阻滞真核细胞起始因子-4E对膀胱癌细胞乙酰肝素酶表达的影响  

Effect of Eukaryotic initiation factor-4E inhibition on heparanase expression in bladder cancer

作　　者：庄锦涛[1] 涂响安[1] 王文卫[1] 赵良运[1] 赵亮[1] 丘少鹏[1] 邓春华[1] 胡斌[2] 

机构地区：[1]中山大学附属第一医院黄埔院区泌尿外科,广州510700 [2]中山大学达安基因诊断中心

出　　处：《中华实验外科杂志》2012年第5期921-923,共3页Chinese Journal of Experimental Surgery

基　　金：广东省省部产学研结合项目（20118090400034）;广东省自然科学基金资助项目（9151802904000002）;广东省科技计划资助项目（2008803031082）;广东省自然科学基金博士启动项目（7301521）;广州市黄埔区科技攻关课题（0732）

摘　　要：目的探讨膀胱癌细胞真核细胞起始因子-4E（eIF4E）与乙酰肝素酶表达的关系。方法脂质体包裹eIF-4E于mRNA翻译起始点互补的反义寡核苷酸（asODN）及相应对照的正义寡核苷酸（sODN）以100、200pmol／L两种浓度分别转染膀胱癌EJ细胞，逆转录-聚合酶链反应（RT—PCR）检测eIF4E转录水平的改变。荧光定量PCR和Westernblot分别检测乙酰肝素酶mRNA及蛋白表达。结果asODN转染显著抑制eIF4E基因水平。eIF-4E被阻滞后，乙酰肝素酶mRNA水平及蛋白表达量：asODNl组（0．276±0．050、0．255±0．028）及asODN2组（0．195±0．022、0．160±0．032），均显著低于空白对照组（0．896±0．057、0．603±0．029）及sODN1组（0．867±0．099、0．572±0．029）和sODN2组（0．879±0．087、0．560±0．035，P〈0．01）。结论asODN转染膀胱癌EJ细胞可以阻滞eIF-4E基因表达，阻滞eIF-4E后，膀胱癌乙酰肝素酶mRNA水平及蛋白表达均明显降低。Objective To investigate whether eukaryotic initiation factor-4E （eIF-4E） in human bladder cancer cell may alter the expression levels of hcparanase. Methods Antisense oligodeoxynucleoti- de （asODN） targeted against the translation start site of eIF-4E mRNA and sense oligodeoxynucleotide （sODN） with the concentration of t00,200 pmol/L was introduced into EJ cells by means of liposome-me- diated DNA-transfeetion. elF-4E mRNA levels were tested by reverse transcription-polymerase chain reac- tion （RT-PCR）. Fluorescence quantitative RT-PCR and Western blotting were used to test the expression levels of heparanase mRNA and protein respectively. Results AsODN against eIF-4E significantly inhibi- ted eIF-4E gene expression. Following cIF-4E inhibition, expression levels of heparanase mRNA and pro-tein treated with asODN1 was 0. 276±0. 050 and 0. 255±0. 028, treated with asODN2 was 0. 195 ±0. 022 and 0. 160± 0. 032, both significantly less than 0. 896±0. 057 and 0. 603±0. 029 in control group, 0. 867±. 099 and 0. 572±0. 029 in sODN1 group, 0. 879±0. 087 and 0. 560 ± 0. 035 in sODN2 group （ P 〈 0. 01 ）. Conclusion AsODN transfected into human bladder cancer cell EJ can significantly inhibited eIF-4E gene expression. And the inhibition of eIF-4E reduces the expression of heparanase mRNA and protein.

关 键 词：膀胱癌 真核细胞起始因子4E 乙酰肝素酶 反义寡核苷酸 转染 

分 类 号：R737.14[医药卫生—肿瘤]