环氧合酶-2抑制剂塞来昔布诱导人肝癌细胞凋亡及其机制  

COX-2 inhibitor celecoxib induces apoptosis of hepatoma carcinoma SMMC-7721 cell line and its mechanism

作　　者：赵军艳[1] 李轩[1] 谢立群[1] 郑艳敏[1] 

机构地区：[1]中国人民武装警察部队医学院附属医院消化科,天津300162

出　　处：《中国肿瘤生物治疗杂志》2010年第3期322-326,共5页Chinese Journal of Cancer Biotherapy

基　　金：武警医学院博士启动基金课题(No.WBS200814)~~

摘　　要：目的:探讨环氧合酶-2(cyclooxygenase-2,COX-2)抑制剂塞来昔布(celecoxib)诱导人肝癌细胞系SMMC-7721凋亡的作用及其可能机制。方法:以10、25、50、75、100μmol/L的塞来昔布处理SMMC-7721细胞,MTT法检测肿瘤细胞增殖,Ho-echst 33342/PI双染法检测细胞凋亡的形态特征,流式细胞术检测细胞凋亡率和细胞周期变化。RT-PCR法检测Fas和Bcl-2mRNA的表达,Western blotting检测细胞Fas和Bcl-2蛋白的表达。结果:塞来昔布以剂量依赖的方式抑制SMMC-7721细胞增殖。塞来昔布作用后,SMMC-7721细胞出现核染色质凝集、核膜破裂等凋亡细胞典型的形态特征。25、50和100μmol/L塞来昔布诱导SMMC-7721细胞的凋亡率分别为(16.32±2.32)%、(38.05±2.47)%、(71.17±3.19)%,并使细胞阻滞于G0/G1期。塞来昔布处理后,SMMC-7721细胞FasmRNA及蛋白表达明显增强(P<0.01);Bcl-2mRNA表达无明显变化(P>0.05),高浓度塞来昔布可下调Bcl-2蛋白表达(P<0.01)。结论:塞来昔布能够抑制SMMC-7721细胞增殖并诱导凋亡,其机制可能与影响凋亡相关基因表达有关。Objective To investigate the effect of COX-2 inhibitor celecoxib on induction of apoptosis of hepatoma carcinoma SMMC-7721 cell line and the possible mechanism.Methods： SMMC-7721 cells were treated with 10,25,50,75 and 100 μmol/L celecoxib;their proliferation was examined by MTT assay;typical apoptotic morphology was studied by Hoechst 33342/PI double staining;and cell cycle and apoptosis rate were detected by flow cytometry.Fas and Bcl-2 mRNA expressions were examined by RT-PCR,and their protein expressions were detected by Western blotting analysis.Results： Celecoxib dose-dependently inhibited the growth of SMMC-7721 cells.SMMC-7721 cells showed typical apoptotic morphology features,such as nuclear chromatin condensation and nuclear membrane rupture,after celecoxib treatment.Apoptotic rates of SMMC-7721 cells treated with 25,50,and 100 μmol/L celecoxib were（16.32±2.32）%,（38.05±2.47）%,and（71.17±3.19）%,respectively;celecoxib induced cell cycle arrest of SMMC-7721 cells in G0/G1 phase.Fas mRNA and protein expressions in celecoxib-treated SMMC-7721 cells were higher than those in the untreated cells（P〈0.01）;the expression of Bcl-2 mRNA was not affected by celecoxib stimulation（P〉0.05）,but Bcl-2 protein expression was significantly down-regulated（P〈0.01）.Conclusion：Celecoxib can inhibit the proliferation and induce apoptosis of SMMC-7721 cells,possibly by regulating the expression of apoptosis related genes.

关 键 词：环氧合酶-2(COX-2) 塞来昔布 肝肿瘤 SMMC-7721细胞 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤] R730.54[医药卫生—临床医学]