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环氧合酶-2对人SGC-7901胃癌细胞E-cadherin的表达及迁移能力的影响

Effects of Cyclooxygenase-2 on E-cadherin Expression and Migration of Human Gastric Carcinoma Cell Line SGC-7901 in vitro

  • 摘要: 目的 研究环氧合酶-2(COX-2)通过调控E-钙黏素(E-cadherin)对胃癌细胞侵袭迁移能力的影响。方法 分别应用塞来昔布(Celecoxib)及前列腺素E2(PGE2)对体外培养的人胃癌细胞SGC-7901进行干预,采用实时荧光定量反转录PCR检测COX-2、E-cadherin mRNA的表达;运用免疫荧光标记法结合激光共聚焦荧光显微镜分析E-cadherin的蛋白表达量;应用Transwell法检测细胞迁移能力的变化。结果 实时荧光定量反转录PCR检测塞来昔布明显抑制体外培养人胃癌细胞SGC-7901 中COX-2 mRNA的表达(P<0.01),而E-cadherin mRNA的表达随着COX-2的表达下降而呈浓度与时间依赖性升高(P<0.01);运用PGE2干预后E-cadherin mRNA表达呈浓度与时间依赖性下降(P <0.05或P<0.01)。塞来昔布30 μmol/L干预人胃癌细胞SGC-7901 24、36、48 h后,激光共聚焦荧光显微镜检测E-cadherin的蛋白表达量明显升高(P<0.05);PGE2 1 μmol/L干预24、48 h后,E-cadherin 蛋白表达量明显下降(P<0.05)。塞来昔布组穿过Transwell小室的细胞数明显少于对照组(P <0.01),PGE2组穿过Transwell小室的细胞数明显高于对照组(P<0.05)。结论 COX-2特异性抑制塞来昔布通过抑制COX-2的表达,上调E-cadherin表达量,抑制体外培养人胃癌细胞SGC-7901的迁移能力;外源性PGE2能够下调SGC-7901胃癌细胞E-cadherin表达量从而促进肿瘤细胞的迁移能力。

     

    Abstract: Objective To investigate the effects of cyclooxygenase-2(COX-2) regulating E-cadherin expression on migration capability of gastric carcinoma cell line SGC-7901. Methods After SGC-7901 cell line treated by Celecoxib and PGE2 in vitro, real-time quantitative PCR was used to detect the changes of mRNA levels of COX-2 and E-cadherin. The combination of immunofluorescence and Confocal laser scanning microscopy was used to analyze the protein level of E-cadherin. Transwell was used to detect the change of cell migration. Results COX-2 mRNA expression was inhibited by Celecxib, but E-cadherin mRNA expression level was increased signifi cantly with decreased COX-2 expression, in a dose- and timedependent manner(P<0.01).On the contrary, E-cadherin mRNA expression level was decreased in a timeand dose-dependent manner after PGE2 treatment(P<0.05 or P<0.01). After SGC-7901 cell line were treated with Celecxib (30 μmol/L) for 24,36,48 h, E-cadherin protein level was increased significantly(P<0.05), while E-cadherin protein expression was signifi cantly decreased after PGE2 treatment(1 μmol/L) for 24 and 48 h (P<0.05).The cell numbers through Transwell with Celecoxib treatment was less than that of no-treatment group(P<0.01).While, the cell numbers through Transwell with PGE2 treatment was more than that of notreatment group (P<0.05). Conclusion Celecoxib, selective COX-2 inhibitor, may upregulate E-cadherin expression and inhibit the invasive potential of human gastric carcinoma by suppressing COX-2 expression. PGE2 may downregulate E-cadherin expression and promote the migration of SGC-7901 in vitro.

     

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