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Isolating Bronchial Epithelial Cells from Resected Lung Tissue for Biobanking and Establishing Well-Differentiated Air-Liquid Interface Cultures
Published on: May 26, 2023
[Toluene diisocyanate increases vascular endothelial growth factor expression in human bronchial epithelial cells]
Hong-juan Peng1, Shao-xi Cai, Hai-jin Zhao
1Department of Respiratory Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. lisiyiabc@tom.com
Objective:
To investigate the effect of toluene diisocyanate (TDI) on the expression of vascular endothelial growth factor (VEGF) in human bronchial epithelial (HBE) cells.
Methods:
TDI-human serum albumin (TDI- HSA) conjugate was prepared using a modified Son's method. MTT assay was used to examine the viability of HBE135-E6E7 cells cultured in serum-free medium after treatment with HSA or TDI-HSA at different concentrations. VEGF mRNA expression of the HBE cells treated with HSA or TDI-HSA at 10, 20, 30 and 40 microg/ ml, respectively, was detected using semi-quantitative RT-PCR.
Results:
Treatment with 40 microg/ml HSA and 40 microg/ml TDI-HSA did not result in significant changes in the viability of HBE135-E6E7 cells. RT-PCR revealed the constitutive expression of two VEGF isoforms, namely VEGF189 and VEGF165, in cultured HBE135-E6E7 cells. After exposure to TDI-HSA at the different concentrations (except for 10 microg/ml), a significant increase occurred in both VEGF189 and VEGF165 mRNA expressions in HBE135-E6E7 cells as compared with the expressions in the control group and the HSA-treated cells (P<0.05), and significant dose dependence was noted in the effect of TDI-HSA (P<0.05). No significant difference was found in the expressions between the control cells and the HAS-treated cells (P>0.05).
Conclusion:
TDI induces significant increase in VEGF expression in HBE cells, and VEGF overexpression may play an important role in the pathogenesis of TDI-induced asthma.

