Effects of the New Polyamine Analog Tetrabutyl Propanediamine on the Apoptosis and Migration of Human Bladder Carcinoma T24 Cells
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摘要:
目的 研究新多胺类似物四丁基丙二胺(tetrabutyl propaneadiamine, TBP)对人膀胱移行细胞癌T24细胞生长、凋亡及迁移能力的影响。 方法 MTT比色法分析细胞增殖, 流式细胞术分析细胞周期改变, TUNEL法检测细胞凋亡, Western blot用于分析细胞凋亡相关蛋白Bax和细胞色素C的表达水平, Transwell技术检测细胞迁移能力的变化。 结果 TBP有效抑制T24细胞增殖及迁移能力。细胞周期分析显示TBP抑制细胞发生G_1/S期转换, 导致S期细胞比例显著性下降, G_0/G_1期细胞比例升高, 同时出现SubG_1亚凋亡峰。TBP处理后, 细胞呈现典型凋亡现象, 细胞浆中促凋亡蛋白Bax和细胞色素C含量明显升高。 结论 TBP能够抑制人膀胱移行细胞癌T24细胞增殖、降低T24细胞迁移能力, 诱导细胞凋亡, 其机制可能与影响肿瘤细胞的细胞周期, 活化线粒体相关的细胞凋亡途径有关。 Abstract:Objective This work aimed to investigate the effects of the new polyamine analog tetra-butyl propanediamine (TBP) on the proliferation, apoptosis, and migration of human bladder carcinoma T24 cells and its potential mechanism. Methods Methyl thiazolyl tetrazolium was used to analyze cell proliferation, and flow cytometry was performed to assay the cell cycles. Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling assay was used to identify the apoptotic cells. Western blot analysis was used to evaluate the expression of apoptosis-related proteins. The Transwell technique was used to analyze cell migration. Results TBP significantly inhibited the proliferation and migration of T24 cells, and the inhibitory effect was time and dose dependent. The flow cytometry results indicated that TBP interfered with the cell cycles and inhibited G1/S transition, which led to increased cell percentage in the G1 and G2 phases, as well as decreased cell percentage in the S phase. Meanwhile, the number of apoptotic cells significantly increased. After TBP treatment, observation of the typical behavior of apoptotic cells revealed that the contents of the pro-apoptotic proteins Bax and cytochrome C significantly increased in the cytosol. Conclusion TBP can inhibit the proliferation and induce the apoptosis of human bladder carcinoma T24 cells by interfering with the cell cycle and activating the mitochondrion-mediated apoptotic pathway. -
Key words:
- Polyamine analog /
- Tetra-butyl propanediamine /
- Bladder carcinoma /
- Migration /
- Apoptosis
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图 1 TBP抑制T24细胞存活率的比较（*P < 0.05，**P < 0.01）
Figure 1. Inhibitory action of TBP on the growth of T24 cells, compared with the 0 mol/L group
图 2 Transwell技术检测TBP对T24细胞迁移能力的影响（×200）
A1，A2：未处理的T24细胞；B1，B2：40 μmol/L TBP处理的T24细胞
Figure 2. Results of the Transwell chamber assay for T24 cells
图 3 TUNEL法检测T24细胞凋亡
Figure 3. TUNEL assay was performed to observe apoptosis of T24 cells
图 4 Western blot检测T24细胞的胞浆内Bax和细胞色素C水平
1：未处理的T24细胞；2：40 umol/L TBP处理48 h的T24细胞
Figure 4. Western blot analysis results of the expression of Bax and Cyt-C proteins in the cytosol of T24 cells
表 1 TBP对T24细胞周期的影响
Table 1. Effects of TBP on the cell cycle and apoptosis of T24 cells
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