用于逆转卵巢癌耐药的HMSN复合载药系统的研究

发布时间：2018-04-01 21:20

  本文选题：卵巢癌　切入点：多药耐药　出处：《东南大学学报(医学版)》2017年02期

【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p H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p H值的降低而逐渐升高,具有显著的p H敏感性,实现了以HMSN所处环境的p H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(P0.05),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。
[Abstract]:Aim: to construct a mesoporous silica HMSN complex drug carrier system and to investigate the reversal effect of the system on drug resistance of ovarian cancer cell line SKOV-3/ADR.Methods: the carboxyl group was modified on the surface of HMSN. The ADR and fluorescence were encapsulated by mechanical agitation and electrostatic attraction. The system was characterized by Zeta potential and transmission electron microscopy.The encapsulation efficiency, drug loading capacity and drug release rate in different pH values were determined, and then divided into two groups: HMSN-COOH@ADR fluorescent NVP group and free ADR fluorescent NVP group.The intracellular accumulation of HMSN compound drug loading system at different time points was observed by fluorescence confocal method, and the inhibition rate of cells in the two groups after 24 h exposure was detected by MTT assay, and the apoptosis rate of the two groups was detected by flow cytometry.The entrapment efficiency of ADR was 455.The entrapment efficiency of fluorescent NVP was 30. The drug release rate of 1.4%.HMSN compound drug loading system increased with the decrease of pH value and had significant sensitivity to pH.The release of the drug was regulated by the pH value of the environment in which HMSN was located, and the non-specific release of the compound drug loading system was effectively reduced, and it was found that the HMSN compound drug carrier system acted on the cells for 1 hour after fluorescence confocal observation.The amount of intracellular fluorescence increased obviously, and the fluorescence intensity increased gradually with the prolongation of the action time.The inhibition rate and apoptosis rate of MTT cells in HMSN-COOHADR fluorescent NVP group were significantly higher than those in free ADR fluorescent NVP group, and the cytotoxic effect of HMSN-COOHADR fluorescent NVP group was obviously enhanced.Conclusion: the complex drug loading system of HMSN-COOH@ADR fluorescent NVP can effectively reverse the drug resistance of ovarian cancer cell line SKOV-3/ADR. The mechanism of reversing drug resistance is to carry the complex drug carrier system into the cell through non-specific endocytosis pathway.It evades the recognition and efflux of ABC transporter superfamily such as P-gp.
【作者单位】： 东南大学医学院;东南大学附属中大医院妇产科;
【分类号】：R737.31
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本文编号：1697347