华蟾素诱导人膀胱癌细胞株T24凋亡研究

Cinobufacin induced apoptosis in bladder carcinoma cell line T24

目的研究华蟾素注射液（cinobufacin，Cino）对膀胱癌细胞株T24的体外增殖抑制及诱导凋亡作用，探讨其对膀胱癌的l临床应用价值。方法不同浓度Cino单独作用和配伍丝裂霉素C（MMC）作用于膀膀癌T24细胞株后，通过流式细胞术（FCM）以及DNA凝胶电泳检测细胞的凋亡情况，应用光学显微镜观察细胞形态学的变化。结果除Cino（25mg／m1）组外，其它处理组均可看到较为典型的细胞凋亡的形态学变化。DNA琼脂糖电泳可见到凋亡的特征性“梯子”条带。流式细胞仪结果示，除高浓度的Cino（25mg／m1）组外．其余各组均见到在G0／G1期前出现亚二倍体峰（APO峰），且阻滞细胞生长在G0／G1期。各药物浓度组与对照组的S期比率（SPF）和增殖指数（PI）分别进行比较（P〈O．05）有统计学差异。结论Cino对膀胱肿瘤1、24细胞有诱导凋亡和直接杀死的双重作用。具体表现为高浓度（25mg／m1）作用于细胞时，对细胞表现为直接杀死作用。而在较低浓度范围内（O．001～2．5mg/ml），表现为对细胞诱导凋亡的作用，且具有一定的时效和量效关系：配伍丝裂霉素应用时．对T24细胞的诱导凋亡作用明显增强．两药有协同作用。

Objective To explore the value of cinobufacin injection on clinical application, by investigating the effect of cinobufacin on inhibiting proliferation and inducting apoptosis in bladder carcinoma cell T24 in vitro. Methods Apoptosis and proliferation in bladder carcinoma cell line T24 induced by cinobufacin was investigated with flow cytometry （FCM）and DNA agarose gel electrophoresis, when T-24 cell had been cultured with different concentration of cinobufacini injection and MMC, the morphological changes were observed through the optical microscope. Results In addition to Cino （25mg/ml） group, the morphological changes of typical apoptosis were observed by optical microscope in other groups. Agarose gel electrophoresis of DNA revealed the apoptosis characteristic of ＂Ladder＂. Flow cytometry results showed, in addition to high concentrations of Cino （25mg/ml） group, the rest of the groups were seen the hypodiploid peak （APO peak） before the G0/G1 phase, and block cell growth in the G0/G1 phase. The S-phase fraction （SPF） and proliferation index （PI） of each drug group compare to the con- trol group,s, statistically significant difference existed （,~〈0.05）. Conclusion Cinobufaein has dual-action to bladder carcinoma cell T24, induce apoptosis and direct killing . It specifically showed a effect of direct killing in high concentrations （25mg/ml） on the celt. In the lower concentration range （0.001-2.5mg/ml）, showed the role of cell apoptosis, And has a certain time-effect and dose-effect relationship. When Mitomycin（MMC） application compatibility on the T24 cells ,it markedly enhanced induction of apoptosis. Cinobufaein injection have synergistic effect with MMC.With a potential value of clinical application of bladder carcinoma.