重组超抗原SEB-scFv融合蛋白体外靶向抑制胃癌细胞的研究  

Inhibitory effect of SEB-scFv fusion protein on gastric cancer ceils in vitro

作　　者：舒晓刚[1] 童强[1] 卢晓明[1] 陶凯雄[2] 王国斌[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院胃肠外科,武汉430022 [2]华中科技大学同济医学院附属协和医院腔镜外科,武汉430022

出　　处：《中华实验外科杂志》2008年第4期434-436,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省卫生厅资助项目（NX200513）

摘　　要：目的观察SEB-scFv融合蛋白对人胃癌细胞的体外抑制作用。方法噻唑蓝（MTT）法观察SEB-scFv融合蛋白对人胃癌细胞的增殖抑制作用；DNA ladder法观察融合蛋白对胃癌细胞凋亡的影响，电子显微镜法观察融合蛋白处理后胃癌细胞的形态变化；流式细胞仪分析胃癌细胞凋亡率。结果胃癌细胞的增殖抑制率随SEB-scFv融合蛋白作用浓度的增加而逐渐增大，且对不表达MG7相关抗原的肿瘤细胞不表现杀伤效应。1．0、10mg／L高浓度融合蛋白组DNA凝胶电泳呈现明显的梯状条带。被融合蛋白活化的免疫效应细胞攻击后的胃癌细胞表现出典型的凋亡特征性变化。融合蛋白0．1、1．0、10mg／L组细胞G0／G1期前有明显的凋亡峰出现，三者凋亡率分别达到51．62％、54．21％和61．55％。而生理盐水组凋亡率仅为1．44％，差异有统计学意义（P〈0．05）。结论SEB-scFv融合蛋白在效应细胞介导下，能对表达MG7相关抗原的胃癌细胞显示出有效的体外毕长抑制作用．且在此讨程中伴随有胃癌细胞的凋亡。Objective To evaluate the antitumor effect of SEB-scFv fusion protein on gastric cancer cell line SGC7901 in vitro and study the apoptosis of SGC7901 induced by SEB-seFv fusion protein. Methods MTT assay was used to observe the inhibitory effects of SEB-scFv fusion protein on proliferation of SGC7901 cells. By using DAN ladder and flow cytometry,the effects of SEB-scFv fusion protein on the apoptosis of SGC7901 cells were analyzed. The morphologic changes of SGC7901 cells treated with SEB-scFv fusion protein were observed. Results Proliferation inhibitory rate of SGC7901 cells was increased with the increased concentration of SEB-scFv fusion protein in vitro. Electric microscope revealed that the cell presented some typical changes of apoptosis,including cell shrinkage and rounding;cytoplasm appears dense, and the organelles appear tightly packed; pyknosis, karyorrhexis, blebs, apoptotic bodies appear etc. FCM indicated that the apoptotic rate of SGC7901 cell significantly evaluated with increasing dose of SEB-scFv fusion protein, and agarose gel electrophoresis appeared marked DNA ladder, Conclusion SEB-scFv fusion protein can target the effeetor cells to the tumor cells ,and it cotdd induce a dose-dependent apoptotic cell death in SGC7901 cells.

关 键 词：胃癌 超抗原 融合蛋白 脱噬作用 

分 类 号：R735.2[医药卫生—肿瘤]