四丁基丙二胺对人MG63骨髓瘤细胞增殖、凋亡和迁移能力的影响

Effects of polyamine analogues tetrabutyl propanediamine on proliferation,apoptosis and migration of human MG63 myeloma cells

目的分析新多胺类似物四丁基丙二胺(tetrabutylpropanediamine,TBP)对人骨髓瘤MG63细胞生长的影响及其机制。方法 MTT法用于分析细胞增殖,流式细胞分析用于细胞周期和凋亡细胞鉴定,琼脂糖凝胶电泳用于分析DNA片段化,Western blot用于分析细胞凋亡相关蛋白的表达水平,Transwell技术用于分析细胞的迁移能力。结果TBP明显抑制MG63细胞的增殖和迁移能力,抑制效应呈时间和剂量依赖性。流式细胞分析发现,TBP干扰细胞周期,导致G1和G2期细胞比例升高,但S期细胞比例明显下降,同时凋亡细胞数量大幅增加。TBP处理后,细胞染色体DNA出现凋亡细胞典型的DNA片段化现象,细胞质中促凋亡蛋白Bax和细胞色素C含量明显升高。结论 TBP具有抑制人骨髓瘤MG63细胞增殖的药理活性,其机制与抑制细胞周期和诱导细胞凋亡有关,提示TBP具有用于临床骨髓瘤治疗的潜在价值。

Aim To study the effects of the new polyamine analogue tetrabutyl propanediamine（TBP） on proliferation,apoptosis and migration of human myeloma MG63 cells and its mechanism.Methods MTT was used for the analysis of cell proliferation.The flow cytometry was performed to identify cell cycle and apoptotic cells.The agarose-gel electrophoresis was used to evaluate DNA fragmentation.Western blot was used to identify the expression of apoptosis-related protein.Transwell technique was used for the analysis of cell migration.Results TBP inhibited proliferation and migration of MG63 cell significantly and the inhibitory effect was in a time-and dose-dependent manner.The results from flow cytometry analysis indicated that TBP interfered with cell cycles,leading to increased cell percentages in G1 and G2 phase,but decreased cell percentage in S phase.In the same time,the number of apoptotic cells increased significantly.After TBP treatment,DNA fragmentation of chromosomal DNA,typical for apoptotic cells,was observed and the contents of pro-apoptotic protein Bax and cytochrome C in the cytosol were significantly increased.Conclusion TBP can inhibit the proliferation of human myeloma MG63 cells by inhibiting cell cycle and inducing apoptosis,suggesting a potential value of TBP in clinical treatment of myeloma.