Fukui Rie, Tanabe Eriko, Kitayoshi Misaho, Yoshikawa Kyohei, Fukushima Nobuyuki, Tsujiuchi Toshifumi
Division of Cancer Biology and Bioinformatics, Department of Life Science, Faculty of Science and Engineering, Kinki University, 3-4-1 Kowakae, Higashiosaka, Osaka 577-8502, Japan.
Tumour Biol. 2012 Dec;33(6):1899-905. doi: 10.1007/s13277-012-0450-z. Epub 2012 Jul 5.
Lysophosphatidic acid (LPA) mediates a wide range of biological responses with G protein-coupled transmembrane receptors (LPA receptors). So far, at least six types of LPA receptors (LPA receptor-1 (LPA(1)) to LPA(6)) have been identified. Recently, it has been reported that LPA(3) indicates opposite effects on cellular functions of cancer cells. In the present study, to assess a biological role of LPA(3) on cell migration ability of colon cancer cells, we generated LPA receptor-3 (LPAR3) knockdown (HCT-sh3-3) cells from HCT116 and measured cell motile and invasion activities. In motility assay with a cell culture insert, HCT-sh3-3 cells showed significantly high cell motile activity, compared with control cells. For invasion assay, the filter was coated with Matrigel. The invasive activity of HCT-sh3-3 cells was significantly higher than that of control cells. Furthermore, we also examined the effects of LPAR3 knockdown on the interaction between colon cancer cells and endothelial F-2 cells. When F-2 cells were cultured with serum-free DMEM containing a supernatant from HCT-sh3-3 cells, the cell growth rate and migration activity of F-2 cells were significantly stimulated, associating with the elevated expressions of vascular endothelial growth factor (VEGF)-A and VEGF-C genes in HCT-sh3-3 cells. These results suggest that LPA(3) may act as a negative regulator on cell motile and invasive abilities of colon cancer HCT116 cells.
溶血磷脂酸（LPA）通过G蛋白偶联跨膜受体（LPA受体）介导多种生物学反应。到目前为止，至少已鉴定出六种类型的LPA受体（LPA受体-1（LPA(1)）至LPA(6)）。最近，有报道称LPA(3)对癌细胞的细胞功能具有相反的作用。在本研究中，为了评估LPA(3)对结肠癌细胞迁移能力的生物学作用，我们从HCT116细胞中生成了LPA受体-3（LPAR3）敲低（HCT-sh3-3）细胞，并测量了细胞的运动和侵袭活性。在使用细胞培养插入物的运动性测定中，与对照细胞相比，HCT-sh3-3细胞表现出明显更高的细胞运动活性。对于侵袭测定，滤膜用基质胶包被。HCT-sh3-3细胞的侵袭活性明显高于对照细胞。此外，我们还研究了LPAR3敲低对结肠癌细胞与内皮F-2细胞之间相互作用的影响。当F-2细胞与含有HCT-sh3-3细胞上清液的无血清DMEM一起培养时,F-2细胞的细胞生长速率和迁移活性受到显著刺激，这与HCT-sh3-3细胞中血管内皮生长因子（VEGF）-A和VEGF-C基因表达的升高有关。这些结果表明，LPA(3)可能对结肠癌HCT116细胞的运动和侵袭能力起负调节作用。
