负载紫杉醇CSNRDARRC-PCL-PGA/TPGS多肽纳米颗粒调控膀胱癌RT112细胞增生及促凋亡
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摘要
目的探讨双靶向多肽CSNRDARRC-PCL-PGA/TPGS同时负载紫杉醇的纳米颗粒(多肽紫杉醇NP)对膀胱癌RT112细胞的生长抑制及促凋亡作用。方法通过CSNRDARRCPCL-PGA与聚乙二醇1000维生素E琥珀酸酯(TPGS)形成共混胶束再负载紫杉醇(多肽紫杉醇NP),其后通过MTT检测细胞存活、DAPI及Annex V/PI染色分析细胞凋亡、JC-1腺粒体膜电位分析检测多肽紫杉醇NP对膀胱癌RT112细胞的影响。结果多肽紫杉醇NP能够抑制RT112细胞生长,并且有时间依赖性;细胞周期分析显示RT112细胞停滞在G2期,DAPI及Annex V/PI染色均可见细胞凋亡;JC-1染色发现多肽紫杉醇NP是通过腺粒体膜电位下降引起细胞凋亡。所有结果均显示多肽紫杉醇NP优于紫杉醇NP。结论 CSNRDARRC多肽修饰的TPGS-b-(PCLran-PGA)负载紫杉醇纳米颗粒(多肽紫杉醇NP)抑制膀胱癌RT112细胞增生及促进凋亡,为临床治疗膀胱癌提供了一个新手段。
Objective To investigate the effect of Taxol loaded-nanoparticles of double targeting CSNRDARRC-PCL-PGA/TPGS(Polypeptide Taxol NP)on cell proliferation inhibition and apoptosis of bladder carcinoma cell line RT112. Methods Taxol loaded-nanoparticles were prepared by self-assembly of CSNRDARRC-PCL-PGA-α-tocopheryl polyethylene glycol 1000 succinate[TP〗.Then the cell viability was assessed using an MTT assay,the cell apoptosis was assessed using DAPI staining and Annex V/PI.To analyze that Polypeptide Taxol NP affected the mitochondria membrane potential by JC-1 staining in bladder carcinoma cell line RT112. Results Polypeptide Taxol NP could inhibited cell proliferation in time-dependent manner.In cell cycle assay,Polypeptide Taxol NP promoted G2 phase arrest.Then the occurrence of apoptosis in Annex V/PI staining and DAPI staining.Finally,Polypeptide Taxol NP induced apoptosis through decreasing mitochondria membrane potential in JC-1 staining.All results show that Polypeptide Taxol NP is better than Polypeptide Taxol NP,because Polypeptide is targeted at bladder carcinoma. Conclusions Taxol loadednanoparticles of CSNRDARRC-PCL-PGA/TPGS(Polypeptide Taxol NP)exhibits anti-tumor effect on RT112 cells by inhibiting proliferation and promoting apoptosis.And it provides a novel powerful nanomedicine platform for bladder carcinoma.
Objective To investigate the effect of Taxol loaded-nanoparticles of double targeting CSNRDARRC-PCL-PGA/TPGS(Polypeptide Taxol NP)on cell proliferation inhibition and apoptosis of bladder carcinoma cell line RT112. Methods Taxol loaded-nanoparticles were prepared by self-assembly of CSNRDARRC-PCL-PGA-α-tocopheryl polyethylene glycol 1000 succinate[TP〗.Then the cell viability was assessed using an MTT assay,the cell apoptosis was assessed using DAPI staining and Annex V/PI.To analyze that Polypeptide Taxol NP affected the mitochondria membrane potential by JC-1 staining in bladder carcinoma cell line RT112. Results Polypeptide Taxol NP could inhibited cell proliferation in time-dependent manner.In cell cycle assay,Polypeptide Taxol NP promoted G2 phase arrest.Then the occurrence of apoptosis in Annex V/PI staining and DAPI staining.Finally,Polypeptide Taxol NP induced apoptosis through decreasing mitochondria membrane potential in JC-1 staining.All results show that Polypeptide Taxol NP is better than Polypeptide Taxol NP,because Polypeptide is targeted at bladder carcinoma. Conclusions Taxol loadednanoparticles of CSNRDARRC-PCL-PGA/TPGS(Polypeptide Taxol NP)exhibits anti-tumor effect on RT112 cells by inhibiting proliferation and promoting apoptosis.And it provides a novel powerful nanomedicine platform for bladder carcinoma.
引文
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[2]李路,杨军,李兰,等.罗勒多糖联合紫杉醇对A549细胞线粒体膜电位和细胞周期的影响[J].中南药学,2016,(5):471-475.
[3]Xiao M,Hong Z,Sun L,et al.TMTP1,a Novel Tumorhoming Peptide,Specifically Targets Hematological Malignancies and Their Metastases[J].J Huazhong U Sci-Med,2011,31(5):608-613.
[4]Jia XY,Yu Q,Zhang ZH,et al.Targeting bladder tumor cells in voided urine of Chinese patients with FITC-CSNRDARRC peptide ligand[J].Onco Targets Ther,2012,5:85-90.
[5]Yao VJ,D'Angelo S,Butler KS,et al.Ligand-targeted theranostic nanomedicines against cancer[J].J Control Release,2016,240:267-286.
[6]Yang XF,Pang JZ,Liu JH,et al.Homing Peptide Guiding Optical Molecular Imaging for the Diagnosis of Bladder Cancer[J].Proc Spie,2014,(96281):1-12.
[7]Dintaman JM,Silverman JA.Inhibition of P-glycoprotein by D-alpha-tocopheryl polyethylene glycol 1000 succinate(TPGS)[J].Pharm Res,1999,16(10):1550-1556.
[8]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.
[9]Sun Y,Yu B,Wang G,et al.Enhanced antitumor efficacy of vitamin E TPGS-emulsified PLGA nanoparticles for delivery of paclitaxel[J].Colloids Surf B Biointerfaces,2014,123:716-723.
[10]Mu L,Feng SS.A novel controlled release formulation for the anticancer drug paclitaxel(Taxol):PLGA nanoparticles containing vitamin E TPGS[J].J Control Release,2003,86(1):33-48.
[11]Watanabe T,Miyauchi S,Sawada Y,et al.Kinetic-Analysis of Hepatobiliary Transport of Vincristine in Perfused-RatLiver-Possible Roles of P-Glycoprotein in Biliary-Excretion of Vincristine[J].J Hepatol,1992,16(1-2):77-88.
[12]Choudhury H,Gorain B,Pandey M,et al.Recent advances in TPGS-based nanoparticles of docetaxel for improved chemotherapy[J].Int J Pharmaceut,2017,529(1-2):506-522.
[13]Duhem N,Rolland J,Riva R,et al.Tocol modified glycol chitosan for the oral delivery of poorly soluble drugs[J].Int J Pharm,2012,423(2):452-460.
[14]Ma Y,Zheng Y,Zeng X,et al.Novel docetaxel-loaded nanoparticles based on PCL-Tween 80 copolymer for cancer treatment[J].Int J Nanomedicine,2011,6:2679-2688.
[15]Huerta S,Goulet EJ,Huerta-Yepez S,et al.Screening and detection of apoptosis[J].J Surg Res,2007,139(1):143-156.
[16]Elmore S.Apoptosis:A review of programmed cell death[J].Toxicol Pathol,2007,35(4):495-516.
[17]Ouyang L,Shi Z.,Zhao S,et al.Programmed cell death pathways in cancer:a review of apoptosis,autophagy and programmed necrosis[J].Cell Prolif,2012,45(6):487-498.
[18]Jiang X,Wang X.Cytochrome C-mediated apoptosis[J].Annu Rev Biochem,2004,73:87-106.
[19]Kroemer G,Reed JC.Mitochondrial control of cell death[J].Nat Med,2000,6(5):513-519.