The Molecular Mechanism Of Cybridization Regulates Cell Differentiation

Cybridization model was established by Xue Shepu to study erythroid differentiation,dedifferentiation,and regulation of cancer cells by cytoplasmic factor.This model can be used to study the theory and application and cure of malignancy myeloma and neoplastic hematologic disorder.So far the studies of erythroid differentiation have a long history.But an application of proteomics technology on erythroid differentiation is infancy.In this paper,it is application of proteomics technique and the cybridization model hoping to find an important regulative differentiation factor of erythroid differentiation as well as malignant myeloma,neoplastic hematologic disorder and treatment of some clues.The neo and GFP gene(from pEGFP-C1 plasmid DNA)is used as markers;the lipofectin syetem was used to transfer the neo gene and GFP DNA to mouse reticulocytes.K562 cells were then mixed with the reticulocytes at a ratio of 1:20.PEG was employed to promote cell fusion,and cells selected through G418 medium(600ug/ml)to obtain the resistant strain of cybrid cells MR.k562-Rneo.Clonal growth of the cybrid cells emerged on the backgrongd at about the 20th-30th day of culture.In order to examine whether the cybrid cells was induced to recover differentiation,several genes have been tested.These genes are erythroid differentiation related gene,including gaa-1,nf-e2and ?-globin,and cell cycle related gene,including p53 and p16,proliferation related gene,apoptosis related gene,bcl-2,bax,As well as the oncogene,h-ras,and c-myc.Out result shows bax gene increases in MR.K562,suggests it tend to apoptosis.To analysis the cybrid cells differences in proteomics,we pick up the cells after fusion the 12h,24h,36h,48h,72h five points in time for analysis.Currently have isolated about 40 differences protein points,10 of them were identified by the mass spectrum.