Department of Gastroenterology, Zhongda Hospital, Southeast University, Nanjing 210009, Jiangsu Province, China.
World J Gastroenterol. 2009 Nov 21;15(43):5432-41. doi: 10.3748/wjg.15.5432.
To evaluate the effects of angiopoietin-1 (Ang-1) on adhesion of gastric cancer cell line BGC-823 and expression of integrin beta1, CD44V6, urokinase-type plasminogen activator (uPA) and matrix metalloproteinase-2 (MMP-2).
BGC-823 cells were transfected transiently with adenovirus-Ang-1 (Ad-Ang-1). Cells transfected transiently with adenovirus-green fluorescent protein (Ad-GFP) and untransfected cells were used as a negative and blank control group, respectively. The cell adhesion rate between cell and extracellular matrix (ECM) was determined by cell adhesion assay. To investigate whether Ang-1 could reinforce gastric carcinoma metastasis, we performed migration and invasion assays in BGC-823 cells. The mRNA and protein expression of integrin beta1, CD44V6, uPA and MMP-2 were detected by reverse transcription polymerase chain reaction and Western blotting, respectively. The expression of integrin beta1 and CD44V6 was measured by immunohistochemistry.
BGC-823 cells were transfected successfully. The adhesion rate increased significantly in the Ad-Ang-1 group (P < 0.05). The Ad-Ang-1-transfected group had a significant increase in migration and invasion compared with that of the mock-transfected and Ad-GFP groups. The mRNA and protein expression of integrin beta1, CD44V6, uPA and MMP-2 in the Ad-Ang-1 group was higher than that in the Ad-GFP and blank control groups (P < 0.05). Compared with mock-transfected and Ad-GFP groups, integrin beta1 and CD44V6 expression intensity greatly increased (P < 0.05).
Transfection of Ang-1 into human gastric cancer cell line BGC-823 can significantly increase expression of integrin beta1 and CD44V6, by which cell adhesion and metastasis to the ECM are promoted.
评价血管生成素-1（Ang-1）对胃癌细胞系 BGC-823 黏附及整合素β1、CD44V6、尿激酶型纤溶酶原激活物（uPA）和基质金属蛋白酶-2（MMP-2）表达的影响。
瞬时转染 BGC-823 细胞腺病毒-Ang-1（Ad-Ang-1）。瞬时转染腺病毒-绿色荧光蛋白（Ad-GFP）和未转染的细胞分别作为阴性和空白对照组。通过细胞黏附试验测定细胞与细胞外基质（ECM）之间的黏附率。为了研究 Ang-1 是否可以增强胃癌转移，我们在 BGC-823 细胞中进行了迁移和侵袭试验。通过逆转录聚合酶链反应和 Western 印迹法分别检测整合素β1、CD44V6、uPA 和 MMP-2 的 mRNA 和蛋白表达，通过免疫组织化学法检测整合素β1 和 CD44V6 的表达。
BGC-823 细胞转染成功。Ad-Ang-1 组细胞黏附率显著增加（P<0.05）。与 mock 转染组和 Ad-GFP 组相比，Ad-Ang-1 转染组的迁移和侵袭能力显著增强。Ad-Ang-1 组整合素β1、CD44V6、uPA 和 MMP-2 的 mRNA 和蛋白表达均高于 Ad-GFP 组和空白对照组（P<0.05）。与 mock 转染组和 Ad-GFP 组相比，整合素β1 和 CD44V6 的表达强度显著增加（P<0.05）。
转染 Ang-1 到 BGC-823 人胃癌细胞系可显著增加整合素β1 和 CD44V6 的表达，从而促进细胞与 ECM 的黏附和转移。
