替尼泊苷诱导Tca8113细胞凋亡及细胞周期阻滞的实验观察

Teniposide inducing apoptosis and G2/M phase arrest of Tca8113 cells

目的:检测替尼泊苷(teniposide,VM-26)的体外抗肿瘤增殖效果并探索其作用机制。方法:以不同浓度的VM-26体外作用Tca8113细胞,应用MTT方法、流式细胞仪、透射电镜和荧光染色等分别检测细胞的生长抑制率、凋亡率及细胞周期分布。使用SAS6.12软件进行单因素方差分析。结果:VM-26和CDDP对Tca8113细胞的半数抑制浓度分别为0.35μg/ml和1.1μg/ml。透射电镜观察细胞形态发生典型的凋亡表现,5.0μg/ml和0.15μg/ml的VM-26作用72h,细胞凋亡率分别为81.01%和17.38%,而细胞周期则分别被阻滞在S期和G2/M期。结论:VM-26可以显著诱导口腔鳞癌细胞凋亡并抑制细胞生长,不同剂量的VM-26对口腔鳞癌细胞周期阻滞的阶段不同。

PURPOSE:The present study is designed to identify quantitatively the in vitro effect of VM-26 against oral squamous cell carcinoma Tca8113 and to explore the underlying mechanism. METHODS: Human tongue squamous cell carcinoma cell line Tca8113 was used as subject. Cells were incubated with VM-26 of various concentrations for a variety of time spans. Cisplatin (CDDP) was employed as a control agent. MTT assay was used to assess the inhibition of cell growth by VM-26. Flow cytometer (FCM), transmission electron microscope (TEM) and fluorescence staining were employed for determining the apoptotic rate of cells induced by VM-26. Cell cycle distribution of Tca8113 cells incubated with VM-26 was examined through flow cytometer assay. SAS 6.12 was used for one-way ANOVA. RESULTS: The IC50 of VM-26 against Tca8113 cells was 0.35μg/ml and that of CDDP was 1.1μg/ml. The changes in morphology and ultrastructure of Tca8113 cells were observed using fluorescence microscope and TEM, and apoptotic morphological feature could be found in the nucleus. Apoptotic rate of Tca8113 cells incubated with VM-26 of 5.0μg/ml for 72 hours was 81.01% and cell cycle appeared was arrested at S phase. However, exposed to VM-26 of 0.15μg/ml for 72 hours, Tca8113 cells in the G2/M phase increased from 12.75% to 98.71%, while the apoptotic rate of these cells was 17.38% and far below than that of cells treated with VM-26 of 5.0μg/ml. CONCLUSION: VM-26 could significantly induce apoptosis of oral squamous cell carcinoma cells and inhibit cell growth. There may be another pathway to induce apoptosis of oral squamous cell carcinoma cells except for G2/M phase arrest. Supported by National Natural Science Foundation of China (30300388, 30171014, 30330580) and Research Grant (No.036505013, 034107002) of Science and Technology Committee of Shanghai Municipality.