It is known that in many tumors high levels of nm23-H1 correlate with low degree of invasiveness. In addition, transfection of cancer cells

with Nm23-H1 cDNA decreases their metastatic potential. However, the mechanism by which Nm23-H1 suppresses tumor metastasis CCI-779 clinical trial is still poorly understood. Tumor metastasis involves adhesive and migratory events in addition to proteolytic degradation of ECM [6], all of which require the continuous and coordinated formation and disassembly of adhesive structures. It involves stable attachment of a cell to the extracellular matrix at its leading edge which requires transmembrane receptors of the integrin family. Tariquidar in vitro integrins are a super-family, and each of its members is a heterodimer composed of two noncovalently associated different subunits (α and β). At least 14 α and 8 β subunits have been discovered. The sizes of the α subunits are varied between 120~180 kDa, and those of β subunits are

between 90~110 kDa. Most integrins are expressed on the surface of a wide variety of cells, and most cells express several integrins [7]. For example, α5 β1 integrin is a typical receptor of Fn [8] on HepG2 and Hep3B hepatocarcinoma cell lines [9]. ECM-integrin interaction generates intracellular signaling, which induces focal adhesion, actin cytoskeleton formation, cell migration, cell growth, and expression of various genes. To achieve correct cellular function through cell-matrix interaction, the ligation and clustering

of integrins with their ligands need to be regulated in a number of ways. One way is to modulate the expression levels of integrins on cell surface. Another is to AZD6738 supplier regulate the activity of integrins. Hydroxychloroquine concentration It has been indicated that stimulation of β1 integrin by matrix protein initiates intracellular signaling pathways in many types of cells [10–12]. One of the initial events triggered by stimulation of β1 integrin is the association of its cytoplasmic domain with FAK, a cytosolic non-receptor tyrosine kinase, which leads to the tyrosine phosphorylation and activation of FAK [13, 14]. Phosphorylated FAK is involved in the activation of many signal transduction molecules and affects several cellular biological behaviors [10, 11, 14]. In this report, we have studied cell adhesion, spreading and migration, as well as phosphorylation of FAK to fibronectin matrix in H7721 cell line transfected with Nm23-H1 cDNA. Furthermore, the expression of α5 and β1 integrin subunits in H7721 cells was examined, in an attempt to elucidate the molecular mechanism of suppressive effect of Nm23-H1 on cell invasion. Materials and methods Antibodies and Reagents The human hepatocarcinoma H7721 cell line was obtained from the Institute of Cell Biology, Academic Sinica of China. RPMI 1640 and Geneticin (G418) were purchased from Invitrogen. Monoclonal antibody (mAb) of mouse anti-human Nm23-H1 was from Neomarkers Company.