摘要
目的探讨新发现的跨膜丝氨酸蛋白酶4(TMPRSS4)亚型对转染结肠癌(CC)细胞生物学特性的影响。方法采用RT-PCR和Western blot法鉴定转染到人CC细胞(DLD-1)中的命名为T4-1A和T4-1B的TMPRSS4亚型。采用划痕实验和侵袭实验方法阐明稳定型转染细胞的迁移和侵袭能力。结果成功构建T4-1A和T4-1B重组载体,并获得T4-1A和T4-1B转染细胞株,且其T4-1A和T4-1B高表达于稳定型DLD-1。T4-1A与转染真核细胞表达空白载体(pc DNA6)相比,显著促进转染T4-1A的DLD-1迁移和侵袭,差异有统计学意义(P <0. 05),但T4-1B与pc DNA6相比差异无统计学意义。T4-1A和TMPRSS4生物学特性非常相似。结论 T4-1A和T4-1B是新的TMPRSS4亚型,包含激活功能区的T4-1A可促使CC细胞的迁移和侵袭能力,为进一步研究TMPRSS4的分子生物学特性提供理论基础。
Objective To explore a newly discovered transmembrane protease serine 4( TMPRSS4) isoforms and its molecular charcteristics in transfected colon cancer cells. Methods The named T4-1 A and T4-1 B of the TMPRSS4-isoforms were authenticated by the RT-PCR and Western blot,and then transfected to human colon cancer cells( DLD-1). Those stable transfected cells of migration and invasion were illustrated using wound healing assays and matrigel invasion assays. Results Successfully constructed T4-1 A and T4-1 B recombinant vectors,and obtained T4-1 A and T4-1 B transfected cell lines,and their T4-1 A and T4-1 B were highly expressed in stable DLD-1.Compared to cells transfected with empty vector of pcDNA6,the transfected DLD-1 with T4-1 A enhanced migration and invasion were statistical significance( P < 0. 05). However,compared to pcDNA6 no significant1 difference was found for T4-1 B. Moreover,the biological characteristics of T4-1 A and TMPRSS4 were very similar. Conclusion The T4-1 A and T4-1 B is newly TMPRSS4-isoforms,and protease domain included to a T4-1 A has further facilitated the migration and invasion of colon cancer cells,and further studies provide a theoretic base in the molecular biomedical characteristics of TMPRSS4.
作者
赵雪峰
许广大
Zhao Xuefeng;Xu Guangda(Dept of General Surgery,The Affiliated Xinhua Hospital of Dalian University,Dalian 116021)
出处
《安徽医科大学学报》
CAS
北大核心
2019年第2期187-191,共5页
Acta Universitatis Medicinalis Anhui
基金
辽宁省自然科学基金(编号:201602037)
大连大学博士启动基金(编号:20151QL019)
