人结肠癌Caco-2细胞羧酸酯酶的表达及代谢活性  

Expression and metabolic activity of carboxylesterases in human colorectal carcinoma Caco-2 cells

作　　者：刘东[1] 张程亮[1] 杨燕[2] 向道春[1] 高静[1] 王新桃[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院药学部,湖北省武汉市430030 [2]华中科技大学同济医学院附属同济医院实验医学研究中心,湖北省武汉市430030

出　　处：《世界华人消化杂志》2010年第3期294-297,共4页World Chinese Journal of Digestology

摘　　要：目的:评价Caco-2细胞对酯类药物的水解活性及羧酸酯酶(CES)表达的影响.方法:采用人类CES1(hCE-1)和CES2(hCE-2)特异性底物咪达普利和伊立替康(CPT-11)与Caco-2细胞S9组分共同孵育,HPLC法测定原药及代谢产物浓度,评价Caco-2细胞的药物水解活性,并通过RT-PCR法测定细胞内hCE-1和hCE-2表达.结果:Caco-2细胞与正常小肠微粒体酶相比,更倾向于代谢咪达普利,而几乎不水解CPT-11.RT-PCR结果发现,Caco-2细胞CES表达与正常肠道上皮组织不同,表现为hCE-1高表达,而几乎不表达hCE-2.结论:Caco-2细胞CES表达与正常肠道上皮组织存在较大差异,因此在使用Caco-2细胞评价酯类药物口服吸收时应十分谨慎.AIM：To investigate the expression of carboxylesterases（CEs） in human colorectal carcinoma Caco-2 cells and evaluate their catalytic activity for hydrolysis of estercontaining drugs.METHODS：The hydrolytic capacity of Caco-2 cells were evaluated using two substrates, imidapril and irinotecan（CPT-11）, the specific substrates of human CE-1 and human CE-2, respectively.After co-incubation of the two substrates with Caco-2 cell 9 000 g supernatant（S9 fraction）, the two drugs and their metabolic prod-ucts were assayed by high-performance liquid chromatography（HPLC）.Furthermore, the major CE isoforms expressed in Caco-2 cells were detected by reverse transcription-polymerase chain reaction（RT-PCR）.RESULTS：Compared with the human small intestinal microsomes, Caco-2 cells tend to hydrolyze imidapril instead of CPT-11.RT-PCR results indicated that hCE-1 was highly expressed in Caco-2 cells.In contrast, the expression of hCE-2 mRNA was hardly detected in Caco-2 cells.CONCLUSION：The expression pattern of CEs in Caco-2 cells is completely different from that in human small intestine and colon.Therefore, caution should be exercised in predicting human intestinal absorption of ester-containing drugs based on transport results obtained using Caco-2 cells.

关 键 词：羧酸酯酶 CACO-2细胞 伊立替康 咪达普利 

分 类 号：R735.35[医药卫生—肿瘤]