Analysis of lysophosphatidic acid in metastasis

溶血磷脂酸(LysoPA)可刺激细胞有丝分裂和张力和/或褶皱的形成。这些数据表明，LysoPA在细胞生长和肌动蛋白纤维的调节中都起着重要的作用。另一方面，据报道，LysoPA可以刺激一些癌细胞的转移。首先，我们分析了LysoPA的细胞内信号转导。当LysoPA处理瑞士3T3细胞时，LysoPA诱导粘着斑激酶(FAK)、巴西林和张力蛋白在酪氨酸残基上的磷酸化。接下来，我们研究了磷脂酶C(PLC)Gamma1是否参与了LysoPA的细胞内信号转导。我们制备了一种针对PLCGamma1的磷酸酪氨酸残基的多克隆抗体，该抗体经PDGF处理后被磷酸化，当该抗体显微注射到细胞中时，LysoPA的褶皱形成被抑制。此外，酪氨酸磷酸化的PLC-Gammal也定位于RRAFSING区。这些结果表明，PLC-Gammal存在于LysoPA下游。

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.