Wakabayashi Naoko, Tsujino Megumu, Tajiri Masaki, Taki Midori, Koshino Ayumi, Ikeda Hiroko, Fukushima Nobuyuki, Tsujiuchi Toshifumi
J Toxicol Pathol. 2010 Mar;23(1):63-6. doi: 10.1293/tox.23.63. Epub 2010 Apr 5.
Lysophosphatidic acid (LPA) is a bioactive phospholipid that stimulates cell proliferation and migration, and protects cells from apoptosis. It interacts with specific G protein-coupled transmembrane receptors. Recently, frequent mutations of the LPA receptor-1 (LPA1) gene were detected in rat lung adenocarcinomas induced by N-nitrosobis(2-hydroxypropyl)amine (BHP). In this study, to evaluate the involvement of other LPA receptor gene alterations during lung carcinogenesis, we investigated mutations of the LPA2, LPA3, LPA4 and LPA5 genes in lung adenocarcinomas induced by BHP in rats. Fifteen male Wistar rats, 6 weeks of age, were given 2000 ppm BHP in their drinking water for 12 weeks and then maintained without further treatment until sacrifice at 25 weeks, and 15 adenocarcinomas were obtained. Genomic DNAs were extracted from frozen tissues, and the LPA2, LPA3, LPA4 and LPA5 genes were examined for mutations, using polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) analysis. No mutations of LPA2, LPA3, LPA4 and LPA5 were detected in the 15 adenocarcinomas. These results suggest that alterations due to LPA2, LPA3, LPA4 and LPA5 gene mutations might not be involved in the development of lung adenocarcinomas induced by BHP in rats.
溶血磷脂酸（LPA）是一种生物活性磷脂，可刺激细胞增殖和迁移，并保护细胞免于凋亡。它与特定的G蛋白偶联跨膜受体相互作用。最近，在N-亚硝基双（2-羟丙基）胺（BHP）诱导的大鼠肺腺癌中检测到LPA受体-1（LPA1）基因频繁突变。在本研究中，为了评估其他LPA受体基因改变在肺癌发生过程中的作用，我们调查了BHP诱导的大鼠肺腺癌中LPA2、LPA3、LPA4和LPA5基因的突变情况。15只6周龄的雄性Wistar大鼠，在其饮用水中给予2000 ppm BHP，持续12周，然后在不进行进一步处理的情况下维持，直至25周时处死，获得了15个腺癌样本。从冷冻组织中提取基因组DNA，使用聚合酶链反应（PCR）-单链构象多态性（SSCP）分析检测LPA2、LPA3、LPA4和LPA5基因的突变情况。在这15个腺癌样本中未检测到LPA2、LPA3、LPA4和LPA5的突变。这些结果表明，LPA2、LPA3、LPA4和LPA5基因突变引起的改变可能与BHP诱导的大鼠肺腺癌的发生无关。
