用于逆转卵巢癌耐药的HMSN复合载药系统的研究-东南大学学报医学版

用于逆转卵巢癌耐药的HMSN复合载药系统的研究

单位：1. 东南大学医学院, 江苏南京 210009;
2. 东南大学附属中大医院妇产科, 江苏南京 210009

分类号：R737.31

出版年·卷·期（页码）：2017·36·第二期（148-154）

摘要：

目的：构建介孔二氧化硅（HMSN）复合载药系统，探究该系统对卵巢癌耐药细胞株SKOV-3/ADR耐药的逆转作用。方法：在HMSN表面修饰羧基，通过机械搅拌和静电吸引的作用包载ADR和荧光NVP；采用Zeta电位、透射电镜等方法对此系统进行表征，并测定此系统的包封率和载药量以及在不同pH值环境中的药物释放率；再将实验分为两组，即HMSN-COOH@ADR荧光NVP组和游离ADR荧光NVP组，采用荧光共聚焦观察不同时间点HMSN复合载药系统在胞内的蓄积情况；利用MTT法检测细胞在两组作用24 h后的抑制率，同时通过流式细胞分析仪检测两组的凋亡率。结果：Zeta电位表明HMSN在修饰羧基之前其所带电荷约为-20 mV，在修饰羧基后HMSN所带电荷增加至约-40 mV；通过透射电镜表明此载药系统在包载药物前后的形态和粒径均未发生明显改变；测定HMSN复合载药系统中ADR的包封率为45%，载药量为7.5%，荧光NVP的包封率为30%，载药量为1.4%。HMSN复合载药系统的药物释放率随着环境pH值的降低而逐渐升高，具有显著的pH敏感性，实现了以HMSN所处环境的pH值为“开关”调控药物的释放，有效地减少了复合载药系统在胞外的非特异性释放；通过荧光共聚焦观察发现HMSN复合载药系统作用细胞1 h后，其在胞内的蓄积量即可明显增加，随着作用时间的延长胞内的荧光强度也在逐渐增强；药物作用相同的时间，HMSN-COOH@ADR荧光NVP组较游离ADR荧光NVP组的MTT细胞抑制率和细胞凋亡率均明显提高（P<0.05），对细胞的毒性作用明显增强。结论：构建的HMSN-COOH@ADR荧光NVP的复合载药系统可以有效逆转卵巢癌耐药细胞株SKOV-3/ADR的耐药性，其逆转耐药的机制是通过非特异性的内吞途径携带复合载药系统进入细胞内，逃避了P-gp等ABC运载蛋白超家族的识别和外排作用。

Objective: To develop a co-delivery system based on hollow mesoporous silica nanoparticles(HMSN), and explore the effect of this system in reversing multiple drug resistance in ovarian cancer cell SKOV-3/ADR. Methods: HMSN was selected as the nanocarriers, with-COOH modified on the surface and ADR, NVP-AEW541 loaded inside. Then this system was characterized by Zeta potential, TEM and other methods, and the entrapment efficiency, drug loading rate, drug release rate in different pH environment were detected; Experiment was carried out in two groups, HMSN-COOH@ADR fluorescent NVP and free ADR fluorescent NVP group, gathering of the co-delivery system in cells and nucleus was observed by LSCM at different time points; Cells apoptosis and MTT cell inhibition rate were also detected after 24 hours in each group. Results: Zeta potential showed that the charge of HMSN was about-20 mV before modified carboxyl, and rised to-40 mV after modified carboxyl; The size and shape of HMSN were not changed after loading drugs by TEM; The entrapment efficiency was 45% of ADR and 30% of fluorescent NVP, drug loading rate was 7.5% of ADR and 1.4% of fluorescent NVP; The drugs release rate could be significantly improved as the pH value decreased, so the pH value was used as a "switch" which could effectively reduce extracellular nonspecific drug release. The intracellular quantity of HMSN drug delivery system was increased after one hour, and enhanced gradually with time extension. After the same time, the cell apoptosis rate and MTT inhibition rate of HMSN-COOH@ADR fluorescence NVP group were both higher than that of free ADR fluorescent group (P<0.05), indicated the increased cytotoxicity on cells. Conclusion: This HMSN drug delivery system can enter into cells mainly through nonspecific endocytosis, effectively reverse multidrug resistance of ovarian cancer cell SKOV-3/ADR,thus avoide the identification and efflux effect of P-gp.

参考文献：

[1] CRAVEIRO V,YANG-HARTWICH Y,HOLMBERG J C,et al.Phenotypic modifications in ovarian cancer stem cells following Paclitaxel treatment[J].Cancer Med,2013,2(6):751-762.
[2] TOMONO T,KAJITA M,YANO K,et al.,Adenovirus vector infection of non-small-cell lung cancer cells is a trigger for multi-drug resistance mediated by P-glycoprotein[J].Biochem Biophys Res Commun,2016,476(4):183-187.
[3] WU Q,YANG Z,NIE Y,et al.,Multi-drug resistance in cancer chemotherapeutics:mechanisms and lab approaches[J]. Cancer Lett,2014,347(2):159-166.
[4] LI W,ZHANG H,ASSARAF Y G,et al.Overcoming ABC transporter-mediated multidrug resistance:molecular mechanisms and novel therapeutic drug strategies[J].Drug Resistance Updates,2016,27:14-29.
[5] JUN Y W,LEE J HCHEON J,Chemical design of nanoparticle probes for high-performance magnetic resonance imaging[J].Angew Chem Int Ed Engl,2008,47(28):5122-5135.
[6] TANG C,RUSSELL P J,MARTINIELLO-WILKS R,et al.,Concise review:Nanoparticles and cellular carriers-allies in cancer imaging and cellular gene therapy?[J].Stem Cells,2010,28(9):1686-1702.
[7] 蔡阳,朱传东,郑勤.金纳米在肿瘤治疗中的应用[J].现代医学,2016,44(6):893-896.
[8] DOUGLAS J B,SILVERMAN D T,POLLAK M N,et al.Serum IGF-I,IGF-II,IGFBP-3,and IGF-I/IGFBP-3 molar ratio and risk of pancreatic cancer in the prostate,lung,colorectal,and ovarian cancer screening trial[J].Cancer Epidemiol Biomarkers Prev,2010,19(9):2298-2306.
[9] JIN M,BUCK EMULVIHILL M J.Modulation of insulin-like growth factor-1 receptor and its signaling network for the treatment of cancer:current status and future perspectives[J].Oncol Rev,2013,7(1):e3.
[10] PANYAM J,LABHASETWAR V.Biodegradable nanoparticles for drug and gene delivery to cells and tissue[J].Advanced Drug Delivery Reviews,2012,64:61-71.
[11] LEE E S,GAO ZBAE Y H.Recent progress in tumor pH targeting nanotechnology[J].J Control Release,2008,132(3):164-170.
[12] HUANG Y,JIANG Y,WANG H,et al.Curb challenges of the "Trojan Horse" approach:smart strategies in achieving effective yet safe cell-penetrating peptide-based drug delivery[J].Adv Drug Deliv Rev,2013,65(10):1299-1315.
[13] VANAMALA J,REDDIVARI L,RADHAKRISHNAN S,et al.Resveratrol suppresses IGF-1 induced human colon cancer cell proliferation and elevates apoptosis via suppression of IGF-1R/Wnt and activation of p53 signaling pathways[J].BMC Cancer,2010,10:238.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录