ABSTRACT
To demonstrate the effect of AB serum on terminal erythroid differentiation ex vivo. Methods: After separation of CD34+ cells from cord blood, the cells were cultured and divided into a control group and an experimental group. The effects of AB serum were examined by the expressions of different markers (GPA, Band3 and α4-integrin) for erythroblast differentiation and enucleation by flow cytometry. Results: The CD34+ cells were successfully differentiated to enucleated red blood cells. There were evident differences among the expressions of GPA, Band3 and α4-integrin between the 2 groups. The percentage of GPA positive cells in the experimental group was bigger than that in the control group in every time point. The expression of Band3 in the experimental group was higher than that in the control group. The expression of α4-integrin in the experimental group was lower than that in the control group. In addition, the enucleation rate in the experimental group was higher than that in the control group. Conclusion: AB serum can promote the cell differentiation and enucleation during terminal erythroid differentiation in vitro.
Subject(s)
Humans , ABO Blood-Group System , Blood , Physiology , Anion Exchange Protein 1, Erythrocyte , Metabolism , Antigens, CD34 , Blood , Cell Differentiation , Genetics , Physiology , Cell Nucleus , Cells, Cultured , Erythrocytes , Physiology , Erythropoiesis , Genetics , Physiology , Fetal Blood , Cell Biology , Physiology , Flow Cytometry , Glycophorins , Metabolism , Integrin alpha4beta1 , MetabolismABSTRACT
OBJECTIVE@#To explore the down-expression mechanism of MYETS1 gene in multiple myeloma cell lines ARH-77 or KM3, and express MYETS1 gene in prokaryotic express system.@*METHODS@#The region of chromosome 13q14.3 in ARH-77 and KM3 was detected by FISH. MYETS1 gene was amplified by RT-PCR and cloned into prokaryotic expression vector pGEX-4T.@*RESULTS@#Positive consequence was acquired in 13q14.3 where MYETS1 located by FISH in ARH- 77 and KM3 cell lines. Bioinformatics indicated highly sequence homology between MYETS1 and LECT1, but excluded the homology of open reading frame between MYETS1 and that of LECT1 by RT-PCR. Myets1 protein was expressed and harvested successfully.@*CONCLUSION@#The region of chromosome 13q14.3 ,where MYETS1 gene located, was not defected in ARH-77 and KM3 cell lines. Down-expression of MYETS1 might be regulated by other mechanisms in multiple myeloma cell lines.
Subject(s)
Humans , Cell Line, Tumor , Chromosomes, Human, Pair 13 , Genetics , Gene Deletion , Genetic Vectors , Genetics , Intercellular Signaling Peptides and Proteins , Genetics , Membrane Proteins , Genetics , Multiple Myeloma , Genetics , Metabolism , Pathology , Neoplasm Proteins , Genetics , Recombinant Proteins , GeneticsABSTRACT
WRKY proteins, a big family of transcription factors, are involved in regulation diverse developmental and physiological processes in plants. Here, a novel WRKY gene, OsWRKY52, was isolated from a rice cDNA library. This gene included an open reading frame of 1 719 bp in length, and the deduced polypeptide contained 572 amino acids,sharing 54% identity with a WRKY1 protein from Avena sativa. Expression of OsWRKY52 gene was induced rapidly by Magnaporthe grisea in the incompatible interaction with rice plant. OsWRKY52 protein, expressed prokaryotically bound specifically to W box cis elements derived from the promoter of a rice PR1a. Transcriptional activation assay was performed by a yeast one- hybrid method. Regions of transactivation were identified to be the N-terminal serine- and threonine-rich islands and the C-terminal acidic domain of OsWRKY52. These results suggest that OsWRKY52, as a transcription activator, may be involved in defense responses against Magnaporthe grisea in rice plants.