载紫杉醇的聚乙二醇聚丙交酯-乙交酯聚赖氨酸纳米粒制备及其对耐药卵巢癌细胞的作用

Preparation of paclitaxel-loaded mPEG-PLGA-PLL nanoparticles and their effects on drug-resistant ovarian cancer cells

目的制备聚乙二醇聚丙交酯-乙交酯聚赖氨酸(mPEG-PLGA-PLL)包载紫杉醇(PTX)的纳米粒(PTX NPs),探讨其在体内外对耐药卵巢癌细胞的作用。方法以包封率和粒径为综合评价指标,筛选PTX NPs的最佳制备方法,并采用正交实验优化该制备方法的工艺参数。通过粒度仪和透射电镜对PTX NPs进行表征分析。采用高效液相色谱法分析定量检测PTX的包封率,透析法评估PTX NPs中PTX的释放率。荧光倒置显微镜观察PTX NPs的摄取情况,CCK-8法检测NPs及PTX NPs对耐药卵巢癌细胞生长影响,采用动物成像仪观察PTX NPs在体内的生物学分布。结果PTX NPs粒径为(103.0±0.9)nm,包封率为(84.1±1.3)%,具有一定缓释作用。细胞摄取实验结果显示PTX NPs相比游离药物更容易被卵巢癌A2780细胞紫杉醇耐药株(A2780TR)细胞摄取。空载mPEG-PLGA-PLL纳米粒无明显细胞毒性,游离PTX在A2780和A2780TR细胞中的IC50分别为(0.0102±0.0003)μg·mL^-1和(4.55±0.44)μg·mL^-1,而PTX NPs的IC50分别为(0.0031±0.0006)μg·mL^-1和(1.05±0.13)μg·mL^-1。体内生物学分布结果显示PTX NPs能够靶向肿瘤部位发挥作用。结论本研究制备的PTX NPs粒径适宜、包封率高、具有药物缓释作用,易被细胞摄取,相比游离PTX对卵巢癌耐药细胞的作用更强,且具有体内靶向作用,在卵巢癌治疗中具有一定的应用前景。

AIM To prepare polyethylene glycol polylactide-glycolide polylysine(mPEG-PLGA-PLL)encapsulated paclitaxel(PTX)nanoparticles(PTX NPs),and to explore its effects on drug-resistant ovarian cancer cells in vivo and in vitro.METHODS In order to screen the best preparation method of PTX NPs,the orthogonal experiments were used to optimize the process parameters of the preparation method.The encapsulation efficiency and particle size were the comprehensive evaluation indexes.Zetasizer and the transmission electron microscope were used to determine the characteristics of PTX NPs.High performance liquid chromatography was used to detect the encapsulation rate of PTX in PTX NPs.Dialysis was used to evaluate the release rate of PTX in PTX NPs.Fluorescence inverted microscope was used to observe the uptake of PTX NPs.The CCK-8 method was used to detect the effect of NPs and PTX NPs on the growth of drug-resistant ovarian cancer cells.The animal imaging instrument was used to observe the biological distribution of PTX NPs in vivo.RESULTS The particle size of PTX NPs were(103.0±0.9)nm,and the encapsulation efficiency of PTX reached(84.1±1.3)%.There was a sequentially and controlled release of PTX within PTX NPs.The results of cellular uptake experiments showed that PTX NPs were more readily taken up by A2780 TR cells than free PTX.No-load mPEG-PLGA-PLL nanoparticles showed no obvious cytotoxicity.The IC50 of free PTX in A2780 and A2780 TR cells was(0.0102±0.0003)μg·mL^-1 and(4.55±0.44)μg·mL^-1,respectively.The IC50 of PTX NPs were(0.0031±0.0006)μg·mL^-1 and(1.05±0.13)μg·mL^-1,respectively.The results of in vivo biological distribution system showed that PTX NPs could target the tumor site.CONCLUSION The PTX NPs prepared in this study have suitable particle size,high encapsulation efficiency and sustained drug release.PTX NPs are easy to be taken up by cells.Compared with free PTX,PTX NPs have a stronger inhibitory effect on drug-resistant ovarian cancer cells.In vivo,PTX NPs have an effective tumor targeting effect.PTX NPs may have a promising application in the treatment of ovarian cancer.