Retrovirus mediated IL-23 gene expression in mouse high-metastasis mammary cancer cells MA-891 / 中国免疫学杂志

ABSTRACT

Objective:The mouse mammary cancer cell line IL-23/MA-891 expressing IL-23 protein was set up and the effect of IL-23 on growth and other biological character of the cells was studied.Methods:The IL-23 gene was sequencely transfected into two packing cell lines (ecotropic ?2 and amphotropic PA317) by a retrovirus vector (LXSN),and the positive cellular clones were screened by G418.After infection to MA-891 cells with the culture supernatant of IL-23/PA317 cells and selected with G418,the IL-23/MA-891 cells to express IL-23 mRNA and protein was detected with RT-PCR,ELISA and immunocytochemical stairing,respectively.The expression of H-2Kb(MHCⅠ),I-Ab(MHC Ⅱ),CD80,CD86 and FAS was examined with flow cytometry.The ability to secrete IFN-? by mouse splenocytes stimulated with the culture supernatant of IL-23/MA-891 cells was detected with ELISA.The proliferation of MA-891,LXSN/MA-891 and IL-23/MA-891 cells was detected by MTT colorimetry and flow cytometry in vitro.Results:IL-23/MA-891 cells express IL-23 stably in mRNA and protein level.The expressing of H-2Kb(MHCⅠ),I-Ab( MHCⅡ),CD80,CD86 and FAS protein by flow cytometry were showing no difference in three kinds of cells.The proliferation rate of IL-23/MA-891 cells were showing little decresed,but statistically showing no difference with parental cells.The culture supernatant of IL-23/MA-891 cells induced secretion of IFN-? by mouse splenocytes were increased compared to parental cells.Conclusion:The cellular clone IL-23/MA-891 with high level expression of IL-23 was produced.