2-氯脱氧腺苷(2-CDA)对人黑色素瘤细胞系A375生物学性质的影响

doi:10.3971/j.issn.1000-8578.2011.09.004

2-氯脱氧腺苷(2-CDA)对人黑色素瘤细胞系A375生物学性质的影响

Effect of 2-chloro-2′-deoxyadenosine (2-CDA) on Biological Property in Human Melanoma A375 Cells

摘要

摘要: 目的观察2-CDA对人黑色素瘤细胞系A375细胞在增殖、形态、周期、迁移和凋亡等生物学性质的影响。方法MTT 比色法检测2-CDA对A375细胞增殖的影响；瑞氏-姬姆萨染色观察细胞形态的变化；流式细胞仪分析细胞周期及细胞凋亡的改变；划痕修复实验检测A375细胞迁移能力的变化；蛋白印迹检测细胞内凋亡相关蛋白的变化情况。结果2-CDA 呈时间-剂量依赖性地抑制A375细胞的增殖；瑞氏-姬姆萨染色显示2-CDA处理后，细胞形态不规则。流式细胞仪检测表明2-CDA能够诱导A375细胞阻滞于S期。划痕修复实验结果显示2-CDA能够显著抑制A375细胞的迁移能力；Annexin-V/PI双标法检测结果表明2-CDA可诱导细胞凋亡；Western blot检测结果显示，2-CDA作用A375细胞后，Caspase-3被剪切活化，其底物蛋白PARP发生剪切。结论2-CDA能够对A375细胞增殖、形态、周期、迁移等生物学性质产生广泛的影响，并激活Caspase-3信号通路诱导细胞凋亡。

Abstract: ObjectiveTo investigate the effect of 2-chloro-2'-deoxyadenosine (2-CDA) on the proliferation,morphology,cell cycle,migration and apoptosis in human melanoma A375 cells. Methods The inhibitory effect of 2-CDA on A375 proliferation was detected by MTT assay.Morphological change was observed by Giemsa staining.The cell cycle arrest and apoptosis was determined by flow cytometry.Migration rate was analyzed by scratch wound healing assays.Protein levels were determined by western blot. Results 2-CDA inhibited the proliferation of A375 cells in both time- and dose-dependent manner.Giemsa staining showed that after treatment of 2-CDA,the cellular morphology became irregular.A375 cells were induced to arrest at S phases.Scratch wound healing assay showed that 2-CDA obviously inhibited the migration of A375 cells. Results of Western blot showed that after treatment of 2-CDA for 36 h,Caspase-3 and PARP were all cleaved. Conclusion 2-CDA can effectively influence the proliferation,morphology,cell cycle,migration in human melanoma A375 cells,and induce apoptosis through caspase3 pathway.

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