Transforming activity of purinergic receptor P2Y, G protein coupled, 8 revealed by retroviral expression screening
Transforming activity of purinergic receptor P2Y, G protein coupled, 8 revealed by retroviral expression screening
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DOI：
10.1080/10428190701225882
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发表时间：
2007-01-01
影响因子：
2.6
通讯作者：
Mano, Hiroyuki
中科院分区：
文献类型：
--
作者：
Fujiwara, Shin-Ichiro;Yamashita, Yoshihiro;Mano, Hiroyuki
双表型急性白血病（BAL）是一种相对罕见的急性白血病亚型，其特征是同时存在髓系和淋巴细胞表面抗原。我们现在已经筛选了转化基因的BAL胚细胞与使用的焦点形成试验与逆转录病毒cDNA表达文库构建从BAL患者分离的恶性胚细胞。从转化灶中回收的一些逆转录病毒插入片段被发现编码野生型嘌呤能受体P2 Y，G蛋白偶联，8（P2 RY 8）。P2 RY 8的致癌潜力用体外病灶形成测定以及用裸鼠体内致瘤性测定来证实。多种基于内切酶的报告基因分析显示，P2 RY 8增加CREB和Elk-1的反式激活活性以及血清应答元件和c-Fos和c-Myc基因的增强子-启动子片段的转录活性。骨髓CD_（34）（+）细胞P_2RY_8mRNA的定量分析表明，白血病患者尤其是难治性白血病患者P_2RY_8mRNA表达增高。因此，我们的数据揭示了P2 RY 8在白血病细胞中的丰富表达及其在急性白血病发病机制中的意想不到的作用。
Biphenotypic acute leukemia ( BAL) is a relatively rare subtype of acute leukemia characterized by the presence of both myeloid and lymphoid cell surface antigens. We have now screened for transforming genes in BAL blasts with the use of the focus formation assay with a retroviral cDNA expression library constructed from malignant blasts isolated from a BAL patient. Some of the retroviral inserts recovered from transformed foci were found to encode wild-type purinergic receptor P2Y, G protein coupled, 8 ( P2RY8). The oncogenic potential of P2RY8 was confirmed with the in vitro focus formation assay as well as with an in vivo tumorigenicity assay in nude mice. A variety of luciferase-based reporter assays revealed that P2RY8 increased both the trans-activation activities of CREB and Elk-1 as well as the transcriptional activities of the serum response element and enhancer-promoter fragments of the c-Fos and c-Myc genes. Quantitation of P2RY8 mRNA in CD34(+) cells of bone marrow showed that P2RY8 expression is frequently increased in leukemia patients, especially in those with refractory disease. Our data thus reveal an abundant expression of P2RY8 in leukemic cells and its unexpected role in the pathogenesis of acute leukemia.