Abstract：
Pitavastatin (PITA) is a 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase inhibitor to treat hypercholesterolemia and in recent studies is focused that its potential anti-cancer effect. This study was aimed to elucidate the effect of PITA alone and in combination with cisplatin on cervical cancer cells (HeLa) in vitro. Cytotoxicity of PITA (5-200 μM) was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and neutral red uptake (NRU) assays for 24, 48, and 72 h. Cell apoptosis and cell cycle analyses were performed in flow cytometry (0.1-100 μM). The evaluation of genotoxic effects and oxidative DNA damage of PITA (2-200 μM) were performed with standard comet assay, formamidopyrimidine glycosylase (fpg)-modified comet assay, and reactive oxygen species (ROS) activation in HeLa cells. PITA alone reduced cell viability in a dose-dependent manner (20-200, 20-200, and 5-200 μM for 24, 48, and 72 h, respectively, in MTT). The combined treatment of PITA with cisplatin resulted in significantly greater inhibition of cell viability. ROS and DNA damage increased significantly at 100 μM for 4 h and 20 μM for 24 h, respectively. PITA-induced apoptosis, an increased proportion of sub G1 cells, was monitored, and also, it increased the expression of active caspase-9 and caspase-3 and upregulated cleaved poly adenosine diphosphate ribose polymerase (PARP) by western blotting and caspase 3/8/9 multiple assay kit. We conclude that PITA can be used to efficiently cervical cancer studies, and promising findings have been obtained for further studies.
摘要：
匹伐他汀（PITA）是一种3-羟基-3-甲基戊二酰辅酶A（HMG-CoA）还原酶抑制剂，用于治疗高胆固醇血症，最近的研究集中于其潜在的抗癌作用。本研究旨在阐明PITA单独和联合顺铂对宫颈癌细胞（HeLa）的体外作用。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物(MTT)和中性红摄取(NRU)测定24、48和72小时来评估PITA(5-200μM)的细胞毒性。在流式细胞术(0.1-100μM)中进行细胞凋亡和细胞周期分析。使用标准彗星测定法对PITA（2-200μM）的遗传毒性作用和氧化DNA损伤进行评估，甲酰氨基嘧啶糖基化酶（fpg）修饰的彗星试验，和HeLa细胞中的活性氧（ROS）活化。PITA单独以剂量依赖性方式降低细胞活力（20-200、20-200和5-200μM持续24、48和72小时，分别,在MTT中）。PITA与顺铂的联合治疗导致显著更大的细胞活力抑制。ROS和DNA损伤在100μM持续4小时和20μM持续24小时时显著增加，分别。PITA诱导的细胞凋亡,亚G1细胞比例增加，被监控,而且,通过蛋白质印迹和caspase3/8/9多重分析试剂盒，它增加了活性caspase-9和caspase-3的表达，并上调了裂解的聚二磷酸腺苷核糖聚合酶（PARP）。我们得出结论，PITA可以用于有效的宫颈癌研究，并获得了有希望的发现供进一步研究。
