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| 抗PNAS-2单链抗体对白血病U937细胞凋亡的影响 | |
| 引用本文： | 肖予,陈芳源,王海嵘,吴英理,钟济华,钟华. 抗PNAS-2单链抗体对白血病U937细胞凋亡的影响[J]. 上海交通大学学报(医学版), 2011, 0(4): 387-391 |
| 作者姓名： | 肖予 陈芳源 王海嵘 吴英理 钟济华 钟华 |
| 作者单位： | 上海交通大学医学院附属仁济医院血液科;上海交通大学医学院细胞分化与凋亡教育部重点实验室; |
| 基金项目： | 仁济医院-交通大学基础医学院合作研究基金(ZD0704); 国家自然科学基金(81000212)~~ |
| 摘 要： | 目的观察抗PNAS-2单链抗体与PNAS-2抗原特异性结合的能力,并在此基础上探究其对急性单核细胞白血病U937细胞凋亡的影响。方法运用反转录病毒转染法将抗PNAS-2单链抗体表达载体pMIG-PNAS-2ScFv转入U937细胞(pMIG-PNAS-2ScFv组),同时设转染pMIG空载体(NC组)及未转染载体的U937细胞(U937组)为对照。流式细胞仪检测转染效率;激光共聚焦显微镜观察单链抗体表达情况及与靶抗原特异性结合的能力;流式细胞仪和激光共聚焦显微镜检测细胞凋亡;Western blotting观察PNAS-2表达及促凋亡因子caspase-3激活情况。结果成功将抗PNAS-2单链抗体编码序列转入U937细胞,所表达的单链抗体能与PNAS-2抗原特异性结合。与NC组和U937组相比,pMIG-PNAS-2ScFv组细胞凋亡率明显上升(P<0.05),活化型caspase-3表达增加,但PNAS-2含量并无变化。结论抗PNAS-2单链抗体能有效结合U937细胞内的PNAS-2抗原并有促进U937细胞凋亡的作用,其促凋亡机制可能与PNAS-2功能位点被封闭后功能失活而无法发挥抑凋亡作用有关。 |
| 关 键 词： | PNAS-2 单链抗体 U937细胞 凋亡 |
Effects of anti-PNAS-2 single chain antibody on apoptosis of human leukemic U937 cells |
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| XIAO Zhen-yu,CHEN Fang-yuan,WANG Hai-rong,WU Ying-li,ZHONG Ji-hua,ZHONG Hua. Effects of anti-PNAS-2 single chain antibody on apoptosis of human leukemic U937 cells[J]. Journal of Shanghai Jiaotong University:Medical Science, 2011, 0(4): 387-391 | |
| Authors: | XIAO Zhen-yu CHEN Fang-yuan WANG Hai-rong WU Ying-li ZHONG Ji-hua ZHONG Hua |
| Affiliation: | XIAO Zhen-yu1,CHEN Fang-yuan1,WANG Hai-rong1,WU Ying-li2,ZHONG Ji-hua1,ZHONG Hua1(1.Department of Hematology,Renji Hospital,Shanghai Jiaotong University School of Medicine,Shanghai200127,China,2.Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education,Shanghai200025,China) |
| Abstract: | Objective To observe specific binding ability of anti-PNAS-2 single chain antibody with PNAS-2 antigen and its effects on apoptosis of human leukemic U937 cells. Methods The pMIG-PNAS-2ScFv,anti-PNAS-2 single chain antibody expression vector,was transfected into U937 cell line by retroviral transfection method.Untransfected U937 cells and pMIG transfected with empty vector were used as controls.Transfection rate was measured by flow cytometry.Single chain antibody expression and ScFv-antigen specific bindin... |
| Keywords: | PNAS-2 single chain antibody U937 cell apoptosis |
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