稳定表达Trop-2基因的NIH3T3细胞系的构建和特性分析  

Construction and characterization of Trop-2 stable expression in NIH3T3 cells

作　　者：刘玉[1] 唐小军[2] 曹清[2] 丁贵鹏[2] 张慧林[2] 王欢[2] 郑峰[1] 徐凛锋[2] 林红[3] 

机构地区：[1]南京军区军事医学研究所,江苏南京210002 [2]南京医科大学卫生部抗体技术重点实验室,江苏南京210029 [3]江苏省血液中心,江苏南京210006

出　　处：《南京医科大学学报（自然科学版）》2013年第7期867-872,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助(81101704);南京市科技局(201201090)

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。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.

关 键 词：Trop-2 NIH3T3 真核表达 增殖与迁移 

分 类 号：R392.12[医药卫生—免疫学]