COX-2表达下调抑制人结肠癌细胞系HT-29增殖并促进凋亡  

Down-regulation of COX-2 expression inhibits proliferation and promotes apoptosis in human colon cancer cell line HT-29

作　　者：刘海云[1] 钟睿[2] 谢琛[2] 王侠 何志坚[2] 

机构地区：[1]江西中医药大学基础医学院,江西南昌330004 [2]江西省肿瘤医院放疗四科,江西南昌330029

出　　处：《基础医学与临床》2017年第11期1568-1573,共6页Basic and Clinical Medicine

基　　金：江西省自然科学基金(20142BAB205109)

摘　　要：目的探讨环氧化酶-2(COX-2)表达对大肠癌细胞HT-29细胞的细胞活力、增殖和凋亡等生物学行为的影响。方法利用脂质体将siRNA COX-2及空载体转入HT-29大肠癌细胞中,并设立对照组。48 h后,real-time PCR法检测COX-2、Bcl-2、Bax和基质金属蛋白酶2(MMP2)mRNA表达;Western blot检测COX-2及p-AKT表达;CCK-8法检测细胞活力;Annexin V/PI流式双染检测细胞凋亡;流式细胞计量术检测细胞周期;划痕实验及Transwell实验分别检测细胞迁移和侵袭能力。结果与对照组及空载体组比较,COX-2抑制组COX-2蛋白及mRNA表达量显著降低(P<0.01);细胞活力下降(P<0.01);细胞凋亡率提高,细胞周期阻滞于G期,细胞迁移及侵袭能力降低(P<0.01);Bcl-2及MMP2 mRNA表达下调(P<0.01);Bax mRNA表达上调(P<0.01);p-AKT蛋白表达下调(P<0.01)。结论 COX-2表达量下调后能显著的抑制细胞增殖、迁移与侵袭,并诱导细胞凋亡,可能与下调p-AKT有关。Objective To explore effect of cyclooxygenase-2（COX-2）expression on cell viability,proliferation,apoptosis and other biological behavior of human colon cancer cell line HT-29.Methods The expression of COX-2was examed by Western blot and real-time PCR after siRNA COX-2,empty body was transferred into human colorectal cancer cell line HT-29 by liposome.Cell viability was measured by CCK-8 method.Cell apoptosis and cell cycle was measured by flow cytometry.Cell migration and invasion was detected by scratch test and Transwell method,respectively.The expression of Bcl-2,Bax,matrix metalloproteinase 2（MMP2）was measured by real-time PCR.The level of protein kinase B（AKT）phosphorylation was measured by western blot.Results Compared with the control group and empty vector group,the expression of COX-2 protein and mRNA was decreased（P〈0.01）,cell viability was reduced（P〈0.01）,cell apoptotic rate was increased（P〈0.01）,cell cycle was arrested in G phase（P〈0.01）,cell migration and invasion ability was decreased（P〈0.01）,the expression of Bcl-2,MMP2 mRNA and p-AKT protein was down-regulated（P〈0.01）,the expression of Bax mRNA was up-regulated in COX-2 inhibition group（P〈0.01）.Conclusions Down-regulation of COX-2 expression can significantly inhibit HT-29 cell proliferation,migration and invasion,and induce cell apoptosis,which may be related to the down-regulation of AKT phosphorylation.

关 键 词：环氧化酶-2(COX-2) 大肠癌 HT-29 生物学行为 

分 类 号：R735.35[医药卫生—肿瘤]