环氧合酶2抑制剂塞来昔布对结肠癌多药耐药逆转作用的研究  

Reversal effect of cyclooxygenase-2 inhibitor on multi-drug resistance of colon cancer line

作　　者：贺娜[1] 刘春英[1] 宋伟[1] 李芊蔚[1] 

机构地区：[1]大连大学附属中山医院消化科,辽宁大连116001

出　　处：《临床荟萃》2012年第11期950-952,954,F0002,共5页Clinical Focus

摘　　要：目的探讨环氧合酶2(COX-2)抑制剂塞来昔布对结肠癌多药耐药(MDR)的逆转作用。方法在结肠癌多药耐药细胞株colo320/5-FU中,应用四甲基偶氮唑蓝(MTT)方法检测塞来昔布对5-氟尿嘧啶(5-FU)的耐药逆转倍数;反转录-聚合酶链反应(RT-PCR)方法检测耐药基因的变化;免疫组织化学法检测细胞膜跨膜蛋白p-170(P-gp)的表达。结果 MTT结果显示塞来昔布对结肠癌细胞的耐药产生明显的逆转作用(F=2 285.660,P<0.01),其逆转倍数分别为1.70,3.80,8.48,且呈浓度依赖性。RT-PCR结果显示塞来昔布明显抑制多药耐药基因1(MDR1)mRNA的表达(F=986.776,P<0.01),且呈浓度依赖性减低,各浓度组之间差异有统计学意义(P<0.01)。免疫荧光结果显示,细胞膜P-gp荧光强度随着塞来昔布浓度的逐渐增高而减弱。结论选择性COX-2抑制剂塞来昔布对多药耐药细胞株colo320/5-FU具有明显逆转作用,在非细胞毒剂量范围内呈现剂量依赖性,初步机制可能涉及干预多种转录因子的表达而下调MDR1,继而阻滞细胞周期及增加细胞凋亡。Objective To explore the reversal effect of eyclooxygenase-2（COX-2） inhibitor celecoxib on multidrug resistance of colon cancer line. Methods In colon cancer cell lines colo320/5-FU, methyl thiazolyl tetrazolium （MTT） assay was used determine the resistance reversal multiples of celecoxib on five fluorouracil（5-FU）;reverse transcription-polymerase chain reaction （ RT-PCR ） was used to detect the changes of resistance gene; immunohistochemistry was used to detect P-gp expression of cell membrane. Results MTT analysis showed that celecoxib was able to produce significant reversal effect on colon cancer cells in a dose-dependent manner in vitro（ F = 2 285. 660, P 〈0. 001）, and the reversal multiples were 1.70,3.80,8.48 ; Non-cytotoxic dose of celecoxib significantly inhibited multi-drug resistance 1 （MDR）1 mRNA expression in a dose-dependent（ F ： 986. 776, P 〈0.01）. And the inhibition had significant differences between different concentration groups（ P 〈0.0]）. Immunofluorescence showed that P-gp membrane fluorescence intensity gradually weakened as the concentration of celecoxib increased. Conclusion Selective COX-2 inhibitor celecoxih can effectively reverse the colo320/5-FU cells, and that role acts as a dosedependent in non-cytotoxic dose of celecoxib, the initial mechanism may involve intervention of several transcription factors and reduce the expression of MDR1 ,and cause cell cycle arrest and increase apoptosis.

关 键 词：结肠肿瘤 前列腺素内过氧化物合酶类 P糖蛋白 

分 类 号：R735.3[医药卫生—肿瘤]