Kaplan Ayse, Akalin Ciftci Gulsen, Kutlu Hatice Mehtap
Department of Biology, Institute of Science, Anadolu University, 26470, Eskisehir, Turkey.
Department of Biochemistry, Faculty of Pharmacy, Anadolu University, 26470, Eskisehir, Turkey.
Cytotechnology. 2016 Oct;68(5):1727-35. doi: 10.1007/s10616-015-9922-5. Epub 2015 Oct 26.
Metal based drugs have successfully been used in both the detection and treatment of different disease states. The antibacterial features of metal ion silver are well documented. Most recently, metal ion silver has been tested and applied in anticancer activity. The present study observed the cytotoxic, anti-proliferative and apoptotic effects of metal complex silver nitrate in H-ras transformed 5RP7 cell lines for 24 h. In addition, the toxic effects of silver nitrate was investigated on NIH/3T3 primary mouse embryonic fibroblast cells for 24 h. Cytotoxic effects were determined by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay. Apoptosis and necrosis were evaluated by flow cytometric analysis (Annexin-V FITC/PI). Caspase-3 activation was researched by flow cytometric analysis. Apoptotic morphology was observed by DAPI staining. Structure and ultra-structure changes of cells were assessed using transmission electron microscopy. The results indicate silver nitrate has high cytotoxicity and a strong capacity to induce apoptosis in H-ras 5RP7 cells. Furthermore silver nitrate was not toxic against NIH/3T3 primary mouse embryonic fibroblast cells at low doses for 24 h.
金属基药物已成功用于不同疾病状态的检测和治疗。金属离子银的抗菌特性已有充分记录。最近，金属离子银已在抗癌活性方面进行了测试和应用。本研究观察了金属络合物硝酸银在H-ras转化的5RP7细胞系中24小时的细胞毒性、抗增殖和凋亡作用。此外，还研究了硝酸银对NIH/3T3原代小鼠胚胎成纤维细胞24小时的毒性作用。细胞毒性作用通过MTT（3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四唑溴盐）法测定。通过流式细胞术分析（膜联蛋白-V FITC/PI）评估凋亡和坏死情况。通过流式细胞术分析研究半胱天冬酶-3的激活情况。通过DAPI染色观察凋亡形态。使用透射电子显微镜评估细胞的结构和超微结构变化。结果表明，硝酸银在H-ras 5RP7细胞中具有高细胞毒性和强大的诱导凋亡能力。此外，低剂量的硝酸银在24小时内对NIH/3T3原代小鼠胚胎成纤维细胞无毒。
