siRNA介导COX-2基因沉默对大肠癌LoVo细胞侵袭及上皮间质转化的影响  

Effect of silencing cyclooxygenase-2 by siRNA on inducing epithelial-mesenchymal transition and invasiveness in colon cancer LoVo cells

作　　者：李明[1] 谭诗云[1] LIMing;TAN Shiyun(Department of Gastroenterology,Renmin Hospital of Wuhan University,Hubei Key Laborary of Digestive System Disease,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院消化内科消化系疾病湖北省重点实验室,湖北武汉430060

出　　处：《胃肠病学和肝病学杂志》2018年第12期1363-1366,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：中央高校基本科研业务费专项基金(2042017kf0092)

摘　　要：目的探讨靶向沉默环氧合酶-2(cyclooxygenase-2,COX-2)表达对大肠癌LoVo细胞侵袭能力及上皮间质转化的影响。方法在体外COX-2 siRNA转染大肠癌LoVo细胞,RT-PCR及免疫印迹法验证转染后COX-2 mRNA及蛋白表达,Transwell小室法检测细胞侵袭能力的改变;免疫印迹法检测EMT标志物(E-cadherin、Vimentin、Fibronectin)、ERK/MAPK和PI3K/Akt通路相关蛋白(Akt、p-Akt、p-GSK-3β、ERK、p-ERK)及MMP-9表达的变化情况。结果 COX-2 siRNA转染大肠癌LoVo细胞后,COX-2 mRNA及蛋白表达下调,同时细胞侵袭能力降低;上皮标记物E-cadherin表达上调,同时间叶标记物Vimentin和Fibronectin表达下调;细胞总的ERK、Akt蛋白表达水平无明显改变,而p-ERK、p-Akt表达降低,同时,Akt下游效应蛋白p-GSK-3β表达也下调; MMP-9表达下调。结论 COX-2 siRNA转染有效抑制了大肠癌LoVo细胞COX-2表达,并可能通过阻滞细胞ERK/MAPK和PI3K/Akt通路,进而抑制EMT,下调MMP-9表达,降低细胞侵袭能力。Objective To investigate the effect of silencing cyclooxygenase-2(COX-2) by small interfering RNA(siRNA) on the invasiveness and inducing epithelial-mesenchymal transition in colon cancer LoVo cells.Methods The colon cancer LoVo cells were cultured in vitro.After transfection with COX-2 siRNA,the invasion of LoVo cells was tested by transwell assay.Western blotting was used to detect the expressions of E-cadherin,Vimentin,Fibronectin,MMP-9,Akt,p-Akt,p-GSK-3β,ERK and p-ERK proteins.Results After transfection with COX-2 siRNA,the invasion of LoVo cells was decreased.Silencing COX-2 by siRNA could suppress the expressions of Vimentin,Fibronectin,MMP-9,p-Akt,p-GSK-3β and p-ERK proteins,and promote the expression of E-cadherin.Meanwhile,the expressions of p-Akt and p-ERK were not changed significantly.Conclusion Silencing COX-2 by siRNA could suppress capacity of cell invasion,and the effect may be associated with the blocking of the ERK/MAPK and PI3K/Akt signaling pathways,thus inhibiting the EMT and the expression of MMP-9.

关 键 词：大肠癌 小干扰RNA 环氧合酶-2 上皮间质转化 侵袭 

分 类 号：R735.34[医药卫生—肿瘤]