Mazur Antonina Joanna, Nowak Dorota, Mannherz Hans Georg, Malicka-Błaszkiewicz Maria
Department of Anatomy and Embryology, Faculty of Medicine, Ruhr University, Bochum, Germany.
Eur J Pharmacol. 2009 Jun 24;613(1-3):24-33. doi: 10.1016/j.ejphar.2009.04.020. Epub 2009 Apr 19.
Methotrexate is a widely used drug in treatments of various types of malignancies and in the therapy of rheumatoid arthritis. The goal of our study was to look at the effect of this dihydrofolate reductase inhibitor on the actin cytoskeleton, since actin plays an important role in cancer transformation and metastasis. For this reason we compared results obtained from experiments on CaSki (human uterine cervix cancer) and NRK (normal fibroblastic rat kidney) cells treated with methotrexate. It has been shown previously that methotrexate can induce apoptosis. Therefore we first examined whether methotrexate induces apoptosis in our model cells. For this aim we applied several assays like Caspase Glo 3/7, DNA fragmentation and binding of phosphatidylserine by annexin V-fluorescein. The data obtained indicated that methotrexate induces programmed cell death in CaSki and NRK cells. However, differences between CaSki and NRK cells were observed in the morphological alterations and dynamics of apoptosis induced by methotrexate. It seemed that cancer cells were more sensitive towards the cell death inducing activity at lower concentrations of methotrexate. Analysis by confocal microscopy of methotrexate-treated cells demonstrated that treatment with this folate antagonist affected the actin cytoskeleton, although the dis-organization of the actin cytoskeleton after treatment with methotrexate differed between cancer and normal cells.
甲氨蝶呤是一种广泛应用于各类恶性肿瘤治疗及类风湿关节炎治疗的药物。我们研究的目的是探究这种二氢叶酸还原酶抑制剂对肌动蛋白细胞骨架的影响，因为肌动蛋白在癌症转化和转移过程中起着重要作用。因此，我们比较了用甲氨蝶呤处理的CaSki（人子宫颈癌）细胞和NRK（正常大鼠肾成纤维细胞）细胞的实验结果。先前已经表明甲氨蝶呤可诱导细胞凋亡。所以我们首先检测甲氨蝶呤在我们的模型细胞中是否诱导细胞凋亡。为此目的，我们应用了几种检测方法，如Caspase Glo 3/7检测、DNA片段化检测以及膜联蛋白V-荧光素结合磷脂酰丝氨酸检测。所获得的数据表明甲氨蝶呤在CaSki和NRK细胞中诱导程序性细胞死亡。然而，在甲氨蝶呤诱导的凋亡的形态学改变和动力学方面，观察到CaSki细胞和NRK细胞之间存在差异。似乎在较低浓度的甲氨蝶呤作用下，癌细胞对诱导细胞死亡的活性更敏感。对经甲氨蝶呤处理的细胞进行共聚焦显微镜分析表明，用这种叶酸拮抗剂处理会影响肌动蛋白细胞骨架，尽管甲氨蝶呤处理后肌动蛋白细胞骨架的紊乱在癌细胞和正常细胞之间有所不同。
