| 邓侃,陈廷涛,彭颖征.靶向重组Aif-T18融合蛋白对TEM1阳性肿瘤细胞的杀伤作用的研究[J].井冈山大学自然版,2021,42(1):43-48 |
| 靶向重组Aif-T18融合蛋白对TEM1阳性肿瘤细胞的杀伤作用的研究 |
| EVALUATION OF KILLING EFFECT OF AIF-T18 FUSION PROTEIN ON TEM1 POSITIVE CANCER CELLS |
| 投稿时间：2020-10-22 修订日期：2020-12-02 |
| DOI：10.3969/j.issn.1674-8085.2021.01.008 |
| 中文关键词: TEM1 单链抗体 Aif |
| 英文关键词: TEM1 single chain antibody Aif |
| 基金项目:国家自然科学基金项目（31360377）；江西省教育厅青年科学基金项目（GJJ14197，GJJ14218） |
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| 中文摘要: |
| 验证Aif-T18融合蛋白对肿瘤内皮细胞标志分子1（TEM1）阳性肿瘤细胞的杀伤作用。采用基因工程技术方法构建重组质粒pIRES-TEM1-EGFP，转染TEM1阴性表达细胞MS1，经G418筛选得到TEM1阳性细胞株（MS1-TEM1）；通过构建重组质粒pET302-Aif和pET302-Aif-T18，转化至表达宿主菌BL21中，经异丙基-β-D-硫代吡喃半乳糖苷（IPTG）诱导菌体得到所需的目的蛋白，采用流式细胞仪和MTT技术检测融合蛋白对MS1和MS1-TEM1细胞的亲和力和特异性杀伤的效果。本研究中，主要利用单链抗体scFvT18选择性携带凋亡诱导因子AIF，靶向抑杀TEM1阳性细胞，从而为Aif-T18融合蛋白在肿瘤治疗中的开发应用提供实验依据。 |
| 英文摘要: |
| To verify the killing effect of Aif-T18 fusion protein on tumor endothelial cell marker molecule 1 (TEM1) positive tumor cells, genetic engineering technology was used to construct the recombinant plasmid pIRES-TEM1-EGFP, transfected with TEM1-negative expression cell MS1, and screened by G418 to obtain TEM1-positive cell lines (MS1-TEM1). The recombinant plasmids pET302-Aif and pET302-Aif-T18, transformed into the expression host strain BL21, and the target protein was induced by bacterial cells induced by isopropyl-β-D-thiogalactopyranoside (IPTG). The affinity and specific killing effect of the fusion protein on MS1 and MS1-TEM1 cells were detected by flow cytometry and MTT. In this study, the single-chain antibody scFvT18 was used to carry the apoptosis-inducing factor AIF selectively and target and suppress TEM1-positive cells, thereby providing an experimental basis for the development and application of Aif-T18 fusion protein in tumor therapy. |
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