Obatoclax与MG-132对食管癌细胞CaES-17的协同抗肿瘤作用  

Synergistic anti-tumor effect of obatoclax and MG-132 in esophageal cancer cell line CaES-17

作　　者：赵旭燕[1] 林庆焕 阙富昌 古春萍[1] 余乐[1] 刘叔文[1] 

机构地区：[1]南方医科大学药学院,广东广州510515

出　　处：《南方医科大学学报》2016年第4期506-513,共8页Journal of Southern Medical University

基　　金：国家自然科学基金(81273551)~~

摘　　要：目的探讨obatoclax联用MG-132对人食管癌细胞是否具有协同的抗肿瘤作用。方法 MTT法检测obatoclax与MG-132对人食管癌细胞Ca ES-17的细胞毒作用,根据IC50确定两药联用的浓度比(1∶2.4),将两个药物从4 IC50的作用浓度起倍比稀释,用Compu Syn软件计算两药的联用指数(CI)。采用Western blot方法研究药物对ubiquitin表达、PARP蛋白剪切、caspase-9蛋白剪切、phospho-Histone H3蛋白及phospho-Aurora A/B/C蛋白表达的影响。采用流式细胞术检测细胞早期凋亡(Annexin-V阳性率)及细胞周期的影响。结果 Obatoclax与MG-132联合处理人食管癌细胞Ca ES-17,对细胞抑制率为50%及95%时相对应的CI值分别为0.296及0.104,具有明显的协同抗肿瘤作用。单用obatoclax或MG-132均能诱导ubiquitin表达增加、PARP及caspase-9发生剪切,且联用组与单用组相比有统计学差异(P<0.05);联用组phospho-Histone H3蛋白增加与单用组相比有统计学差异(P<0.05);phospho-Aurora A/B/C蛋白表达的增加也与单用obatoclax相比有统计学差异(P<0.05),与单用MG-132相比无明显改变(P>0.05)。同时,联用组Annexin-V阳性率及sub-G1期及G2/M期细胞百分比与单用组相比都显著增加,且差异均有统计学意义(P<0.05)。结论 Obatoclax联用MG-132对人食管癌细胞Ca ES-17具有协同抗肿瘤作用,且与诱导肿瘤细胞发生凋亡及G2/M期阻滞有关。Objective To explore whether MG-132 could enhance the anti-tumor activity of obatoclax against esophageal cancer cell line CaES-17. Methods MTT assay was used to determine the cytotoxicity of obatoclax and MG-132 in CaES-17 cells. The IC50 of obatoclax and MG-132 were used to determine the molar ratio （1∶2.4） of the two drugs for combined treatment of the cells. The concentrations of obatoclax and MG-132 ranged from 1/8 IC50 to 4 IC50 after serial dilution, and their combination index （CI） was calculated using CompuSyn software. The expression of ubiquitin and the cleavage of PARP, caspase-9, phospho-histone H3 and phospho-aurora A/B/C in the exposed cells were examined with Western blotting;the cell apoptosis was measured by flow cytometry with Annexin V staining, and the percentage of cells in each cell cycle phase was also determined by flow cytometry. Results The CI of obatoclax and MG-132 was 0.296 for a 50%inhibition of Caes-17 cells and was 0.104 for a 95%inhibition. The cells treated with obatoclax or MG-132 alone showed increased expression of ubiquitin and cleavage of PARP and caspase-9. Compared with the cells treated with obatoclax or MG-132 alone, the cells with a combined treatment exhibited significantly increased expression of ubiquitin, cleavage of PARP and caspase-9, and expression of phospho-Histone H3 （P〈0.05）. The combined treatment of the cells also resulted in significantly increased expression of phospho-Aurora A/B/C compared with obatoclax treatment alone. The cells with the combined treatment showed significantly higher percentages of apoptotic cells and cells in sub-G1 and G2/M phases compared with the cells treated with either of the drugs （P〈0.05）. Conclusion Obatoclax combined with MG-132 shows a significant synergistic anti-tumor effect against esophageal cancer CaES-17 cells by inducing apoptosis and cell cycle arrest.

关 键 词：食管癌 MG-132 协同抗肿瘤作用 细胞凋亡 周期阻滞 

分 类 号：R735.1[医药卫生—肿瘤]