Construction of murine mammary cancer cell line expressing IL-23 gene and its antitumor effects

Objective To construct the murine mammary cancer cell line IL-23/MA-891 ex- pressing IL-23 protein,and investigate the inhibitory effects of/L-23 on the growth of tumor cells in vitro and in vivo.Methods The IL-23 gene was packed with two packing cell lines(ecotropicψ2 and ampho- tropic PA317)by a retrovirus vector(LXSN),and the positive cell clones were screened by G418.After transfection to MA-891 cells with the culture supernatant of IL-23/PA317 cells and selection with G418, the IL-23/MA-891 cells expressing IL-23 were obtained.The expression of IL-23 mRNA and protein was detected by RT-PCR and laser scanning confocal microscope(LSCM),respectively.The ability of IL-23/ MA-891 culture supernatant inducing splenocytes to secrete IFN-γwas detected with ELISA.The prolifera- tion of MA-891,LXSN/MA-891 and IL-23/MA-891 cells was detected by MTT colorimetry and flow cy- tometry in vitro.Mice were subcutaneously inoculated with MA-891 cells which were transfected with IL-23 or empty vector(LXSN),and the tumor size in mice was measured.The inhibitory effect of IL-23 gene transfection on mouse solid tumor was observed.Results IL-23/MA-891 cells stable expressed IL-23 mRNA and protein.The culture supernatant of IL-23/MA-891 cells induced secretion of IFN-γby mouse splenocytes.The proliferation rate of IL-23/MA-891 cells was slightly decresed(P0.05),and the per- centage of mice with detectable tumor showed no significant difference after subcutaneous inoculation of MA-891 cells transfected with IL-23.Growth rate of tumor in mice model was also inhibited in IL-23 gene therapy group as compared with the control group(P0.05).Conclusion IL-23/MA-891 cell line ex- pressing IL-23 was successfully constructed.In in vivo,IL-23 can remarkably inhibit the growth of solid tumor in mice.