title: Low molecular weight heparin inhibits melanoma cell adhesion and migration through a PKCa/JNK signaling pathway inducing actin cytoskeleton changes.
abstract: Low molecular weight heparin (LMWH) has significant antimetastatic capabilities and affects cancer progression in humans through, not fully defined mechanisms. Here we evaluated its activity at the intracellular level and how it is correlated with melanoma cell adhesion and migration. LMWH inhibited M5 and A375 melanoma cell adhesion and migration in a dose-dependent manner (p⩽0.01). Treatment of M5 melanoma cells with LMWH caused a marked down regulation of constitutive as well as the FN-induced phosphorylation (p⩽0.01) of protein kinase C alpha (PKCa). This was associated with a profound decrease in the cytoplasmic pPKCa (p⩽0.05) and a simultaneous enhancement of nuclear pPKCa localization (p⩽0.01). A significant decrease in the levels of pJNK (p⩽0.01), which is a downstream effector of PKCa, was also demonstrated in the LMWH-treated cells. Furthermore, LMWH-treated cells had disorganized actin stress fibers correlated to a strong decrease in cell-substratum interface area (p⩽0.05) and altered morphology. The decrease in the activation of PKCa, which is an important regulator of cell motility, was directly correlated to the reduced ability of the LMWH-treated melanoma cells to adhere onto and migrate towards the fibronectin (FN) substrate (p⩽0.01). The lineage activation of PKCa-JNK/p38 and their correlation to M5 cell adhesion was confirmed with the utilization of specific inhibitors. In conclusion, LMWH through the downregulation of pPKCa and redistribution to nuclear region attenuates JNK activation, which in turn induces cytoskeleton changes correlated to M5 cell decreased adhesion/migration. This may provide clues for the pharmacological targeting of melanoma.
authors: Department of Histology-Embryology, Medical School, University of Crete, Heraklion, Greece.
authors: Georgia
authors: G
authors: Chalkiadaki
authors: Dragana
authors: D
authors: Nikitovic
authors: Pavlos
authors: P
authors: Katonis
authors: Aikaterini
authors: A
authors: Berdiaki
authors: Aristidis
authors: A
authors: Tsatsakis
authors: Ioanna
authors: I
authors: Kotsikogianni
authors: Nikos K
authors: NK
authors: Karamanos
authors: George N
authors: GN
authors: Tzanakakis
mesh_terms: D000199:Actins / Q000378:metabolism*; D001483:Base Sequence; D002448:Cell Adhesion / Q000187:drug effects*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism*; D017931:DNA Primers; D004789:Enzyme Activation; D006495:Heparin, Low-Molecular-Weight / Q000494:pharmacology*; D006801:Humans; D048670:MAP Kinase Kinase 4 / Q000378:metabolism*; D008545:Melanoma / Q000473:pathology*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D000199:Actins; D017931:DNA Primers; D006495:Heparin, Low-Molecular-Weight; D017868:Cyclic AMP-Dependent Protein Kinases; D048670:MAP Kinase Kinase 4