Smad3小干扰RNA在转化生长因子β_1作用下对树突状细胞成熟及功能的影响  

Effects of Smad3 SiRNA on Maturity and Function of Dentritic Cells with Transforming Growth Factor-β_1

作　　者：马楠[1] 祁馨卉[1] 崔慧霞[1] 李妍[1] 刘云鹏[1] 姜又红[1] 

机构地区：[1]中国医科大学附属第一医院肿瘤研究所二室,沈阳110001

出　　处：《中国医科大学学报》2012年第10期926-929,共4页Journal of China Medical University

基　　金：辽宁省科学技术计划项目(2011415052-3)

摘　　要：目的研究通过RNA干扰技术阻断外周单核细胞来源的树突细胞(DC)内的Smad3,并在转化生长因子β(1TGF-β1)作用下对DC的成熟和免疫抑制作用的影响。方法设计针对Smad3特异性小干扰RNA(siRNA),利用RNAiMAX转入DC,并用RT-PCR检测转染效果。实验分Control-DC组、TGF-β1-Control-DC组、TGF-β1-Negative siRNA-Smad3 DC组和TGF-β1-siRNA-Smad3 DC组。流式检测各组DC成熟表型的变化,ELISA检测各组DC上清白细胞介素12(IL-12)的水平,CCK-8法检测DC刺激同种异体T细胞增殖的能力。结果 TGF-β1-siRNA-Smad3 DC组表面成熟标志CD80、CD83、CD86、HLA-DR及IL-12分泌水平明显高于TGF-β1-Control-DC组和TGF-β1-Negative siRNA-Smad3 DC组(P<0.05),且TGF-β1-Negative siRNA-Smad3 DC组刺激同种异体T细胞增殖的能力显著增强。结论通过阻断DC的Smad3表达,可以逆转TGF-β1对DC成熟和免疫功能的抑制。Objective To investigate the effects of small interference RNA（siRNA） targeting Smad3 on the maturity and function of dentrictic cell（DC） with transforming growth factor-β1（TGF-β1）.Methods The siRNA sequences targeting Smad3 gene were designed,and transfected to DC by RNAiMAX.The transfection efficiency was detected by RT-PCR.The experimental classification was performed as followed：group of Control-DC,group of TGF-β1-Control-DC,group of TGF-β1-Negative siRNA-Smad3 DC and group of TGF-β1-siRNA-Smad3 DC.The maturity of DC was investigated by FCM.The level of IL-12 was determined by ELISA and the proliferation of allogenic T cell stimulated with DC was detected by CCK-8 assays.Results Expression of CD80,CD83,CD86 and HLA-DR and level of IL-12 in group of TGF-β1-siRNA-Smad3 DC was higher than both TGF-β1-Control-DC group and TGF-β1-Negative siRNA-Smad3 DC group（P 0.05）.The group of TGF-β1-siRNA-Smad3 DC had higher stimulation on the allogenic T cell proliferation.Conclusion Block Smad3 expression in DC by siRNA can reverse the TGF-β1 inhibition on the maturity and immune function of DC.

关 键 词：树突状细胞 转化生长因子β1 SMAD3 肿瘤疫苗 

分 类 号：R730.51[医药卫生—肿瘤]