Establishment Of MIN6 Cell Line With Stable Overexpression Of C-Myc Through Retrovirus Infection

c-Myc, a oncogene, is required for the development of tissues and organs. c-Myc promotes or inhibist the expression of target genes through specificially or nonspecifically binding the cis-element of genes to regulate the cellular growth and differentiation. It has been reported that c-Myc promots cell proliferation and invasion through gene amplification in chromosome or activation mutation, thus causes the occurrence and development of pancreatic cancer. Overexpression of c-Myc inβcells leads to apoptosis and decrease of P cells amount in human. To explore the mechanism of regulation of c-Myc onβcells, this study constructed retrovirus vector PMXs-neo-c-myc, package the retrovirus with 293GP cells, and infected theβcell line MIN6 cells. MIN6 clones with stable overexpression of c-Myc gene were got through G418 selection and clonal amplification. The further analysis of MIN6 cells with c-Myc overexpression in the changes of cellular activity was performed.The results showed that multiple MIN6 cell colonies with the integration of c-myc gene were obtained through screening by G418 resistance to drugs; RT-PCR results showed that most of these colonial cells stably overexpressed c-Myc, also they were found the overexpression of miR-17-92 gene cluster; through the analysis of growth curve, we found that overexpression of c-Myc can cause accelerated growth of MIN6 cells, cell cycle time is shorter; through cell staining, we found that apoptosis in some colonies with over-expression of c-Myc significantly increased. The mechanism of changes in cell activities after overexpression of c-Myc should be further explored in the future.