| 稳定表达Trop-2基因的NIH3T3细胞系的构建和特性分析 |
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| 引用本文： | 刘 玉,唐小军,曹 清,丁贵鹏,张慧林,王 欢,郑 峰,徐凛锋,林 红.稳定表达Trop-2基因的NIH3T3细胞系的构建和特性分析[J].南京医科大学学报,2013(7):867-872. |
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| 作者姓名： | 刘 玉 唐小军 曹 清 丁贵鹏 张慧林 王 欢 郑 峰 徐凛锋 林 红 |
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| 作者单位： | 南京军区军事医学研究所,江苏 南京 210002;南京医科大学卫生部抗体技术重点实验室 ,江苏 南京 210029;南京医科大学卫生部抗体技术重点实验室 ,江苏 南京 210029;南京医科大学卫生部抗体技术重点实验室 ,江苏 南京 210029;南京医科大学卫生部抗体技术重点实验室 ,江苏 南京 210029;南京医科大学卫生部抗体技术重点实验室 ,江苏 南京 210029;南京军区军事医学研究所,江苏 南京 210002;南京医科大学卫生部抗体技术重点实验室 ,江苏 南京 210029;江苏省血液中心,江苏 南京 210006 |
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| 基金项目： | 国家自然科学基金资助(81101704);南京市科技局(201201090) |
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| 摘 要： | 目的:建立稳定表达人滋养层细胞表面抗原(Trop-2)NIH3T3细胞,分析过表达Trop-2对NIH3T3细胞的生长?增殖和侵袭特性的影响?方法:将Trop-2基因克隆到真核表达载体pcDNA3.1,转染NIH3T3细胞,通过G418筛选及RT-PCR鉴定获得稳定表达Trop-2的NIH3T3细胞(NIH3T3-Trop-2)?用MTS法检测NIH3T3-Trop-2细胞的增殖能力,软琼脂集落形成实验检测NIH3T3-Trop-2细胞的克隆形成能力,明胶酶谱法检测NIH3T3-Trop-2细胞的基质金属蛋白酶(MMP)-2和MMP-9的分泌及细胞划痕实验检测NIH3T3-Trop-2细胞的迁移能力?结果:稳定表达Trop-2的NIH3T3细胞在生长增殖?克隆形成及侵袭能力均较NIH3T3细胞强,细胞培养上清中的MMP-2和MMP-9增多?结论:Trop-2对细胞的增殖与迁移能力具有明显的促进作用?
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| 关 键 词： | Trop-2 NIH3T3 真核表达 增殖与迁移 |
| 收稿时间： | 2013/2/23 0:00:00 |
Construction and characterization of Trop-2 stable expression in NIH3T3 cells |
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| Liu Yu,Tang Xiaojun,Cao Qing,Ding Guipeng,Zhang Huilin,Wang Huan,Zheng Feng,Xu Linfeng and Lin Hong.Construction and characterization of Trop-2 stable expression in NIH3T3 cells[J].Acta Universitatis Medicinalis Nanjing,2013(7):867-872. |
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| Authors: | Liu Yu Tang Xiaojun Cao Qing Ding Guipeng Zhang Huilin Wang Huan Zheng Feng Xu Linfeng and Lin Hong |
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| Institution: | Huadong Medical Institute of Biotechniques,Nanjing 210002;Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210029;Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210029;Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210029;Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210029;Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210029;Huadong Medical Institute of Biotechniques,Nanjing 210002;Key Laboratory of Antibody Technique of Ministry of Health,NJMU,Nanjing 210029;Jiangsu Blood Center,Nanjing 210006,China |
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| Abstract: | Objective:To establish an NIH3T3 cell line to stable express human Trop-2 gene,and analysis the affection of Trop-2 on proliferation,migration and aggressiveness of NIH3T3-Trop-2 cell. Methods:The human Trop-2 gene was cloned into eukaryotic expression system pcDNA3.1 and transfected into NIH3T3 cells. The Trop-2 stable expression cells were selected by G418 and confirmed by RT-PCR. The cell proliferation,migration and aggressiveness were detected by MTS and wound healing assay,MMP-2/MMP-9 was analyzed. Results:The proliferation of NIN3T3-Trop-2 was higher than NIN3T3 cell. The wound healing assay results showed and that Trop-2 improved the cell migration,increased the number of foci generated(P < 0.05) and increased MMP-9 expression(P < 0.05) when compared to the NIH3T3 control group. Conclusion:These show that human Trop-2 is sufficient to improve the cell proliferation and induce the transformation of NIH3T3 cells. |
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| Keywords: | Trop-2 NIH3T3 eukaryotic expression proliferation and migration |
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