阿霉素纳米粒对白血病耐药细胞株K562/DOX耐药逆转作用的研究  

Reversing Effect of Doxorubicin-loaded Nanoparticles on Multidrug Resistant Cell Line K562/DOX

作　　者：王华芳[1] 胡豫[1] 郑金娥[1] 周薇[1] 方峻[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液病研究所,武汉430022

出　　处：《华中科技大学学报（医学版）》2004年第3期289-292,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目 (No 30 170 392 )

摘　　要：目的　合成阿霉素聚氰基丙烯酸异丁酯纳米粒 ,观察其对阿霉素耐受性细胞株K5 6 2 (K5 6 2 /DOX)耐药性逆转效果并探讨可能机制。方法　采用乳液聚合法合成纳米粒 ,透射电镜观察计算粒径 ,分光光度法计算包封率 ;以梯度浓度的阿霉素纳米粒处理培养的K5 6 2 /DOX细胞 ,噻唑蓝比色法 (MTT)检测K5 6 2 /DOX细胞的半数生长抑制浓度 (IC50 )并计算逆转倍数 ;流式细胞术检测细胞内药物浓度 ;TUNEL法检测药物诱导后凋亡的发生及凋亡率。结果　阿霉素纳米粒平均粒径 (98 5± 3 7)nm ,包封率为 95 % ;MTT结果显示阿霉素纳米粒对K5 6 2 /DOX细胞的生长抑制率较单纯阿霉素显著增高 ,阿霉素纳米粒对K5 6 2 /DOX的耐药逆转倍数为 6 2 ;TUNEL检测经 10 μg/ml阿霉素纳米粒处理后的K5 6 2 /DOX平均凋亡率为 38 7%。结论　以聚氰基丙烯酸异丁酯纳米粒为载体的阿霉素可增强诱导K5 6 2 /DOX细胞凋亡的效率和有效逆转其耐药性。Objective To investigate the reversing effect of doxorubicin-loaded polyisobultylcyanoacrylate nanoparticles (NP-DOX) on multidrug resistant cell line K562/DOX and the possible mechanism.Methods Polyisobultylcyanoacrylate nanoparticles were prepared by emulsion polymerization. The size of nanoparticles was determined by electron transmission microphotography and the percentage of doxorubicin associated with the nanoparticles was determined by spectrophotometer. Multidrug resistant cell line K562/DOX was established and incubated with series concentrations of doxorubicin-loaded nanoparticles. The growth inhibitory ratio and IC 50 were calculated by MTT colorimetry assay. The average drug concentration in K562/DOX cells was evaluated by flow cytometry. Apoptosis and apoptotic ratio following induction by doxorubicin-loaded nanoparticles were observed by TUNEL staining. Results The average diameter of doxorubicin-loaded nanoparticles was (98.5±3.7) nm and the percentage of doxorubicin associated with the nanoparticles was 95 %. MTT revealed that the growth inhibitory ratio of K562/DOX cells treated by NP-DOX was significantly higher than that by free DOX, and the IC 50 of DOX was 6.2 times than that of NP-DOX. TUNEL demonstrated that the average apoptotic ratio of K562/DOX cells treated with 10 μg/ml NP-DOX was 38.7 %. Conclusion NP-DOX was more effective to induce apoptosis in K562/DOX cells than DOX. Doxorubicin-loaded nanoparticles can reverse multidrug resistance of K562/DOX cells by elevating the drug concentration significantly.

关 键 词：阿霉素 K562 纳米粒 凋亡率 耐药逆转作用 耐药细胞株 白血病 丁酯 丙烯酸 合成 

分 类 号：R733[医药卫生—肿瘤] R737[医药卫生—临床医学]