Fax +41 61 306 12 34 E-Mail karger@karger.ch www.karger.com Original Paper Pathobiology 2010;77:278–282 DOI: 10.1159/000319875 Mutations of Lysophosphatidic Acid Receptor Genes in Human Osteosarcoma Cells Kyoko Okabe   a Mai Hayashi   a Minako Fujii   a Kanya Honoki   c Toshio Mori   d Nobuyuki Fukushima   b Toshifumi Tsujiuchi   a   Divisions of a  Cancer Biology and Bioinformatics, and b  Molecular Neurobiology, Department of Life Science, Faculty of Science and Engineering, Kinki University, Higashiosaka, c  Department of Orthopedic Surgery, and d  RI Center, Nara Medical University, Kashihara, Japan LPA3. Conclusion: These results indicated that the mutations in LPA1 and LPA3 genes occur in MG63 cells, suggesting that the alterations in LPA receptor genes may play some role in the pathogenesis in human osteosarcoma cells. Copyright © 2010 S. Karger AG, Basel Introduction Lysophosphatidic acid (LPA) is a bioactive mediator and induces several cellular effects, including regulation of cell proliferation, differentiation, transcellular migra- tion, morphogenesis and protection from apoptosis [1–6]. LPA can induce cell proliferation, migration, invasion and production of angiogenic factors in human ovarian cancer cell lines, suggesting that LPA may play an impor- tant role in the development of tumor cells [2, 3, 7]. LPA interacts with at least 6 G-protein-coupled transmem- brane receptors, LPA receptor-1 (LPA1), LPA2, LPA3, LPA4, LPA5 and LPA6 [8–12]. Previously, aberrant ex- pressions of LPA receptors have been reported in human ovarian, colorectal and thyroid tumors, indicating that alterations in LPA receptor gene expression might also be Key Words Lysophosphatidic acid  Lysophosphatidic acid receptor  Mutation  Sarcoma Abstract Objective: Lysophosphatidic acid (LPA), which is a bioactive phospholipid, interacts with specific G protein-coupled trans- membrane receptors. Recently, alterations in LPA receptor genes have been reported in some tumor cells. In this study, to assess an involvement of LPA receptor genes in the devel- opment of human cancer cells, we looked for the presence of mutations in LPA receptor 1–6 (LPA1–6) genes in MG63 osteo- sarcoma, HT1080 fibrosarcoma, A549 lung adenocarcinoma, MCF-7 breast carcinoma, and G-361 melanoma cells. Meth- ods: Genomic DNAs were extracted from each cell and poly- merase chain reaction-single-strand conformation polymor- phism analysis was performed to identify the mutations. Results: MG63 showed mutations in LPA1 and LPA3 genes, while no mutations in the LPA receptor genes were found in HT1080, A549, MCF-7 and G-361 cells. Sequence analysis re- vealed a CGC to CGT (Arg to Arg) transition at codon 314 in LPA1, and a GCG to GTG (Ala to Val) transition at codon 247 in Received: May 25, 2010 Accepted after revision: July 27, 2010 Toshifumi Tsujiuchi Laboratory 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) Tel. +81 6 6772 2332, Fax +81 6 6723 2721, E-Mail ttujiuch  @  life.kindai.ac.jp © 2010 S. Karger AG, Basel 1015–2008/10/0775–0278$26.00/0 Accessible online at: www.karger.com/pat