Analysis of lysophosphatidic acid in metastasis

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Lysophosphatidic acid (LysoPA) has been reported to stimulate mitogenesis of cells and formation of stree figer and/or ruffling. These data suggest that LysoPA has important roles in both cell growth and regulation of actin fibers. On the other hand, LysoPA has been reported to stimulate the metastasis of some cancer cells. First, we analyzed the intracellular signaling of LysoPA.When Swiss 3T3 cells were treated with LysoPA,LysoPA induced the phosphorylation of focal adhesion kinase (FAK) , paxillin, and tensin on tyrosine resides. Next we examinded if phospholipase C (PLC) gamma1 is concerned in the intracellular signaling of LysoPA or not. We made a polyclonal antibody against the phosphotyrosine residues of PLCgamma1 which are phosphorylated by treatment of PDGF.When this antibody was microinjected into the cells, the formation of ruffling by LysoPA was inhibited. Furthermore, tyrosine-phosphorylated PLCgammal was localized in the area of rffling. These results show that PLCgammal exists in the downstream of LysoPA.