载阿霉素的TPGS-聚乙二醇-神经酰胺混合胶束对乳腺癌细胞多药耐药的逆转作用
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摘要
目的:构建载阿霉素的TPGS-聚乙二醇-神经酰胺混合胶束,逆转MCF-7/ADR细胞的多药耐药性。方法:采用薄膜分散法制备载阿霉素的TPGS-聚乙二醇-神经酰胺混合胶束,利用动态光散射法和透射电镜测定胶束的粒径与形态,并考察包封率、载药量和体外释放行为,利用CCK-8法测定游离阿霉素和载药胶束对MCF-7和MCF-7/ADR细胞的毒性作用。结果:载阿霉素的TPGS-聚乙二醇-神经酰胺混合胶束形态圆整,粒径为(15. 24±1. 72) nm,包封率为(96. 86±6. 12)%,载药量为(4. 72±0. 36)%,48 h的累积释放量为80%。游离阿霉素和载药胶束在48 h对MCF-7细胞的IC50值分别为0. 29和0. 30μg·m L-1,无统计学差异(P> 0. 05);对MCF-7/ADR细胞IC50值分别为47. 65和1. 61μg·m L-1,具有显著统计学差异(P <0. 001);载药胶束的逆转倍数为29. 61倍。结论:成功制备了粒径较小、分散均匀的载阿霉素的混合胶束,能有效逆转MCF-7/ADR细胞的多药耐药性。
Objective: To prepare doxorubicin( DOX)-loaded TPGS-polyethylene glycol-ceramide micelles and investigate the reversal of multidrug resistance in MCF-7/ADR cells. Methods: DOX-loaded TPGS-polyethylene glycol-ceramide micelles were prepared by lipid-film-based method. The size and morphology of the micelles were determined by dynamic light scattering and transmission electron microscopy. The encapsulation efficiency,drug loading and in vitro release were investigated. The cytotoxicity of free DOX and drug-loaded micelles against MCF-7 and MCF-7/ADR cells were determined by CCK-8. Results: The micelles were round in shape with a uniform size of( 15. 24 ± 1. 72) nm; the encapsulation efficiency and drug loading were( 96. 86 ± 6. 12) % and( 4. 72 ±0. 36) %,respectively; and the accumulative release was up to 80% at 48 h. The IC50 values of free DOX and drug-loaded micelles for MCF-7 cells at 48 h were 0. 29 and 0. 30 μg·m L~(-1),respectively,and the results between the two groups exhibited no significant difference( P > 0. 05),while the IC50 values for MCF-7/ADR cells at 48 hwere 47. 65 and 1. 61 μg·m L~(-1),respectively,and the results between the two groups exhibited significant difference( P < 0. 001). The reversal index was 29. 61. Conclusion: DOX-loaded polyethylene glycol-ceramide micelles with a small particle size and uniform dispersion were successfully prepared,which could effectively reverse the multidrug resistance of MCF-7/ADR cells.
Objective: To prepare doxorubicin( DOX)-loaded TPGS-polyethylene glycol-ceramide micelles and investigate the reversal of multidrug resistance in MCF-7/ADR cells. Methods: DOX-loaded TPGS-polyethylene glycol-ceramide micelles were prepared by lipid-film-based method. The size and morphology of the micelles were determined by dynamic light scattering and transmission electron microscopy. The encapsulation efficiency,drug loading and in vitro release were investigated. The cytotoxicity of free DOX and drug-loaded micelles against MCF-7 and MCF-7/ADR cells were determined by CCK-8. Results: The micelles were round in shape with a uniform size of( 15. 24 ± 1. 72) nm; the encapsulation efficiency and drug loading were( 96. 86 ± 6. 12) % and( 4. 72 ±0. 36) %,respectively; and the accumulative release was up to 80% at 48 h. The IC50 values of free DOX and drug-loaded micelles for MCF-7 cells at 48 h were 0. 29 and 0. 30 μg·m L~(-1),respectively,and the results between the two groups exhibited no significant difference( P > 0. 05),while the IC50 values for MCF-7/ADR cells at 48 hwere 47. 65 and 1. 61 μg·m L~(-1),respectively,and the results between the two groups exhibited significant difference( P < 0. 001). The reversal index was 29. 61. Conclusion: DOX-loaded polyethylene glycol-ceramide micelles with a small particle size and uniform dispersion were successfully prepared,which could effectively reverse the multidrug resistance of MCF-7/ADR cells.
引文
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[13] BAO Y,YIN M,HU X,et al. A safe,simple and efficient doxorubicin prodrug hybrid micelle for overcoming tumor multidrug resistance and targeting delivery[J]. J Control Release,2016,235:182-194.
[14] YANG C,WU T,QI Y,et al. Recent advances in the application of vitamin E TPGS for drug delivery[J]. Theranostics,2018,8(2):464-485.
[15] SU X,SONG H,NIU F,et al. Co-delivery of doxorubicin and PEGylated C16-ceramide by nanoliposomes for enhanced therapy against multidrug resistance[J]. Nanomedicine(Lond),2015,10(13):2033-2050.
[16]王美平,钟延强,高洁,等.载阿霉素的聚乙二醇-神经酰胺胶束的制备和表征[J].中国新药杂志,2016,25(6):716-720.
[17] WANG M,XIE F,WEN X,et al. Therapeutic PEG-ceramide nanomicelles synergize with salinomycin to target both liver cancer cells and cancer stem cells[J]. Nanomedicine(Lond),2017,12(9):1025-1042.
[2]仇笳熙,卿晨.肿瘤多药耐药机制的研究进展[J].实用肿瘤学杂志,2015,29(4):364-367.
[3] KARTAL-YANDIM M,ADAN-GOKBULUT A,BARAN Y. Molecular mechanisms of drug resistance and its reversal in cancer[J]. Crit Rev Biotechnol,2016,36(4):716-726.
[4] RATH G,SCHNEIDER C,LANGLOIS B,et al. De novo ceramide synthesis is responsible for the anti-tumor properties of camptothecin and doxorubicin in follicular thyroid carcinoma[J].Int J Biochem Cell Bio,2009,41(5):1165-1172.
[5] DUMITRU CA,CARPINTEIRO A,TRARBACH T,et al.Doxorubicin enhances TRAIL-induced cell death via ceramideenriched membrane platforms[J]. Apoptosis,2007,12(8):1533-1541.
[6] LIU YY,HILL RA,LI YT. Ceramide glycosylation catalyzed by glucosylceramide synthase and cancer drug resistance[J]. Adv Cancer Res,2013,117:59-89.
[7]戴东波,徐金中,尤文挺,等.盐酸阿霉素聚乳酸纳米粒的制备及大鼠体内药动学研究[J].中国现代应用药学,2017,34(8):1127-1132.
[8] PATEL NR,PATTNI BS,ABOUZEID AH,et al. Nanopreparations to overcome multidrug resistance in cancer[J]. Adv Drug Deliv Rev,2013,65(13-14):1748-1762.
[9] KAPSE-MISTRY S,GOVENDER T,SRIVASTAVA R,et al.Nanodrug delivery in reversing multidrug resistance in cancer cells[J]. Front Pharmacol,2014,5:159.
[10] SHAPIRA A,LIVNEY YD,BROXTERMAN HJ,et al. Nanomedicine for targeted cancer therapy:towards the overcoming of drug resistance[J]. Drug Resist Updat,2011,14(3):150-163.
[11] GUPTA P,GARG T,TANMAY M,et al. Polymeric drug-delivery systems:role in P-gp efflux system inhibition[J]. Crit Rev Ther Drug Carrier Syst,2015,32(3):247-275.
[12] ZHU H,CHEN H,ZENG X,et al. Co-delivery of chemotherapeutic drugs with vitamin E TPGS by porous PLGA nanoparticles for enhanced chemotherapy against multi-drug resistance[J].Biomaterials,2014,35(7):2391-2400.
[13] BAO Y,YIN M,HU X,et al. A safe,simple and efficient doxorubicin prodrug hybrid micelle for overcoming tumor multidrug resistance and targeting delivery[J]. J Control Release,2016,235:182-194.
[14] YANG C,WU T,QI Y,et al. Recent advances in the application of vitamin E TPGS for drug delivery[J]. Theranostics,2018,8(2):464-485.
[15] SU X,SONG H,NIU F,et al. Co-delivery of doxorubicin and PEGylated C16-ceramide by nanoliposomes for enhanced therapy against multidrug resistance[J]. Nanomedicine(Lond),2015,10(13):2033-2050.
[16]王美平,钟延强,高洁,等.载阿霉素的聚乙二醇-神经酰胺胶束的制备和表征[J].中国新药杂志,2016,25(6):716-720.
[17] WANG M,XIE F,WEN X,et al. Therapeutic PEG-ceramide nanomicelles synergize with salinomycin to target both liver cancer cells and cancer stem cells[J]. Nanomedicine(Lond),2017,12(9):1025-1042.