ABSTRACT
Objective :To construct hepatocarcinoma specific IL-1? anti-sense RNA expression vector and to explore its effect on the growth of implanted hepatocarcinoma H22 cells in mice and the possible mechanism. Methods:Murine IL-1? anti-sense RNA expression vectors pafpIRES2-antiIL-1?1 and pafpIRES2-antiIL-1?2 under the regulation of minimal alpha-feto protein (AFP) promoter and CMV enhancer were constructed,and further verified by PCR,restriction endonuclease analysis and DNA sequencing. H22 cells transfected with pafpIRES2-antiIL-1? 1 or pafpIRES2-antiIL-1? 2 were divided into 3 groups:H22/mock,H22/antiIL-1?1 and H22/antiIL-1?2 group. Expression of IL-1? was detected by RT-PCR. Transfected H22 cells were subcutaneously injected into mice to establish tumor implanted mouse model. Tumor volume was measured; the cytotocixity of spleen NK against H22 cells was detected by MTT. Results:Hepatocarcinoma specific IL-1? anti-sense RNA expression vectors pafpIRES2-antiIL-1?1 and pafpIRES2-antiIL-1?2 were successfully constructed and were verified by PCR,restriction endonuclease analysis and DNA sequencing. IL-1? expression in H22 cells was down-regulated after transfected with IL-1? anti-sense RNA expression vectors,especially with the pafpIRES2-antiIL-1?2 vector. Hepatocarcinoma cells implanted mouse model was successfully established. Tumor volume and growth of tumor in H22/antiIL-1?2 mice was obviously smaller than that in H22/mock mice,and the cytotocixity of spleen NK against H22 cells in H22/antiIL-1?1 and H22/antiIL-1?2 mice was also greatly enhanced. Conclusion:Hepatocarcinoma specific IL-1? anti-sense RNA expression vector pafpIRES2-antiIL-1? was successfully constructed. It effectively inhibits the growth of implanted hepatocarcinoma in mice probably through specifically blocking expression of IL-1? and increasing cytotocixity of spleen NK.