The effects of TNF-alpha on ADAMTS-1 and VEGF gene expressions in HEP3B cells

Abstract

Angiogenesis is essential for tumor growth, progression andmetastasis. Angiogenesis is controlled by several endogenousstimulators and inhibitors. Among these stimulators, the vascularendothelial growth factor (VEGF) owns a private value becauseof its importance and specificity in angiogenesis. An importantinhibitor of the angiogenesis is the first described member of theADAMTS (a disintegrin and metalloproteinase with thrombo-spondin) family, ADAMTS-1, and it is demonstrated that AD-AMTS-1 binds and sequestrates VEGF and this results with thesuppression of endothelial cell proliferation.Several factors may change the expression levels of ADAMTS-1and VEGF. This study aims to evaluate the effects of TNF-aonADAMTS-1 and VEGF gene expressions in Hep3B cells in a doseand time dependent manner.Hep3B cells were treated with different doses of TNF-afor dif-ferent time intervals. Total cellular RNAs were prepared from allgroups and subjected to RT-PCR using specific ADAMTS-1 andVEGF primers. GAPDH amplification was carried out as a posi-tive control. Amplification products were analyzed on agarose gelelectroforesis and densitometric analysis were carried out.