Bevacizumab modulates the process of fibrosis in vitro

Clin Exp Ophthalmol. 2015 Mar;43(2):173-9. doi: 10.1111/ceo.12374. Epub 2014 Aug 19.


Background: Fibrosis is the most common side effect after anti-vascular epithelial growth factor (VEGF) therapy (intravitreal bevacizumab) for retinal or choroidal neovascularization. This study was to investigate the efficacy of bevacizumab on the expressions of fibrosis-related cytokines in human umbilical vein endothelial cells (HUVECs) in vitro.

Methods: Cultured HUVECs were divided into groups of controls (group 1), hypoxia (group 2) and hypoxia combined with bevacizumab (group 3). No treatment was given in group 1. In group 2, cobalt(II) chloride (CoCl₂) (200 μm) was added to the medium. In group 3, in addition to CoCl₂, bevacizumab was mixed in the medium, with a final concentration of 0.25 mg/mL, roughly equal to the concentration used clinically. The expressions of connective tissue growth factor (CTGF), transforming growth factor-β₂ (TGF-β₂) and basic fibroblast growth factor-2 (bFGF-2) were evaluated by SYBR green real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay at 6 h, 12 h, 24 h and 48 h. Matrix metalloproteinases (MMP)-2 was detected by SYBR green real-time PCR and Western blotting at each time point.

Results: Both messenger RNA and protein levels of CTGF, bFGF, TGF-β₂ and MMP-2 in group 2 were higher than group 1 (P < 0.05). In group 3, the expressions of CTGF, bFGF, TGF-β₂ and MMP-2 were upregulated compared with group 2 (P < 0.05).

Conclusions: Bevacizumab at clinical doses can exert pro-fibrotic effects on HUVECs by upregulating the expressions of CTGF, bFGF, TGF-β₂ and MMP-2. This may be involved in fibrosis after anti-VEGF therapy.

Keywords: anti-VEGF; bevacizumab; cytokines; fibrosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiogenesis Inhibitors / pharmacology*
  • Antibodies, Monoclonal, Humanized / pharmacology*
  • Antimutagenic Agents / toxicity
  • Bevacizumab
  • Blotting, Western
  • Cell Hypoxia
  • Cells, Cultured
  • Cobalt / toxicity
  • Connective Tissue Growth Factor / genetics
  • Connective Tissue Growth Factor / metabolism
  • Cytokines / genetics*
  • Cytokines / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Fibroblast Growth Factor 2 / genetics
  • Fibroblast Growth Factor 2 / metabolism
  • Fibrosis
  • Human Umbilical Vein Endothelial Cells / drug effects*
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Human Umbilical Vein Endothelial Cells / pathology
  • Humans
  • Matrix Metalloproteinase 2 / genetics
  • Matrix Metalloproteinase 2 / metabolism
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Retinal Vessels / pathology*
  • Transforming Growth Factor beta2 / genetics
  • Transforming Growth Factor beta2 / metabolism
  • Vascular Endothelial Growth Factor A / antagonists & inhibitors


  • Angiogenesis Inhibitors
  • Antibodies, Monoclonal, Humanized
  • Antimutagenic Agents
  • CCN2 protein, human
  • Cytokines
  • RNA, Messenger
  • Transforming Growth Factor beta2
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • Fibroblast Growth Factor 2
  • Connective Tissue Growth Factor
  • Bevacizumab
  • Cobalt
  • Matrix Metalloproteinase 2
  • cobaltous chloride