摘要
目的:探索转化生长因子β1(tansforming growth factor-β1,TGF-β1)对人成熟树突状细胞(mature dendritic cells,mDCs)骨架丝状肌动蛋白(filament actin,F-actin)及其部分骨架结合蛋白表达的影响,为深入理解树突状细胞(dendritic cells,DCs)的生物学行为和提高基于DCs的抗肿瘤免疫治疗的临床效率提供线索。方法:不同浓度的TGF-β1处理mDCs后,用激光共聚焦显微镜和免疫印迹实验分别研究细胞骨架F-actin的结构和部分细胞骨架结合蛋白表达水平的变化。结果:(1)与对照组相比,TGF-β1处理后的mDCs的F-actin出现了明显重排,F-actin的表达量在3 ng/ml组下调(P=0.000),在5 ng/ml组上调(P=0.000)。(2)mDCs表面丝状突起长度和数量的变化,长度在3 ng/ml和5 ng/ml组较对照组细、短(P=0.001,0.000);数量在1、3 ng/ml和5 ng/ml组较对照组少而稀疏(P=0.000);在7 ng/ml组mDCs表面丝状突起长度和数量的变化均无统计学意义(P=0.114);通过回归分析细胞丝状突起长度和数量与F-actin表达量之间存在非线性相关性(R2分别为0.828和0.746,P=0.000)。(3)细胞骨架蛋白结合蛋白的表达,fascin1在所有实验组中均出现了下调(P=0.001、0.000);p-cofilin1与总cofilin1的表达水平比在1 ng/ml和3 ng/ml组均下调(P=0.000);profilin的表达在1、3 ng/ml和5 ng/ml组均上调(P=0.001、0.001、0.013)。结论:TGF-β1以浓度依赖的方式影响mDCs的细胞骨架F-actin结构及其部分结合蛋白的表达,提示在临床上施行基于DCs的抗肿瘤免疫治疗时,需以适当的方式阻断TGF-β1的信号转导通路,这对进一步深入理解DCs的生物学行为和肿瘤的免疫逃逸机制具有重要意义。
Objective：To explore the effects of transforming growth factor-β1（TGF-β1）on the cytoskeleton filament actin（F-actin） and expression of some binding proteins in mature dendritic cells（mDCs）in order to further understand the biological behaviors and find the clue of improving the clinical efficiency of DCs-based therapy against cancer. Methods：mDCs were treated with different concentrations of TGF-β1 and the organization of cytoskeleton F-actin and expression of some cytoskeleton-binding proteins were respectively observed and measured by laser confocal microscopy and Western blot. Results：（1）F-actin organization was reconstructed in mDCs treated with various concentrations of TGF-β1 compared with that in control group. F-actin expression quantity in 3 ng/ml group was significantly down-regulated（P=0.000）and expression quantity of F-actin in 5 ng/ml group was significantly upregulated（P=0.000）.（2）Length of cell membrane protrusions in 3 ng/ml and 5 ng/ml group was thinner and shorter than that in control group（P=0.001,0.000）. Number of cell membrane protrusions in 1,3,5 ng/ml groups was sparser than that in control group（P=0.000）. Non-liner correlation between expression of Factin and characteristics of cell membrane protrusion（length and number） was observed（R2=0.828,0.746 respectively,P= 0.000）.（3）Expression of cytoskeleton-binding proteins and expression quantity of F-actin in all groups was significantlydown-regulated（P=0.001,0.000）. Phosphorylation of cofilin1 in 1 ng/ml and 3 ng/ml group was significantly down-regulated（P=0.000,0.000）. Expression quantity of profilin in 1,3,5 ng/ml group was significantly up-regulated（P=0.001,0.001,0.013）. Conclusions：Organization of cytoskeleton F-actin and some of its binding proteins in mDCs are affected by TGF-β1 in a concentration-dependent manner,indicating that the signal pathway of TGF-β1 should be blocked in appropriate way before performing DCs-based immunotherapy against cancer. It’s of gre
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2014年第5期646-650,共5页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(编号:11162003
31260227
31170886)
教育部科学技术研究重点资助项目(编号:210196)
贵州省优秀青年科技人才支持计划资助项目(编号:2011-24)
贵州省社会发展科技攻关资助项目(编号:黔科合SY字[2011]3065)
贵州省省长专项基金资助项目(编号:黔省专合字[2009]-79)
贵州省科学技术基金资助项目(编号:黔科合J字2008-2274)
贵阳市科学技术资助项目(编号:2010-筑科农合同字第1-社-12)
