Hatanaka Hisashi, Takada Shuji, Choi Young Lim, Fujiwara Shin-ichiro, Soda Manabu, Enomoto Munehiro, Kurashina Kentaro, Watanabe Hideki, Yamashita Yoshihiro, Sugano Kentaro, Mano Hiroyuki
Division of Functional Genomics, Jichi Medical University, Shimotsukeshi, Tochigi 329-0498, Japan.
Biochem Biophys Res Commun. 2007 May 11;356(3):723-6. doi: 10.1016/j.bbrc.2007.03.048. Epub 2007 Mar 19.
Colorectal cancer (CRC) is one of the leading causes of cancer death in humans. In order to identify novel cancer-promoting genes in CRC, we here constructed a retroviral cDNA expression library from a CRC cell line RKO, and used it for a focus formation assay with mouse 3T3 fibroblasts, leading to the identification of 42 independent cDNAs. One of such cDNAs turned out to encode purinergic receptor P2Y, G-protein coupled, 2 (P2RY2). The oncogenic potential of P2RY2 was confirmed in vitro with the focus formation assay as well as soft agar-growth assay, and also in vivo with a tumorigenicity assay in nude mice. While our P2RY2 cDNA encodes a protein with two amino-acid substitutions compared to the reported one, we have confirmed that the wild-type P2RY2 has a strong transforming potential as well. These results indicate an unexpected role of P2RY2 in the carcinogenesis of human cancers.
结直肠癌（CRC）是人类癌症死亡的主要原因之一。为了在结直肠癌中鉴定新的促癌基因，我们在此从结直肠癌细胞系RKO构建了逆转录病毒cDNA表达文库，并将其用于小鼠3T3成纤维细胞的集落形成试验，从而鉴定出42个独立的cDNA。其中一个cDNA编码嘌呤能受体P2Y，G蛋白偶联，2型（P2RY2）。通过集落形成试验以及软琼脂生长试验在体外证实了P2RY2的致癌潜力，并且通过裸鼠致瘤性试验在体内也得到了证实。虽然我们的P2RY2 cDNA编码的蛋白质与报道的蛋白质相比有两个氨基酸替换，但我们也证实野生型P2RY2也具有很强的转化潜力。这些结果表明P2RY2在人类癌症发生中具有意想不到的作用。
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