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

摘要

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, ³²P-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 CO₂ 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.