二苯并呫吨类化合物CY-B12体外抗肿瘤细胞增殖的作用及其机制的初步研究  

In Vitro Inhibitory Effect of Dibenzoxanthene CY-B12 on Proliferation of Tumor Cell Lines and Its Possible Mechanisms

作　　者：蒲兴祥[1,2] 江高峰[1,3] 陈泳[4] 许遵乐[4] 冼励坚[1,3] 

机构地区：[1]华南肿瘤学国家重点实验室 [2]中山大学肿瘤防治中心,广东广州510060 [3]中山大学肿瘤防治中心,广东广州510060 [4]中山大学化学与化学工程学院

出　　处：《癌症》2005年第12期1442-1447,共6页Chinese Journal of Cancer

摘　　要：背景与目的:CY-B12是最近合成的新型二苯并呫吨类化合物,本研究探讨CY-B12的体外抗肿瘤细胞增殖作用及其机制。方法:MTT法测定CY-B12体外抑制胃癌细胞MGC803、鼻咽癌细胞CNE-2、口腔上皮癌细胞KB-3-1及肺癌细胞Glc82等细胞株增殖的作用,流式细胞仪检测CY-B12作用后细胞周期的改变及细胞凋亡,Hoechst33258染色后荧光显微镜观察细胞凋亡形态,Westernblot测定细胞周期相关蛋白Cdc25C、CyclinB1、Cdc2的改变,单细胞凝胶电泳检测CY-B12对细胞DNA的损伤作用。结果:CY-B12对MGC803、CNE-2、KB-3-1及GLC82等细胞株的增殖都有明显的抑制作用,IC50值分别为7.51、9.58、8.84和15.99μmol/L。CY-B12可诱导CNE-2细胞周期阻滞于G2/M期继而发生细胞凋亡,可观察到细胞染色质凝集、凋亡小体产生及凋亡峰出现。CY-B12作用24h后,细胞周期相关蛋白Cdc25C表达水平随CY-B12浓度升高而下调;CyclinB1蛋白在CY-B12浓度低时升高,CY-B12浓度高时表达下降;Cdc2蛋白变化趋势与CyclinB1相同。单细胞凝胶电泳证明CY-B12诱导了DNA损伤,并存在浓度-效应依赖关系。结论:CY-B12有较强的体外抗增殖作用,它可能通过断裂细胞DNA、下调周期相关蛋白Cdc25C、诱导细胞发生周期阻滞和凋亡而发挥其作用。BACKGROUND ＆ OBJECTIVE： CY-B12, a new dibenzon- xanthene, has been synthesized recently. This study was to investigate the in vitro antiproliferative activity of CY-B12 and its possible mechanisms. METHODS： The inhibitory effects of CY-B12 on proliferation of gastric carcinoma cell line MGC803, nasopharyngeal carcinoma cell line CNE-2, oral epithelial carcinoma cell line KB-3-1, and lung cancer cell line Gic82 were assessed by MTT assay. Cell cycle distribution and apoptosis were detected by flow cytometry （FCM）. Apoptotic morphology of CNE-2 cells was observed under fluorescent microscope after Hoechst33258 staining. The expression of cell cycle-related proteins Cdc25C, Cdc2, and Cyclin B1 were measured by Western blot. DNA damage was detected by single cell gel electrophoresis （SCGE）. RESULTS： CY-B12 obviously inhibited the proliferation of MGC803, CNE-2, KB-3-1, and Gic82 cells; the IC50 values were 7.51, 9.58, 8.84, and 15.99 μmol/L, respectively. After treatment of CY-B12, CNE-2 cells were arrested at G2/M phase; chromatin condensation, apoptotic bodies, and sub-G1 peak were observed in CNE-2 cells. The protein expression of Cdc25C in CNE-2 cells was down-regulated by CY-B12 in a dose-dependent manner; whereas Cyclin B1 and Cdc2 were upregulated by low dose of CY-B12, and down-regulated by high dose of CY-B12. CY-B12 induced DNA damage in CNE-2 cells in a dose-dependent manner. CONCLUSION： CY-B12 has potent in vitro antiproliferative activity, which may be exerted through breaking DNA, down-regulating cell cyclerelated protein Cdc25C, and inducing cell cycle arrest and apoptosis.

关 键 词：抗肿瘤药 二苯并呫吨/药理学 CY-B12/药理学 肿瘤细胞株 体外实验 

分 类 号：R73-36[医药卫生—肿瘤]