The Study Of The Anti-gastric Cancer Mechanism Of IL-24 In Vitro And Vivo

Objective:With the eukaryotic expression vector pcDNA3.0-hIL-24 and the prokaryotic expression vector pET-21a(+)-hIL-24, we make them transfect CHO cells and BL-21 respectively and expressed high performance in them. Then assess the bioactive function of the recombinant human IL-24 (rhIL-24) in vitro and in vivo.Methods:Contrive a prime of human IL-24 inserted two restriction enzyme cutting sites KpnⅠand XbaⅠ. Then construct the eukaryotic express vector by PCR using pcDNA3.0 as a template and transfect the CHO cells. Full-length hIL-24 was inserted with BamHⅠand XhoⅠrestriction sites and amplified by PCR using pBV220 as template to construct the prokaryotic expression vector and make it transfect BL-21. Then make the CHO cell and the BL-21 to express rhIL-24 high performance and stably. We detect the expression of rhIL-24 in CHO cells and in BL-21 by SDS-PAGE and western-blot. The apoptosis-inducing activities of rhIL-24 was tested by MTT assay, Hoechst & FCM assay, and the expression of IL-6,TNF-αand IFN-γfrom PBMC induced by rhIL-24 was tested by ELISA. We prove the anti-angiogenesis effect of rhIL-24 by using chick embryo chorioallantoic membrane vascellum model. We observed the inhibitory effects of rhIL-24 and the eukaryotic express vector pcDNA3.0-hIL-24 on model of gastric cancer in nude mice finally. We inject the rhIL-24,pcDNA3.0-hIL-24,CDDP and PBS to the tumor mass every three days. Then observe the changes of tumor growth. The immunohistochemic technique was used to investigated the expression of BCL-2,BAX,VEGF and CD34 in tumor tissues.