SDF-1γ/rhGM-CSF融合蛋白的制备及其趋化作用  

Preparation of SDF-1γ/rhGM-CSF fusion protein and its chemotactic effect

作　　者：居小萍[1] 徐新颜[2] 张晓青[1] 刘永明[1] 于春山[1] 曹洋森[1] 卢明智[1] 陈樱[1] 

机构地区：[1]第二军医大学长海医院放射治疗科,上海200433 [2]中国科学院上海生命科学研究院植物生理生态研究所,上海200032

出　　处：《中国肿瘤生物治疗杂志》2011年第4期419-423,共5页Chinese Journal of Cancer Biotherapy

基　　金：上海市自然科学基金资助项目(No.06ZR14114);军队医学研究"十一五"计划课题资助项目(No.10MA004)~~

摘　　要：目的:利用基因工程技术制备SDF-1与hGM-CSF的融合蛋白(SDF-1γ/rhGM-CSF),研究该融合蛋白对肿瘤患者造血和免疫功能的增强作用。方法:构建表达SDF-1γ/rhGM-CSF融合蛋白的pPIC9k-SDF1-rhGM-CSF1质粒,转染酵母菌,诱导SDF-1γ/rhGM-CSF融合蛋白的表达,Western blotting鉴定SDF-1γ/rhGM-CSF融合蛋白的表达。集落形成实验观察SDF-1γ/rhGM-CSF对骨髓细胞集落形成的影响,趋化实验检测其对未成熟树突状细胞(dendritic cell,DC)的趋化作用。结果:成功构建pPIC9k-SDF1-rhGM-CSF1质粒,高表达SDF-1γ/rhGM-CSF融合蛋白,分子量约为25 000,并可被GM-CSF特异性抗体所识别。SDF-1γ/rhGM-CSF融合蛋白能显著刺激骨髓细胞的集落形成,其效果强于GM-CSF(P<0.05)。与SDF-1相比,SDF-1γ/rhGM-CSF融合蛋白可更有效地趋化未成熟DC(P<0.05)。结论:SDF-1γ/rhGM-CSF融合蛋白可有效促进骨髓细胞的集落形成,趋化未成熟DC,有促进肿瘤化疗患者造血和免疫功能恢复的潜在临床应用前景。Objective：To prepare the fusion protein of SDF-1 and rhGM-CSF （SDF-1 γ/rhGM-CSF） by genetic engi- neering technology, and investigate its hematopoietic and immune promotion functions in tumor patients. Methods： The expression vector for SDF-1γ/rhGM-CSF fusion protein, pPIC9k-SDFI-rhGM-CSFI, was constructed and the protein expression was induced by yeast transfection. SDF-1γ/rhGM-CSF fusion protein was further identified by Western blotting analysis. Colony-formation assay and chemoattract assay were used to study the roles of the prepared fusion protein in stim- ulating bone marrow cell colony-formation and in chemoattracting immature dendritic cells. Results： SDF-1 γ/rhGM-CSF fusion gene vector, pPIC9k-SDFI-rhGM-CSF1, was successfully constructed and expressed high level of SDF-1γ/rhGM- CSF fusion gene. The molecular weight of the expressed protein was about 25 000 and was recognized by GM-CSF specific antibody. The fusion protein had a stronger effect in stimulating bone marrow cell colony-formation than GM-CSF （P 〈 0. 05 ） and in chemoattracting immature dendritic cells than SDF-1 （P 〈 0.05）. Conclusion ： SDF-1 γ/rhGM-CSF fusion protein can promote bone marrow cell colony-formation and chemoattraction of immature dendritic cells, which might be used for promoting hematopoiesis and immune function of tumor patients after chemotherapy.

关 键 词：SDF—1 GM—CSF 融合蛋白 肿瘤 造血 免疫 趋化 

分 类 号：Q78[生物学—分子生物学] R392.1[医药卫生—免疫学]