摘要
目的构建TM4SF1的真核表达载体并研究其对结直肠癌细胞株HCT116、DLD1迁移及侵袭功能的生物学特性影响。方法根据GenBank:BC034145.1中的TM4SF1的序列设计两条引物,用逆转录的方法获得开放阅读框(ORF区),并将其连接到质粒上,经鉴定后瞬时转染结直肠癌细胞株HCT116及DLD1,应用Western blot及细胞免疫化学方法检测其转染效果及表达情况,采用划痕及transwell实验观察其对细胞迁移及侵袭能力的影响。结果成功构建PEZ-M29/TM4SF1真核表达载体,Western blot及细胞免疫化学实验结果显示TM4SF1表达上调可以促进结直肠癌细胞株的迁移及侵袭能力。结论成功构建TM4SF1真核表达载体,TM4SF1的上调表达可提高结直肠癌细胞株HCT116、DLD1的迁移及侵袭能力,提示TM4SF1与结直肠癌侵袭转移密切相关,为进一步阐明TM4SF1在结直肠癌转移中的作用及相关机制奠定基础。
Objective To construct a eukaryotic expression vector of transmembrane-4-L-six-family-1 （TM4SF1） gene and study its effect on the migration and invasion of colorectal cancer cells. Methods A pair of specific primers of TM4SF1 gene （GenBank： BC034145.1） was used to acquire the open reading frame of TM4SF1 by RT-PCR. The amplified sequence was ligated to a PEZ-M29 vector, which, after identification, was transiently transfected in colorectal cancer cell lines HCT116 and DLD1. Western blotting and immunocytochemistry were used to analyze the transfection efficiency, and scratch and Transwell tests were performed to analyze the changes in the migration and invasion of HCT116 and DLD1 cells after transfection. Results Cell scratch and Transwell assays revealed that transfection with the recombinant plasmid, PEZ-M29/TM4SF1, caused up-regulated expression of TM4SF1 and promoted the migration and invasion of HCT116 and DLD1 cells. Conclusion Our results demonstrated that TM4SF1 is closely related to the invasion and metastasis of colorectal cancer cells in vitro.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2014年第6期847-851,共5页
Journal of Southern Medical University
基金
国家自然科学基金(81272763)~~
关键词
TM4SF1
迁移
侵袭
transmembrane-4-L-six-family-1
migration
invasion
