Effects of IL-2R antisense RNA expression plasmids' transfection on the proliferation of mouse splenocytes in vitro / 中国药理学通报

ABSTRACT

AIM: To investigate the effects of tranfection of IL-2R antisense RNA expression plasmids on mouse spleen cells' proliferation in vitro and its possible mechanism. METHODS: Spleen cells were transfected with IL-2R antisense RNA eukaryotic expression plasmids using adhesion-assisted lipofection method, and then the spleen cells were stimulated by mitogen. Cells' proliferation was tested by tetrazolium salt (MTT) method. IL-2R mRNA and protein expression level were measured by slot-blot hybridization assay and flow cytometry method respectively. RESULTS: The proliferation of spleen cells was inhibited obviously after transfecting with recombinant plasmids. The inhibitory rate of pcAnti-mlL-2Rαβ- and pciAnti-mIL-2Rαβ-transfected group was higher than that of pcAnti-mIL-2Rα-and pcAnti-mIL-2Rβ-transfected group; the inhibitory rate of pcAnti-mIL-2Rα-tranfected group was higher than that of pcAnti-mlL-2Rβ-tranfected group. No inhibitory effect on the growth of NIH3T3 cells was observed when they were transfected with recombinant plasmids. IL-2R mRNA and protein expression level were decreased in spleen cells after transfection of recombinant plasmids. CONCLUSION: IL-2R antisense RNA can efficiently inhibit the proliferation of mouse spleen cells in vitro. IL-2Rαβ chimeric antisense RNA showed higher inhibitory rate than a or antisense RNA. IL-2Rα antisense RNA was more effective than β antisense RNA. It can be concluded preliminarily that the inhibitory effect of IL-2R antisense RNA was exclusively on the growth of cells functionally expressing IL-2R. The inhibitory effect on the spleen cells' proliferation was likely due to the blocking of IL-2R expression by antisense RNA.