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.
-
Key words:
- COX-2 /
- E-cadherin /
- Celecoxib /
- PGE2 /
- Migration /
- Gastric carcinoma
-
-
[1] Jemal A, Bray F, Center MM,et al. Global cancer statistics[J]. CA Cancer J Clin,2011,61(2): 69-90. [2] Carneiro P,Fernandes MS,Figueiredo J,et al.E-cadherin dysfunction in gastric cancer-cellular consequences, clinical applications and open questions[J]. FEBS Lett,2012,586(18): 2981-9. [3] Zheng X.The recent development of the relationship between cyclooxygenase-2 and gastric cancer[J].Xi Nan Jun Yi,2012,14(3):493-6.[郑旭.环氧化酶2（COX-2）与胃癌关系的 研究进展[J].西南军医,2012,14(3):493-6.] [4] Almeida PR, Ferreira FV, Santos CC,et al.Immunoexpression of cyclooxygenase-2 in primary gastric carcinomas and lymph node metastases[J].World J Gastroenterol,2012,18(8): 778-84. [5] Ford AC.Chemoprevention for gastric cancer[J].Best Pract Res Clin Gastroenterol,2011,25(4-5): 581-92. [6] KeXiang Z, YuMin L, Xun L,et al.Study on the association of COX-2 genetic polymorphisms with risk of gastric cancer in high incidence Hexi area of Gansu province in China[J].Mol Biol Rep,2011,38(1): 649-55. [7] Thiel A, Mrena J,Ristimäki A.Cyclooxygenase-2 and gastric cancer[J].Cancer Metastasis Rev,2011,30(3-4): 387-95. [8] Corso G, Marrelli D, Roviello F. Familial gastric cancer and germline mutations of E-cadherin[J]. Ann Ital Chir, 2012,83(3): 17 7-82. [9] Bocca C, Bozzo F, Ievolella M,et al.A novel nitro-oxy substituted analogue of rofecoxib reduces human colon cancer cell growth[J]. Mol Cell Biochem, 2012,361(1-2): 105-10. [10] Shar AO, Gaudard MA, Heath EI,et al.Modeling using baseline characteristics in a small multicenter clinical trial for Barrett's esophagus[J]. Contemp Clin Trials,2009,30(1): 2-7. [11] Sitarz R, Leguit RJ,de Leng WW,et al.Cyclooxygenase-2 mediated regulation of E-cadherin occurs in conventional but not earlyonset gastric cancer cell lines[J].Cell Oncol,2009,31(6): 475-85.
计量
- 文章访问数: 1697
- HTML全文浏览量: 464
- PDF下载量: 846
下载:
