Overexpression of homeobox gene HOXD3 induces coordinate expression of metastasis-related genes in human lung cancer cells

Homeobox-containing genes are expressed in spatiotemporal fashion during em bryogenesis and act as master transcription-regulating factors which contro l the expression of a variety of genes involved in morphogenesis. They are also expressed in a tissue-specific manner in normal adult tissues and appe ar to give cells spatial information in the maintenance of their architectu ral integrity. We transfected a HOXD3 class I homeobox-containing gene into human lung cancer A549 cells and investigated alterations in gene expressi ons and phenotypes related to the maintenance of tissue architecture in HOX D3-overexpressing A549 cells. In the HOXD3-overexpressing cell lines, expre ssion of E-cadherin was lost and plakoglobin was strongly repressed, wherea s integrin alpha3 and beta3 were up-regulated and N-cadherin and integrin a lpha4 were newly expressed. Compared with parental and control transfectant lines, the HOXD3-overexpressing cell lines showed highly motile and invasi ve activity. Blocking experiments using anti-integrin beta1 and beta3 sugge sted that the increased haptotaxis of the HOXD3-overexpressing cells to vit ronectin resulted from increased expression and activation of integrin alph av beta3, and that overexpression of the HOXD3 gene converted the integrin beta1-dependent haptotaxis to fibronectin into both integrin beta1- and bet a3-dependent one. HOXD3 overexpression increased production of matrix-degra tive enzymes including matrix metalloproteinase-2 and urokinase-plasminogen activator. When the tumor cells were intravenously injected into the tail veins of nude mice, HOXD3 transfectants formed a significantly large number of metastatic foci in lungs compared with the control transfectants. These findings suggest that HOXD3 can act as a metastasis-promoting gene in huma n lung cancer A549 cells, (C) 2001 Wiley-Liss. Inc.