Movafegh Bahareh, Jalal Razieh, Mohammadi Zobeideh, Aldaghi Seyyede A
Department of Chemistry, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.
Cell and Molecular Biotechnology Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Anticancer Agents Med Chem. 2018;18(10):1448-1456. doi: 10.2174/1871520618666180412114750.
Cell resistance to doxorubicin and its toxicity to healthy tissue reduce its efficiency. The use of cell-penetrating peptides as drug delivery system along with doxorubicin is a strategy to reduce its side effects. In this study, the influence of poly-L-arginine on doxorubicin cytotoxicity, its cellular uptake and doxorubicin-induced apoptosis on human prostate cancer DU145 cells are assessed.
The cytotoxicity of doxorubicin and poly-L-arginine, alone and in combination, in DU145 cells was evaluated at different exposure times using MTT assay. The influence of poly-L-arginine on doxorubicin delivery into cells was evaluated by fluorescence microscopy and ultraviolet spectroscopy. DAPI and ethidium bromide- acridine orange stainings, flow cytometry using annexin V/propidium iodide, western blot analysis with anti-p21 antibody and caspase-3 activity were used to examine the influence of poly-L-arginine on doxorubicininduced cell death.
Poly-L-arginine had no cytotoxicity at low concentrations and short exposure times. Poly-L-arginine increased the cytotoxic effect of doxorubicin in DU145 cells in a time-dependent manner. But no significant reduction was found in HFF cell viability. Poly-L-arginine seems to facilitate doxorubicin uptake and increase its intracellular concentration. 24h combined treatment of cells with doxorubicin (0.5 µM) and poly-L-arginine (1 µg ml-1) caused a small increase in doxorubicin-induced apoptosis and significantly elevated necrosis in DU145 cells as compared to each agent alone.
Our results indicate that poly-L-arginine at lowest and highest concentrations act as proliferationinducing and antiproliferative agents, respectively. Between these concentrations, poly-L-arginine increases the cellular uptake of doxorubicin and its cytotoxicity through induction of necrosis.
细胞对阿霉素的耐药性及其对健康组织的毒性降低了其疗效。使用细胞穿透肽作为药物递送系统与阿霉素联合使用是一种降低其副作用的策略。在本研究中，评估了聚-L-精氨酸对阿霉素细胞毒性、其细胞摄取以及阿霉素诱导人前列腺癌DU145细胞凋亡的影响。
使用MTT法在不同暴露时间评估阿霉素和聚-L-精氨酸单独及联合作用于DU145细胞的细胞毒性。通过荧光显微镜和紫外光谱评估聚-L-精氨酸对阿霉素进入细胞的影响。使用DAPI和溴化乙锭-吖啶橙染色、膜联蛋白V/碘化丙啶流式细胞术、抗p21抗体的蛋白质免疫印迹分析和半胱天冬酶-3活性来检测聚-L-精氨酸对阿霉素诱导的细胞死亡的影响。
聚-L-精氨酸在低浓度和短暴露时间下无细胞毒性。聚-L-精氨酸以时间依赖性方式增加了阿霉素对DU145细胞的细胞毒性作用。但在人包皮成纤维细胞（HFF）活力方面未发现显著降低。聚-L-精氨酸似乎促进了阿霉素的摄取并增加了其细胞内浓度。与单独使用每种药物相比，用阿霉素（0.5 μM）和聚-L-精氨酸（1 μg/ml）联合处理细胞24小时导致阿霉素诱导的DU145细胞凋亡略有增加，坏死显著升高。
我们的结果表明，最低和最高浓度的聚-L-精氨酸分别作为增殖诱导剂和抗增殖剂。在这些浓度之间，聚-L-精氨酸通过诱导坏死增加阿霉素的细胞摄取及其细胞毒性。
