The Study Of Gene Expression Profile And The Preparation Of DNA Microarray Of K562 Cells

Current studies have demonstrated that genetic alterations, either single base mutation or multiple genes comprehensive malfunctioning, will lead to the pathogenesis in the development or disease states. Therefore, the elucidation of gene expression profiling in different circumstances will expand our understanding of the mechanism underlies biological development and physiology, providing further guidance in disease prevention and treatment.K562 is a cell line derived from human erythropoietic leukemia with high malignancy. However, K562 cells maintain certain characteristics that developed into an excellent in vitro model for the study of malignant regulation and cell differentiation. Among these characteristics, certain features are notable, for example, the K562 cells, though possess low differentiation and grow quickly, are susceptible to induced differentiation by various external agents, which lead to 0 -globin expression and the reversion of certain malignant phenotype of the K562 cells.In this study, we applied the techniques of RD桺CR (Restriction Display PCR) to the study of Hemin induced differential gene expression of K562 cells. Four differential expression fragments were isolated and sequenced. One of the genes, after sequencing, turned out to be highly homologous with the cDNA sequence of the ornithine decarboxylase antizyme (OAZ). Furthermore, we applied the RD桺CR technology to amplify grouped cDNA fragments and to construct the K562 cDNA fragments library. Further PCR amplifications of these cDNA clones permit these indexed fragments be used as DNA probes in DNA microarray preparation. DNA microarrays were constructed using Cartesian Microarrayer, and cDNA fragments are arrayed upon microscopic slides. Further study of the K562 gene expression DNA microarrays are underway.In summary, by using an induced differentiation in vitro model and by applying the RD桺CR techniques, we studied hemin induced K562 cells and found that OAZ gene expression may play roles in the phenotype transition. We have also demonstrated that RD 桺CR technology can be a valuable tool in the preparation of considerable amount of cDNA fragments quickly, so as to facilitate the construction of DNA microarrays.