ATP7B siRNA阳离子纳米脂质体逆转卵巢癌细胞株顺铂耐药的体内外实验研究  

The Reversal of Cisplatin Resistance in an Ovarian Cancer Cell Line by Cationic Liposomes Loaded with ATP7B Small Interfering RNA

作　　者：钱艺美[1] 王军青[1] 周闺臣[2] 陈菡[2] 

机构地区：[1]解放军第101医院,江苏无锡214000 [2]第二军医大学药学院,上海200433

出　　处：《实用妇产科杂志》2012年第12期1030-1034,共5页Journal of Practical Obstetrics and Gynecology

摘　　要：目的:探讨负载P型铜转运ATP酶(ATP7B)、小干扰RNA(siRNA)阳离子纳米脂质体(ATP7B/DC-Lip)逆转卵巢癌细胞株SKOV3顺铂(DDP)耐药的作用及意义。方法:通过DDP梯度诱导法构建卵巢癌耐药细胞株SKOV3/DDP,利用细胞增殖抑制实验测定其耐药指数。薄膜分散法制备ATP7B/DC-Lip并测定其各项纳米表征。利用细胞增殖抑制实验和裸鼠体内实验分别检测ATP7B/DC-Lip的体内外抗肿瘤效果。结果:成功构建了卵巢癌耐药细胞株SKOV3/DDP,ATP7B/DC-Lip在体外条件下可以分别沉默75%(48小时)和78%(72小时)的ATP7B蛋白表达。在体内外抑制肿瘤生长实验的结果显示,当转染ATP7B/DC-Lip后,体外条件下SK-OV3/DDP的IC50显著下降为3.51±0.93μg/ml(n=6);体内DDP耐药型肿瘤模型中DDP+ATP7B/DC-Lip组的肿瘤大小较其他组明显更小(P<0.05)。结论:ATP7B/DC-Lip可以逆转卵巢癌细胞株SKOV3的DDP耐药,可能作为一种有效的纳米药物用于治疗DDP耐药性卵巢癌。Objective： The aim of this study was to investigate the effect and mechanism of the reversal of Cisplatin （DDP） resistance in an ovarian cancer cell line I^y cationic liposomes loaded with ATP7B small in terfering RNA （siRNA） （ATP7B/DC-Lip）. Methods：A SKOV3 ovarian cancer cell line which is resistant to DDP （SKOV3/DDP） was developed by a gradient induction method with DDP. Cell proliferation assays were used to determine the resistance index of SKOV3/DDP. Furthermore, ATP7B/DC-Lip was developed by a thin-film method,and the size and encapsulation efficiency of siRNA of ATP7B/DC-Lip were measured. Cell proliferation assays and in vivo antitumor assays were performed to evaluate the in vitro and in vivo antitumor activity of reagents. Resulte：SKOV3/DDP was successfully developed by a gradient induction method with DDP,ATP7B/DC-Lip could significantly silence the protein expression of ATP7B. The cell proliferation assay showed that ATP7B/DC-Lip significantly reversed the DDP resistance of SKOV3/DDP,with a much reduced IC50 of 3. 51 ± 0. 93 pg/ml （ n = 6）. Furthermore,ATP7B/DC-Lip combined with DDP significantly inhibited the tumor development and possessed the best antitumor activity compared with other three groups （nega- tive controI,ATP7B/DC-Lip and DDP） （ P 〈 0.05）. Conclusions： ATPTB/DC-Lip could significantly reverse the DDP resistance of SKOV3/DDP both in vitro and in vivo, suggesting that it may serve as an effective nanomedicine in treating ovarian cancer with DDP resistance.

关 键 词：卵巢癌 耐药 肿瘤治疗 P型铜转运ATP酶 小干扰RNA 

分 类 号：R737.31[医药卫生—肿瘤]