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.
Virchows Arch. 2010 Dec;457(6):669-76. doi: 10.1007/s00428-010-0960-2. Epub 2010 Oct 3.
Lysophosphatidic acid (LPA), which is a bioactive phospholipid, interacts with specific G protein-coupled transmembrane receptors. Recently, alterations of LPA receptor genes have been reported in some tumor cells. In this study, we examined the expression profiles and DNA methylation status of LPA receptor 1-5 (LPA1-5) genes in human colon cancer cells and also looked for the mutations. Reverse transcription-polymerase chain reaction (PCR) and bisulfite sequencing analyses were carried out. While LPA1, LPA2, and LPA4 genes were expressed in DLD1, SW480, HCT116, CaCo-2, SW48, and LoVo cells, the expressions of LPA3 and LPA5 genes were various. These expression levels were correlated with DNA methylation status in the 5' upstream regions of the LPA receptor genes. Mutation analysis was also performed using a PCR-single-strand conformation polymorphism method. Although no mutations in LPA1, LPA3 and LPA5 genes were found in all types of cells, LPA2 mutations in DLD1 and SW48 cells, and LPA4 mutation were found in DLD1 cells. On the basis of the present results, we demonstrate that these colon cancer cells will be available to understanding the molecular pathway through LPA receptors in the development of tumor cells, and that LPA receptors may be new molecular targets for therapeutic approaches and chemoprevention.
溶血磷脂酸（LPA）是一种具有生物活性的磷脂，与特定的 G 蛋白偶联跨膜受体相互作用。最近，一些肿瘤细胞中 LPA 受体基因的改变已经被报道。在这项研究中，我们检测了人结肠癌细胞中 LPA 受体 1-5（LPA1-5）基因的表达谱和 DNA 甲基化状态，并寻找突变。进行了逆转录-聚合酶链反应（PCR）和亚硫酸氢盐测序分析。虽然 DLD1、SW480、HCT116、CaCo-2、SW48 和 LoVo 细胞中表达了 LPA1、LPA2 和 LPA4 基因，但 LPA3 和 LPA5 基因的表达则不同。这些表达水平与 LPA 受体基因 5'上游区域的 DNA 甲基化状态相关。还使用 PCR-单链构象多态性方法进行了突变分析。尽管在所有类型的细胞中都没有发现 LPA1、LPA3 和 LPA5 基因的突变，但在 DLD1 和 SW48 细胞中发现了 LPA2 突变，在 DLD1 细胞中发现了 LPA4 突变。基于目前的结果，我们证明这些结肠癌细胞将有助于了解肿瘤细胞中通过 LPA 受体发展的分子途径，并且 LPA 受体可能是治疗方法和化学预防的新的分子靶点。
