| 抗MRP3单链抗体-sTRAIL融合蛋白诱导U251多形性胶质母细胞瘤凋亡 |
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| 引用本文： | 倪长伟,赵辰阳,郎悦,苏静,乐媛,吕翠婷,王梁华,尹剑.抗MRP3单链抗体-sTRAIL融合蛋白诱导U251多形性胶质母细胞瘤凋亡[J].中国生物化学与分子生物学报,2012,0(8):761-767. |
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| 作者姓名： | 倪长伟 赵辰阳 郎悦 苏静 乐媛 吕翠婷 王梁华 尹剑 |
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| 作者单位： | 大连医科大学附属第二医院神经内科;大连医科大学附属第二医院神经外科;第二军医大学生物化学与分子生物学教研室 |
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| 基金项目： | 国家新药创制科技重大专项课题(No.2009ZX09102-234,No.2009ZX09103-689);国家自然科学基金项目(No.30571906)~~ |
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| 摘 要： | 采用重组PCR技术获得抗多药耐药相关蛋白3(multidrug resistance protein 3, MRP3)的单链抗体(scFv)与人源可溶性肿瘤坏死因子相关凋亡诱导配体(soluble TNF-related apoptosis inducing ligand, sTRAIL)的融合蛋白质的基因编码序列, 利用原核表达载体pMAL-c2,构建含麦芽糖结合蛋白(maltose binding protein, MBP)标签肽的antiMRP3(scFv)-sTRAIL融合蛋白, 经亲和层析柱纯化. 获得纯化的antiMRP3(scFv)-sTRAIL融合蛋白，用MRP3阳性U251多形性胶质母细胞瘤做增殖抑制实验、细胞凋亡诱导实验，结果均显示具有明显的活性, 而MBP无明显作用. 上述结果表明，成功表达了antiMRP3(scFv)-sTRAIL融合蛋白, 该融合蛋白具有诱导U251多形性胶质母细胞瘤细胞凋亡的活性, 为开发靶向性抗肿瘤药物奠定了基础.
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| 关 键 词： | 单链抗体 多药耐药相关蛋白3 肿瘤坏死因子相关凋亡诱导配体 多形性胶质母细胞瘤 |
| 收稿时间： | 2012-02-27 |
Induction of Apoptosis in U251 Polymorphic Glioblastoma Cells by Recombinant AntiMRP3(scFv)-sTRAIL Fusion Protein |
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| NI Chang-Wei,ZHAO Chen-Yang,LANG Yue,SU Jing,LE Yuan,LV Cui-Ting,WANG Liang-Hua,YIN Jian.Induction of Apoptosis in U251 Polymorphic Glioblastoma Cells by Recombinant AntiMRP3(scFv)-sTRAIL Fusion Protein[J].Chinese Journal of Biochemistry and Molecular Biology,2012,0(8):761-767. |
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| Authors: | NI Chang-Wei ZHAO Chen-Yang LANG Yue SU Jing LE Yuan LV Cui-Ting WANG Liang-Hua YIN Jian |
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| Institution: | 1)Second Affiliated Hospital of Dalian Medical University,Department of Neurology,Dalian 116027,Liaoning,China; 2)Second Affiliated Hospital of Dalian Medical University,Department of Neurosurgery,Dalian 116027,Liaoning,China; 3)Department of Biochemistry and Molecular Biology,Second Military Medical University,Shanghai 200433,China) |
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| Abstract: | Both gene fragments of single-chain antibody fragment(scFv) against multidrug resistance protein 3(MRP3) and human soluble TNF-related apoptosis inducing ligand(sTRAIL) were amplified by PCR,and then inserted into a pMAL-c2 prokaryotic expression vector.The fusion protein of antiMRP3(scFv)-sTRAIL was expressed by transforming E.coli BL21 with IPTG induction,then purified with amylose resin chromatography by its maltose binding protein(MBP) tag.The obtained antiMRP3(scFv)-sTRAIL fusion protein substantially inhibited the proliferation of MRP3-positive U251 glioblastoma cells,and induced apoptosis significantly.The results indicated that recombinant antiMRP3(scFv)-sTRAIL fusion protein obtained from E.coli expression exhibited potent apoptosis-inducing effect in U251 cells. |
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| Keywords: | single-chain antibody fragment multidrug resistance protein 3 TNF-related apoptosis inducing ligand polymorphic glioblastoma multiforme |
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