢癌耐药细胞体外抑制研究  

Inhibition of adriamycin carried by single-walled carbon nanotubes on human ovarian carcinoma SKOV3 adriamycin-resistant cells in vitro

作　　者：刘思思[1] 黎雁英[2] 裴蕾[1] 田佳[1] 林静[1] 周英琼[1] 

机构地区：[1]桂林医学院附属医院病理科,广西桂林541001 [2]桂林医学院第二附属医院内科,广西桂林541001

出　　处：《中华肿瘤防治杂志》2016年第14期924-929,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：桂林市人口健康安全技术开发与应用(20140120-1-11)

摘　　要：目的长期应用多柔比星(doxorubicin,ADM)能够增加药物耐受性及毒副作用,单壁碳纳米管(single-walled carbon nanotubes,SWCNTs)作为药物载体可提高载药率。本研究通过制备携载ADM的碳纳米管复合物然后作用于人卵巢癌ADM耐药细胞株,探讨研究碳纳米管复合物对细胞的杀伤作用。方法制备ADM/PEG/SWCNTs复合物,傅里叶变换红外光谱扫描仪表征检测PEG/SWCNTs表面官能团的变化;紫外分光光度计测定载药量;WST-1法检测ADM/PEG/SWCNTs对SKOV3/ADM的增殖抑制作用;流式细胞术和Hoechst33258荧光染色观察ADM/PEG/SWCNTs对细胞凋亡的影响;PCR及蛋白质印迹法检测ADM/PEG/SWCNTs对MDR-1 mRNA及P-gp糖蛋白的表达影响。结果成功制备ADM/PEG/SWCNTs复合药物,其中ADM在PEG/SWCNTs的载药率为(79.5±10.6)%,包封率(32.0±4.2)%。WST-1实验显示,ADM/PEG/SWCNTs及ADM作用细胞平均IC50为(18.0±1.2)和(37.5±1.9)μg/mL。ADM/PEG/SWCNTs对SKOV3/ADM细胞24、48和72h细胞增殖抑制率分别为(43.27±3.60)%、(53.40±8.12)%和(68.00±4.26)%,组间比较,差异有统计学意义,P<0.001。流式细胞术检测ADM/PEG/SWCNTs组及ADM组SKOV3/ADM细胞凋亡率为(14.83±1.86)%和(5.28±0.45)%,F=137.292,P=0.001。Hoechst33258染色ADM/PEG/SWCNTs组细胞呈明显凋亡形态改变。ADM/PEG/SWCNTs组可下调MDR-1mRNA及P-gp蛋白的表达,经灰度扫描分析,组间比较,差异有统计学意义,F=18.634,P=0.003。结论体外制备的ADM/PEG/SWCNTs纳米复合物具有高载药率,可抑制人卵巢癌细胞多柔比星耐药细胞增殖,促进人卵巢癌ADM耐药细胞凋亡,可下调MDR-1mRNA及P-gp蛋白的表达。OBJECTIVE Long time application of adriamycin could incerease the tolerance and side effects,as sigle- walled carbon nanotubes, drug carrier, could increase the drug loading rate. This paper aimed to study the anticancer poten- tial of single-walled carbon nanotubes loaded with Adriamycin and Polyethylene glycol in human ovarian carcinoma adria- mycin-resistant cells. METHODS ADM/PEG/SWCNTs composites were prepared. FTIR was used to characterize the change of surface functional groups of PEG/SWCNTs. The drug loading efficiencies of Adriamycin in the ADM/PEG/ SWCNTs composites were analyzed by infrared spectroscopy. The inhibitory effect of ADM/PEG/SWCNTs composites on proliferation of human SKOV3/ADM cells was evaluated by the WST-1 assay, and the apoptotic effect was demonstra- ted by flow cytometrys and Hoechst33258 fluorescence staining. RT-PCR and Western Blot were employed to measure the expressions of MDR-lmRNA and P-gp protein respectively. RESULTS ADM/PEG/SWCNTs composites were pre- pared successfully, and the drug loading rate of Adriamycin in the ADM/PEG/SWCNTs composites was （79. 5 ± 10. 6）%, and the encapsulation efficiency was （32.0±4.2）%. The half maximal inhibitory concentration of ADM/PEG/SWCNTs and ADM was （18.0±1. 2） μg/mL and （37.5±1.9） μg/mL. The inhibition rates of ADM/PEG/SWCNTs on SK- OV3/ADM cells for 24 h,48 h and 72 h were （43.27±3.60）%, （53.40±8.12% and （68.00±4.26）% ,respectively. There was a significant difference among these groups （P〈 0. 001 ）. Flow cytometry showed that ADM/PEG/SWCNTs group and ADM drug group apoptosis rates were （14.83±1.86）% vs （5.28±0. 45）% （F=137. 292, P=0. 001）. The SKOV3/ADM cells treated with ADM/PEG/SWCNTs showed typical morphological change of apoptosis by Hoechst33258 fluorescence stai- ning. ADM/PEG/SWCNTs could down regulate the expression of MDRdmRNA and P-gp protein. There was a significant difference among these groups （F=18. 634, P=0. 003）. CONCLUSION The ADM/PEG/SWCNTs composites have high

关 键 词：卵巢癌 多柔比星 耐药 单壁碳纳米管 

分 类 号：R737.31[医药卫生—肿瘤]