Low molecular weight heparin inhibits melanoma cell adhesion and migration through a PKCa/JNK signaling pathway inducing actin cytoskeleton changes

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.

► LMWH inhibits M5 and A375 melanoma cell adhesion and migration.
► Protein kinase C alpha (PKCa) phosphorylation is downregulated by LMWH.
► pPKCa downregulation and redistribution to nuclear region attenuates JNK activation.
► JNK inhibition induces M5 cell cytoskeleton changes.
► The lineage activation of PKCa -JNK/p38 regulates M5 cell adhesion/migration.