
脂多糖对人胃癌细胞环氧化酶-2基因启动子活性的影响
网络出版日期: 2012-03-28
Effects of lipopolysaccharide on activity of COX-2 gene promoter of human gastric cancer cells
Online published: 2012-03-28
目的 研究炎症状态下人胃癌细胞中环氧化酶-2(COX-2)基因启动子活性。方法 采用免疫组织化学染色方法观察人胃癌组织中COX-2的表达情况。构建人COX-2基因启动子重组质粒pGL3-COX-2-promoter,采用脂质体介导转染法将重组质粒和内参质粒pRL-TK共转染胃癌细胞株SGC-7901和BGC-823。以不同质量浓度的脂多糖(LPS)工作液刺激细胞,于干预后不同时间点收集细胞,利用双荧光报告系统进行双荧光素酶报告基因活性检测,Western blotting检测COX-2蛋白的表达。结果 免疫组织化学染色观察发现,胃癌癌细胞胞质及癌旁黏膜上皮细胞的胞质中均有COX-2蛋白表达。在相同时间点,SGC-7901和BGC-823内COX-2基因启动子活性随LPS质量浓度的上升而增高(P<0.05);高浓度(10 μg/mL)LPS刺激6 h后COX-2基因启动子活性达到峰值,随后逐渐降低 (P<0.05)。Western blotting检测结果显示:在10 μg/mL LPS干预下,SGC-7901和BGC-823内COX-2蛋白表达随LPS干预时间的延长而呈现上升趋势。结论 炎症刺激可激活人胃癌细胞COX-2基因启动子的表达活性。
徐 苓, 马妍慧, 袁向亮, 等 . 脂多糖对人胃癌细胞环氧化酶-2基因启动子活性的影响[J]. 上海交通大学学报(医学版), 2012 , 32(3) : 283 . DOI: 10.3969/j.issn.1674-8115.2012.03.010
Objective To investigate the activity of cyclooxygenase-2 (COX-2) gene promoter of human gastric cancer cells under inflammatory state. Methods The expression of COX-2 in human gastric cancer tissues was detected with immunohistochemical staining. Recombinant pGL3-COX-2-promoter of human COX-2 gene promoter was constructed, and gastric cancer cell line SGC-7901 and BGC-823 were co-transfected with pGL3-COX-2-promoter and pRL-TK with lipofectin reagent-mediated transfection. Cells were collected after treatment with different mass concentrations of lipopolysaccharide (LPS) for different time, the activity of dual-luciferase reporter gene was determined with dualluciferase reporter system, and the expression of COX-2 protein was detected by Western blotting. Results Immunohistochemical staining indicated that COX-2 protein expressed both in cytoplasm of gastric cancer cells and cytoplasm of para-cancer mucosal epithelial cells. At the same time point, the activity of COX-2 gene promoter of SGC-7901 cells and BGC-823 cells increased with the mass concentrations of LPS (P<0.05). The activity of COX-2 gene promoter reached the peak after treatment with high concentration of LPS (10 μg/mL) for 6 h, and then gradually decreased (P<0.05). Western blotting revealed that the expression of COX-2 protein of SGC-7901 cells and BGC-823 cells increased with the time of treatment with 10 μg/mL LPS. Conclusion Inflammatory stimulation may activate COX-2 gene promoter of human gastric cancer cells.
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