冬凌草甲素联合塞来昔布对人骨肉瘤MG-63细胞增殖及凋亡的影响  

Oridonin in combination with celecoxib inhibits the proliferation and apoptosis of human osteosarcoma MG-63 cells in vitro

作　　者：梁福东 狄淑芳[1] 常云峰[1] 李宏伟[1] 王栓科[1] 

机构地区：[1]兰州大学第二医院骨科,甘肃省骨关节疾病研究重点实验室,甘肃兰州730000

出　　处：《肿瘤》2015年第3期260-267,共8页Tumor

基　　金：甘肃省创新研究群体计划项目(编号:2013GS10047)~~

摘　　要：目的 :检测二萜类化合物冬凌草甲素联合环氧合酶-2(cyclooxygenase-2,Cox-2)抑制剂塞来昔布对人骨肉瘤MG-63细胞增殖及凋亡的影响,并探讨可能的作用机制。方法 :MTT法检测不同浓度的冬凌草甲素(20、40和60μmol/L)和塞来昔布(25、50和100μmol/L)单药和联合处理12和24 h后MG-63细胞的增殖抑制率;FCM法检测冬凌草甲素(40μmol/L)和塞来昔布(50μmol/L)单药和联合干预MG-63细胞后的凋亡率,并分别采用实时荧光定量PCR法和蛋白质印迹法检测对Cox-2以及凋亡调节因子[B细胞性淋巴瘤-2(B cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)和生存素(survivin)]m RNA及蛋白表达水平的影响。结果 :不同浓度的冬凌草甲素和塞来昔布单药和联合干预后,MG-63细胞的增殖明显受到抑制(P值均<0.05),呈剂量依赖性,且在低剂量和中剂量时两药有协同作用。冬凌草甲素和塞来昔布联合用药组较空白对照组和各单药组细胞的凋亡率增加(P值均<0.05)。联合用药组中Cox-2、Bcl-2和survivin m RNA及蛋白的表达水平均较空白对照组和各单药组明显下调(P值均<0.05),而Bax m RNA及蛋白的表达水平明显上调(P值均<0.05)。结论 :冬凌草甲素联合塞来昔布可协同抑制MG-63细胞,这一过程可能与Cox-2的表达受到抑制和凋亡调节因子表达水平的改变有关。Objective： To investigate the effects of diterpenoid oridonin combined with cyclooxygenase-2 （Cox-2） inhibitor celecoxib on proliferation and apoptosis of human osteosarcoma MG-63 cells, and to explore its possible mechanism. Methods： The inhibitory rates of proliferation of MG-63 cells after treatment with different concentrations of oridonin （20, 40 and 60 ~mol/L） and celecoxib （25, 50 and 100 i^mol/L） alone or in combination for different time periods were detected by MTT assay. The apoptosis rates of MG-63 cells after treatment with oridonin （40 μmol/L） and celecoxib （50 μmol/L） alone or in combination were detected by flow cytometry. The expression levels of Cox-2 and apoptosis regulatory factors B cell lymphoma-2 （Bcl-2）, survivin and Bcl-2-associated X protein （Bax） mRNAs and proteins were detected by real-time fluorescence-based quantitative-PCR and Western blotting, respectively. Results： The proliferation inhibition rates of MG-63 cells in single drug groups （oridonin or celecoxib） and the combination group （oridonin and celecoxib） were increased as compared with that in the control group （without any treatment） （P 〈 0.05） in a dose-dependent manner. Oridonin and celecoxib had a synergistic effect at a low or moderate dose. The apoptosis rate of MG-63 cells after treatment with combination of oridonin and celecoxib was higher than those of the control group （without any treatment） and the single drug groups （P 〈 0.05）. The expression levels of Cox-2, Bcl-2 and survivin mRNAs and proteins in combination group were significantly down-regulated as compared with those in the control group （without any treatment） and the single drug groups （all P 〈 0.05）, whereas the expressions of Bax mRNA and protein were up-regulated （all P 〈 0.05）. Conclusion： Oridonin in combination with celecoxib can synergistically inhibit the proliferation of MG-63 cells. This mechanism may be associated with the inhibition of expressions of Cox-2 an

关 键 词：骨肉瘤 抗肿瘤联合化疗方案 细胞凋亡 冬凌草甲素 塞来 昔布 

分 类 号：R738.1[医药卫生—肿瘤]