人细胞中Quox-1基因的定位、表达和多态性

MAPPING,EXPRESSION AND RFLP OF QUOX-1 GENE IN HUMAN CELL

用Southern杂交法研究Quox 1基因在人正常白细胞、LCE细胞及其HeLa细胞中的限制性酶切片段多态性 ;免疫组织化学方法研究Quox 1基因在人细胞中的表达。结果表明 :在正常人细胞DNA中存在鹌鹑Quox 1基因的同源序列 ,在LCE等肿瘤细胞中Quox 1基因表现出明显的限制性酶切片段多态性和基因扩增现象。Quox 1蛋白在HeLa、LCE等肿瘤细胞中激活表达 ,而在正常人白细胞未检测出Quox 1基因的表达。Quox 1基因同人类的肿瘤发生、发展和癌变具有一定的相关性。

In order to study the relationship between the Quox 1 and the tumorigenesis and cell maligimization, cellular genomic DNA of normal human leukocytes and some tumor cells (LCE and HeLa cells) were extracted using proteinase K method and were restricted with EcoR Ⅰ. After electrophoresis, and transferred to NC membrane, 32 P labeled Quox 1 cDNA was used as probe to detect the presence of Quox 1 homologous in genomic DNA of normal human, and cancer cells. Normal human leukocyte and some tumor cells (LCE and HeLa cells) were cultured respectively on glass slide with CO 2 incubator. Using antibodies of Quox 1, the expression of Quox 1 protein in Normal human leukocyte, and some tumor cell was detected (LCE cell, HeLa cell). There was homologous sequence of Quox 1 in normal human leukocyte, and some tumor cells (LCE and HeLa cells), it appears Restriction Fragment Length Polymorphism (RFLP) and may undergo gene amplification. In LCE cells, the gene amplification was much greater than that in HeLa cells. The expression of Quox 1 protein could not be detected both in cell nucleus and cytoplasm of normal human leukocyte. But it was activated and expressed in LCE and HeLa cells. However, in the cell nucleus of LCE and Hela cells,Quox 1 gene had strong expression. There was weak expression of Quox 1 protein even in the cytoplasm. The results suggest that the Quox 1 gene may be related with the tumorigenesis and cell aligimization.