We have discovered that two tomato cystine-knot miniproteins (TCMP) possess in vitro anti-angiogenic activity at sub-micromolar concentrations. The two tomato miniproteins inhibit ERK phosphorylation and HUVEC migration capacity, without altering cell viability and proliferation. To gain a mechanistic insight into the inhibitory action of TCMP on in vitro tube formation, an expression profile analysis of genes involved in human angiogenesis pathway was performed. The majority of the 85 angiogenesis-related genes analysed did not show statistically significant differences in the expression level between EGF- and EGF/TCMP–treated cells. Two genes integrin-alpha V (ITGAV) and Beta-2-microglobulin (B2M) showed a decreased expression in EGF/TCMP–treated HUVEC, whereas vascular endothelial growth factor A (VEGFA) expression was increased.
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