在本研究中,我们研究了含Myc诱饵寡核苷酸的靶向甲氨蝶呤-硒纳米结构与X线照射联合治疗对LNCaP前列腺癌细胞的协同作用.基于Bcl-2基因启动子设计Myc诱饵ODN,并通过分子对接和分子动力学测定进行分析。将ODN加载到合成的Se@BSA@Chi-MTX纳米结构上。通过FTIR测定了纳米结构的物理化学特性,DLS,UV-vis,TEM,EDX,体外释放,和溶血试验。随后,通过吸收研究了它们在有和没有X射线照射下的细胞毒性特性,MTT,细胞周期,凋亡,和对LNCaP细胞系的划痕测定。DLS和TEM结果显示Se@BSA@Chi-DEC-MTXNPs的负电荷(-9mV)和纳米尺寸(40nm),分别。FTIR的结果,UV-vis,和EDX显示了不同部分的适当相互作用和纳米颗粒的正确合成。溶血结果显示浓度小于6mg/mL的该纳米颗粒的血液相容性。从纳米结构释放的ODN显示出pH依赖性的方式,在酸性pH下释放速率高15%。靶向的Se@BSA@Chi标记的ODN-MTXNP通过靶向前列腺特异性膜抗原(PSMA)被LNCaP细胞有效吸收。纳米结构(含MTX药物)处理与X-照射的显著协同作用显示细胞生长抑制,凋亡诱导(~57%),细胞周期停滞(G2/M期),与对照相比,迁移抑制(高达90%)。结果表明,Se@BSA@Chi-DEC-MTXNPs可以潜在地抑制LNCaP细胞的生长。这种纳米结构系统可以成为前列腺癌治疗中靶向药物递送和放化疗的有前途的方法。
In the present study, we investigated the synergistic effects of targeted methotrexate-selenium nanostructure containing Myc decoy oligodeoxynucleotides along with X-irradiation exposure as a combination therapy on LNCaP prostate cancer cells. Myc decoy ODNs were designed based on the promoter of Bcl-2 gene and analyzed by molecular docking and molecular dynamics assays. ODNs were loaded on the synthesized Se@BSA@Chi-MTX nanostructure. The physicochemical characteristics of nanostructures were determined by FTIR, DLS, UV-vis, TEM, EDX, in vitro release, and hemolysis tests. Subsequently, the cytotoxicity properties of them with and without X-irradiation were investigated by uptake, MTT, cell cycle, apoptosis, and scratch assays on the LNCaP cell line. The results of DLS and TEM showed negative charge (-9 mV) and nanometer size (40 nm) for Se@BSA@Chi-DEC-MTX NPs, respectively. The results of FTIR, UV-vis, and EDX showed the proper interaction of different parts and the correct synthesis of nanoparticles. The results of hemolysis showed the hemocompatibility of this nanoparticle in concentrations less than 6 mg/mL. The ODNs release from the nanostructures showed a pH-dependent manner, and the release rate was 15% higher in acidic pH. The targeted Se@BSA@Chi-labeled ODN-MTX NPs were efficiently taken up by LNCaP cells by targeting the prostate-specific membrane antigen (PSMA). The significant synergistic effects of nanostructure (containing MTX drug) treatment along with X-irradiation showed cell growth inhibition, apoptosis induction (~57%), cell cycle arrest (G2/M phase), and migration inhibition (up to 90%) compared to the control. The results suggested that the Se@BSA@Chi-DEC-MTX NPs can potentially suppress the cell growth of LNCaP cells. This nanostructure system can be a promising approach for targeted drug delivery and chemoradiotherapy in prostate cancer treatment.