阻抑eIF-4E对大肠腺癌LS-174T细胞乙酰肝素酶mRNA及其翻译表达水平的影响  

Contribution of eIF-4E inhibition to heparanase expression at both mRNA and protein levels in human colon adenocarcinoma cell line LS-174T

作　　者：杨玉捷[1] 钟世顺[1] 简燕婷[1] 万田漠[1] 傅思武[1] 李旭[1] 张亚历[1] 

机构地区：[1]第一军医大学南方医院消化系疾病研究所,广东广州510515

出　　处：《中国病理生理杂志》2004年第10期1895-1899,共5页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目 (No .0 10 6 4 3) ;国家骨干教师基金资助项目

摘　　要：目的 :确定阻抑结肠癌LS - 174T细胞中过量表达的真核细胞起始因子 - 4E(eukaryoticinitiationfactor- 4E ,eIF - 4E)是否促进乙酰肝素酶 (heparanase)mRNA的降解 ,并改变其翻译表达水平。方法 :应用脂质体包裹与eIF - 4EmRNA翻泽起始点互补的asODN ,转染处理人大肠腺癌细胞LS - 174T。使用Westernblot和RT -PCR方法分别检测eIF - 4E被阻抑后其转录和翻译水平的改变。乙酰肝素酶mRNA在细胞内水平采用Northernblot定量检测 ,其蛋白表达水平改变采用Westernblot检测。结果 :asODN经脂质体转染LS - 174T细胞后 ,eIF - 4E基因表达明显受到抑制 ,其蛋白表达产物也显著下降。伴随eIF - 4E被阻抑表达 ,Northernblot结果显示乙酰肝素酶mRNA水平下降 ,且其蛋白翻译表达量也降低。结论 :阻抑eIF - 4E影响LS - 174T细胞乙酰肝素酶mRNA稳定、促使其降解 。AIM: To determine whether the eukaryotic initiation factor-4E (eIF-4E) inhibition facilitates the degradation of heparanase mRNA and alters heparanase protein expression in human colon adenocarcinoma cell line, LS-174T. METHODS: A 20-mer antisense s-oligodeoxynucleotide (asODN) targeted against the translation start site of eIF-4E mRNA were introduced into LS-174T cells by lipid-mediated DNA-transfection. eIF-4E protein and mRNA levels were detected by Western blot and RT-PCR, respectively. The mRNA levels of heparanase were determined by Northern blot. The alterations of heparanase expression were confirmed by Western blot analysis. RESULTS: The 20-mer asODN against eIF-4E specifically and significantly inhibited eIF-4E protein expression. Following eIF-4E inhibition, a significant reduction of heparanase mRNA was observed on Northern blot, and at the same time, heparanase protein expression significantly decreased as well. CONCLUSIONS: The results indicate that the inhibition of eIF-4E strongly reduces the stability of heparanase mRNA in colon adenocarcinoma cell line, LS-174T and resultes in an apparent reduction in the expression of heparanase protein. [

关 键 词：结肠肿瘤 真核细胞起始因子-4E 肝素裂解酶 LS-174T细胞 

分 类 号：R363[医药卫生—病理学]