环氧化酶2抑制剂对结肠癌细胞中BCL-3及细胞周期蛋白D1表达的影响  

Effect of COX-2 inhibitor on the expression of BCL-3 and cyclin D1 in human colon cancer cell line SW480

作　　者：王少敏[1] 叶孟[1] 倪曙民[1] 吴骁[2] 杨光[2] 

机构地区：[1]宁波大学医学院附属医院肿瘤科,315020 [2]宁波大学医学院附属医院血液科,315020

出　　处：《中华胃肠外科杂志》2010年第8期612-615,共4页Chinese Journal of Gastrointestinal Surgery

基　　金：宁波市社会发展科研项目（2007C10085）

摘　　要：目的 探讨环氧化酶2（COX-2）抑制剂对BCL-3基因及其靶向基因细胞周期蛋白1（cyclin D1）的转录和表达的影响.方法 实验组分别以25、50、100及200 μmol/L的NS398（COX-2抑制剂）处理结肠癌SW480细胞48 h或72 h;对照组以不含NS398的溶媒处理SW480细胞.采用RT-PCR、Western blot及免疫细胞化学染色方法分别从mRNA和蛋白水平检测各组细胞中BCL-3和cyclin D1的表达情况.结果 各组细胞处理48 h后,与对照组相比,实验组BCL-3和cyclin D1 mRNA水平下降,呈剂量依赖性;而蛋白水平则无明显差异（P〉0.05）.处理72 h后,与对照组相比,实验组BCL-3和cyclin D1蛋白水平下降,亦呈现剂量依赖性.当NS398浓度达到100 μmol/L时,其BCL-3 mRNA及蛋白表达水平与对照组比较差异即有统计学意义（P〈0.01）：当NS398浓度达到50 μmol/L时,其cyclin D1 mRNA及蛋白表达水平与对照组比较差异即有统计学意义（P〈0.05）.结论 结肠癌细胞株SW480中有BCL-3的表达.COX-2抑制剂NS398可从基因及蛋白水平对BCL-3及cyclinD1进行抑制,并呈剂量依赖性.NS398可能通过BCL-3而抑制cyclinD1.Objective To study the effects of NS398, a selective cyclooxygenase-2 （COX-2） inhibitor, on the transcription and translation of BCL-3 and its regulatory gene cyclin D1 in colon cancer cell line SW480. Methods Human colon cancer cells SW480 were divided into two groups： SW480 cells in experimental group were treated with NS398 in different concentrations （25 μmol/L, 50 μmol/L, 100 μmol/L and 200 μmol/L） for 48 h or 72 h. SW480 cells in control group were treated with media which did not contain NS398. Then the expressions of BCL-3 and cyclin D1 were detected by RT-PCR, Western blot, and immunocytochemistry. Results At 48 hours RT-PCR showed that BCL-3 mRNA and cyclin D1 mRNA decreased in a dose-dependent manner in the experimental group. However, there were no significant differences in the levels of BCL-3 protein and cyclin D1 protein between two groups （P〉0.05）. At 72 hours, BCL-3 protein and cyclin D1 protein also decreased in a dose-dependent manner in the experimental group. When the concentration of NS398 reached 100 μmol/L, the differences between the two groups in the expression of BCL-3 mRNA and protein became statistically significant （P〈0.01）. When the concentration of NS398 reached 50 μmol/L, the differences in the expression of cyclin Dl mRNA and protein were statistically significant （P〈0.05）. Conclusions BCL-3 is expressed in colon cancer cell line SW480. COX-2 inhibitor can inhibit the expression of BCL-3 and cyclin Dl in a dose-dependent manner. NS398 may down-regulate the expression of cyclin D1 through BCL-3.

关 键 词：结肠肿瘤 环氧化酶2抑制剂 基因 BCL-3 细胞周期蛋白D1 SW480细胞 

分 类 号：R735[医药卫生—肿瘤]