CIITA反义RNA抑制MHCII类分子表达

Suppression of MHC Class II Molecules by Antisense RNA of MHC Class II Transactivator

为探索利用CIITA (MHCclassIItransactivator)基因反义RNA抑制MHCII类分子表达的可能性 ,为CIITA的应用研究奠定基础。利用RT PCR扩增能与CIITAcDNA第 95至 5 0 0bp互补的片段 ,并构建成pcDNA3/CIITAa 重组质粒 ,脂质体转染法将其转入人HeLa/Raji细胞和猪PIEC/L2 3细胞 ,流式细胞术动态检测反义RNA对人 /猪MHCII类分子的抑制率和抑制程度。结果显示HeLa、Raji、PIEC和L2 3四种细胞MHCII类分子受抑制率分别为 4 6 7% ( 7/ 15 ) ,4 0 % ( 6 / 15 ) ,4 6 7% ( 7/ 15 )和33 3% ( 5 / 15 )。典型受抑克隆细胞MHCII类分子受抑程度高达 85 % ,反义RNA对MHCII类分子的抑制时间持续 35～ 5 0d。CIITA反义RNA能有效抑制人 /猪MHCII类分子的表达 。

To explore the possibility of suppression of MHC class II molecules by antisense RNA of MHC class II transactivator (CIITA),the fragment in 405 bp length complementary to CIITA cDNA was amplified,and the plasmid pcDNA3/CIITA was constructed Then the plasmid was transfected into HeLa cells,Raji cells, PIEC and L23 cells by using lipofectamine Flow cytometry analysis was used to detect the degree of suppression of MHC class II molecules It was found that the expression of MHC class II molecules in 7 of 15 selected HeLa cells,6 of 15 selected Raji cells,7 and 5 of 15 selected PIEC and L23 cells were suppressed,of which the percentages of suppression might be up to 85%,and the suppression could maintain for 35～50 days It concludes that antisense RNA of CIITA can suppress the expressions of MHC class II molecules,and this may be useful in the researches of autoimmunity and transplantation immunology