以GFP为标记的肝癌细胞RNA转染树突状细胞的体外效应论文.doc

以GFP为标记的肝癌细胞RNA转染树突状细胞的体外效应论文.doc以GFP为标记的肝癌细胞RNA转染树突状细胞的体外效应论文
张利旺,张红梅,刘文超,潘伯荣,斯晓明,任军
【关键词】树突状细胞;绿色荧光蛋白;RNA;转染
Effect of dendritic cells transfected arker in vitro
【Abstract】 AIM: To investigate the feasibility of GFP as a marker to observe the dendritic cells (DCs) transfected or cells and the feasibility of the transfected DCs serving as a ine for potential immunotherapy. METHODS: Plasmid pGFPC3 the HepG2GFP using Trizol; DCs ononuclear cells (PBMCs), and transfected icroscope, the changes of phenotype etry, and the change of IL12 secretion in the supernatant of DCs icroscope, so did DCs transfected embrane molecules such as CD80 (% to %), HLADR (% to %), CD83 (% to %), CD86 (% to %) atically, IL12 secretion in the supernatant arker to observe the effect of transfection of DCs or cell RNA. DCs transfected or cell RNA may serve as a promising ine of immunotherapy.
【Keya, HCC)是一种常见的恶性肿瘤,适合手术治疗的患者只占一少部分,具有较高的复发率,预后很差,严重威胁人类健康[1]. DCs(dendritic cells, DCs)是体内功能强大的专职性抗原提呈细胞,它能够高效的摄取、处理抗原,并将处理后的多肽呈递给静息型T细胞,引起针对该抗原的特异性免疫反应[2]. 国内外类似的研究中,都没有明确的可以观察肿瘤RNA转染DCs效果的指标,我们应用绿色荧光蛋白质粒载体 pGFPC3稳定转染肝癌细胞HepG2,提取筛选后细胞HepG2GFP总RNA转染DCs,观察转染后DCs绿色荧光蛋白表达及特异性表面标志及功能相关分子表达,检测其上清细胞因子分泌,探讨其作为肿瘤疫苗的可行性.
1材料和方法
;HepG2肝癌细胞株由我科常规传代培养. 流式细胞仪(BD),DG3022A型酶联免疫检测仪(华东电子管厂),荧光显微镜Optiphot XIF(Nicon)等均为我院常备. D80,FITCHLADR,PECD83,PECD86抗体购自BD公司;rhGMCSF,rhIL4购自Peprorotec公司;TNF