The Expression of M-CSF in NIH3T3 Cell Nucleus and its Effect on Cell Movement

Objectives To construct macrophage colony-stimulating factor(M-CSF)-expressing vector (pCMV/M-CSF) and explore the effect of nuclear M-CSF on the NIH3T3 cells movement. Methods Function part of M-CSF cDNA was amplified by PCR and inserted to nucleus-localization expression vector pCMV/myc/nuc to construct pCMV/M-CSF. Then the pCMV/M-CSF was transfected into NIH3T3 cells and the cells clones were selected with G418. The expression and localization of M-CSF in NIH3T3 cells were verified by immunocytochemistry and Western blot. At last,the effect of nuclear M-CSF on cell movement was analyzed by cell scratch assay. Results The size of inserted fragments in the recombinant vectors pCMV/M-CSF is 1400bp, corresponding to that of M-CSF. After transfecting pCMV/M-CSF into NIN3T3 cells by liposome and screening using G418, pCMV/M-CSF-transfected NIN3T3 cells can stably express M-CSF protein in nucleus. The results from cell scratch assay show that pCMV/M-CSF-transfected NIN3T3 cells have stronger movement ability. Conclusions The nuclear localization vector pCMV/M-CSF was successfully constructed, M-CSF in nucleus promoted the NIH3T3 cells movement.