【摘要】 目的:制备一种二硫键交联的精氨酸-天冬氨酸多肽聚合物(SRD),用作包载质粒PMEPA1(pPMEPA1,DPM)的载体,体外评价SRD/DPM复合物抗前列腺癌细胞迁移的疗效。方法:以低浓度的双氧水为氧化剂,半胱氨酸为交联剂,合成SRD,用1 H-NMR对其进行结构鉴定。将SRD与DPM按照不同氮磷比(N/P)涡旋混合,使其通过静电引力结合成SRD/DPM复合物。利用粒度电位分析仪测定SRD/DPM复合物的粒径和zeta电位,用琼脂糖凝胶电泳考察SRD对DPM的包载能力。将SRD和SRD/DPM复合物分别与前列腺癌细胞DU145共培养,考察细胞对复合物的摄取情况,以及复合物对癌细胞迁移能力的影响。结果:1H-NMR图谱显示SRD被成功合成。SRD与DPM通过静电引力结合,形成纳米复合物,表面呈正电性。凝胶电泳实验结果显示,N/P>2.5的SRD具有较强的基因药物包载能力。将SRD与DU145细胞共孵育24h后,细胞活力仍>50%。细胞摄取与Transwell实验显示,与RD单体相比,SRD能够显著增强DU145细胞对复合物的摄取,提高DPM抑制前列腺癌细胞迁移的效果。结论:以SRD作为基因药物载体传递DPM,可抑制前列腺癌细胞迁移,有望用于骨转移前列腺癌的治疗。更多还原

【Abstract】 Objective:To prepare a disulfide-bonded arginine-aspartic acid polypeptide polymer(SRD)as a gene delivery carrier of plasmid PMEPA1(pPMEPA1,DPM)and to evaluate its anti-prostate cancer cell migration ability in vitro.Methods:SRD was synthesized by using H2 O2 as oxidant and cysteine as cross linking agent.The structure of SRD was characterized by1 H-NMR.The SRD/DPM compounds were prepared by mixing SRD and DPM at various nitrogen/phosphorus(N/P)ratios.Zeta sizer was used to measure the particle size and zeta potential of SRD/DPM compounds,and agarose gel electrophoresis(AGE)was employed to determine the DPM condensation ability of SRD.Cytotoxicity of the carriers was measured through incubating SRD and SRD/DPM compounds with prostate cancer cell DU145.Cellular uptake efficiency of SRD/DPM compounds was measured by a flow cytometer using YOYO-1 labeled DPM as an indicator.Transwell assay was performed to determine cell migration ability of SRD/DPM compounds to DU145 cells in vitro.Results:1 H-NMR spectrum showed that the gene carrier SRD was successfully synthesized.SRD and DPM were combined to form a nanocomposite by electrostatic attraction,and the surface was positively charged.AGE results showed that SRD with N/P>2.5 had strong loading capacity to gene drugs.After incubating SRD with DU145 cells for 24 h,the cell viability was still more than 50%.Cell uptake and Transwell experiment indicated that SRD could significantly enhance the uptake of DPM by DU145 cells and improve the effects of DPM on the migration of DU145 cells when it was compared with RD monomer.Conclusion:Delivery of DPM with SRD as a gene drug carrier could inhibit prostate cancer cell migration and was expected to be a promising treatment of prostate cancer bone metastasis.更多还原