Synergistic effect of rhLIF and IL-24 gene on apoptosis of HL-60 cells / 中国免疫学杂志

ABSTRACT

Objective:To construct the cells which can express LIF protein steadily and study the effect of rhLIF and IL-24 on HL-60 cells.Methods:ECV-304 cells were infected by eukaryotic expression plasmid pcDNA3-LIF,then the positive cells were selected by G418.The positive cells were collected and their culture supernatant was stored for further use.Expression of LIF mRNA was detected by RT-PCR.The pAdeasy-1-pAdTrack-CMV-IL-24 was extracted from DH5a.The recombined adenovirus vector was lineared with PacI and transfected into 293 cells.The IL-24 recombined adenovirus(Ad-IL-24) was obtained and used to infect HL-60 cells.At the same time,the culture supernatant containing rhLIF was added into the HL-60 cells which was identified as positive expression of IL-24 by RT-PCR.Synergistic effect of the cytokines on HL-60 cells was tested by LSCM, FCM and immunohistochemistry stain assay.Results:The cells expressing LIF protein steadily were constructed successfully, the high titer of the recombined adenovirus(Ad-IL-24) was obtained. Expression of IL-24 in infected HL-60 cells was identified by RT-PCR.The apoptosis of HL-60 cells induced by rhLIF and IL-24 was proved by LSCM ,FCM and immunohistochemistry stain assay. They had synergistic effect.Conclusion:rhLIF and IL-24 can inhibit growth of HL-60 cells and induce apoptosis of the cells.They have synergistic effect.