Effects of thalidomide on the expression and secretion of vascular endothelial growth factor and tumor necrosis factor-alpha by a human keratinocyte cell line HaCaT / 中华皮肤科杂志

ABSTRACT

Objective To investigate the effect of thalidomide on the proliferation of as well as the expression and secretion of vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α) by the human keratinocyte cell line HaCaT.Methods Cultured HaCaT cells were divided into several groups to be treated with dimethyl sulfoxide (negative control group) and various concentrations (0.01nmol/L-100 μmol/L) of thalidomide (experimental groups) respectively for 20 to 24 hours.Subsequently,water soluble tetrazolium-1(WST-1) assay was performed to estimate cellular proliferation,real time quantitative PCR to detect the mRNA expression of VEGF and TNF-α in HaCaT cells,and enzyme-linked immunosorbent assay (ELISA) to quantify the protein expressions of VEGF and TNF-α in the culture supernatants of HaCaT cells.Statistical analysis was done by one-way analysis of variance with least significant difference post hoc test.Results The survival rate of HaCaT cells was 74.3％,82.9％ and 90.8％ after 24-hour treatment with thalidomide of 100,10 and 1 μmol/L respectively,significantly lower than that in the negative control group (100％,all P ＜0.01).A significant decrease was induced in the mRNA expression (0.439-to 0.634-fold change,all P ＜0.01) and supematant level ((0.587-to 0.923-fold change,P ＜0.05) of VEGF in HaCaT cells by thalidomide of 0.01-100 nmol/L,as well as in the mRNA expression (0.493-to 0.587-fold change,P ＜0.05) and supernatant level (0.408-to 0.617-fold change,P ＜0.01) of TNF-α by thalidomide of 0.1-100 nmol/L.Conclusion Within a certain range of concentration,thalidomide could suppress the proliferation of,as well as the expression and secretion of VEGF and TNF-α by,HaCaT cells.