Studies On The Changes Of Gene Expression During The Chemical-induced Erythroid Differentiation Of Murine Erythroleukemia Cells (MEL) By MRNA Differential Display

Erythropoiesis is one of the most important aspects in cell differentiation, during which the expression of the erythroid-related genes prevails, and the balance between proliferation, apoptosis and differentiation of the different-developmental-stage erythroid cells is maintained. What are involved in this complex course? Answering this question will help us to elucidate the molecular mechanism of erythroid differentiation, and allow a theoretical expounding of the causes of related blood diseases and their therapeutics. Hence, searching and identifying the erythroid-differentiation-related genes is an important subject in the erythropoiesis research. Our experimental objective is to investigate the differential expression of genes during the chemical-induced erythropoiesis, and to screen the erythroid-differentiation-associated genes.We choose a classical cell models—the Murine Erythroleukemia Cells (MEL) as experimental material, and adopt the mRNA differential display technique to analyze the changes of the gene expression during the DMSO and Hemin co-induction of erythroid differentiation.Firstly, MEL cells are treated for 5 days with DMSO and Hemin, and assays for cell differnetiation by the Benzidine staining method. After harvesting induced and non-induced cells, we isolate the total cellular RNAs and synthesize the 1st and 2nd strand of cDNA. Then the pseudo-adapter are ligated to the ends of the Rsal-digested double strand cDNAs fragments, and T primer and Ad primer (which corresponds to the outer region of adapter long strand) are used for the basic amplification of the ligation production to congregate cDNA fragments containing the oligo (A/T).