Effects of angiopoietin-1 on attachment and metastasis of human gastric cancer cell line BGC-823

World J Gastroenterol. 2009 Nov 21;15(43):5432-41. doi: 10.3748/wjg.15.5432.

Abstract

Aim: 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).

Methods: 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.

Results: 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).

Conclusion: 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.

MeSH terms

  • Adenoviridae / metabolism
  • Angiopoietin-1 / biosynthesis*
  • Angiopoietin-1 / physiology
  • Carcinoma / metabolism
  • Carcinoma / pathology
  • Cell Adhesion
  • Cell Line, Tumor
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Hyaluronan Receptors / biosynthesis
  • Integrin beta1 / biosynthesis
  • Matrix Metalloproteinase 2 / biosynthesis
  • Neoplasm Invasiveness
  • Neoplasm Metastasis*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stomach Neoplasms / metabolism
  • Stomach Neoplasms / pathology*
  • Urokinase-Type Plasminogen Activator / biosynthesis

Substances

  • Angiopoietin-1
  • CD44v6 antigen
  • Hyaluronan Receptors
  • Integrin beta1
  • Urokinase-Type Plasminogen Activator
  • Matrix Metalloproteinase 2