摘要
Kin17是一个与DNA复制、DNA修复有关的蛋白质,在人类的各种组织中表达均很低.目前乳腺上皮细胞生长增殖的分子机制尚未阐明.为了探讨Kin17与乳腺上皮细胞增殖的关系,检测了Kin17在不同增殖状况下的MCF-10A细胞中的表达情况,并把KIN17基因插入真核表达载体pcDNA3.1-(+)中,构建重组质粒pcDNA3.1-Kin17,通过转染MCF-10A细胞,检测Kin17的表达对MCF-10A细胞的增殖、DNA复制活性及信号分子表达的影响;同时,在转染Kin17特异性小干扰RNA(siRNA_Kin17)后,分析MCF-10A细胞的Kin17表达及细胞生长状况.实验结果显示,经高浓度血清刺激后,细胞中Kin17表达升高,而且生长越快的细胞,Kin17表达越强;转染重组质粒pcDNA3.1-Kin17明显提高了MCF-10A细胞中Kin17的表达,同时Kin17的上调表达促进了细胞的增殖速度与DNA复制活性,增强了cyclin D1的表达水平.当转染siRNA_Kin17时使Kin17含量下调,MCF-10A细胞生长速度的抑制不显著.实验结果表明,Kin17与乳腺上皮细胞的DNA复制及生长增殖密切相关.对Kin17在乳腺上皮细胞增殖中的作用及分子调控机制的深入探讨,将有助于揭示乳腺癌细胞快速增殖的潜在机制.
Kinl7 is participated in DNA replication and DNA repair and ubiquitously expressed at low levels in human tissues. To explore the connection of Kinl7 expression with breast epithelial cell proliferation, MCF-10A cells in different proliferation states were analyzed for Kinl7 expression. After transfected with a pcDNA3.1-Kinl7 plasmid, cell proliferaion, DNA replication and levels of signaling molecules expression in transfected MCF-10A cells were analyzed, and compared with cells transfected with siRNA_Kinl7. The results showed that Kinl7 overexpression increased in cell proliferation in high concentrations of serum, and the cell growth rate, DNA replication activity and cyclin D1 expression correlated with Kinl7 protein levels, whereas the knockdown of Kinl7 expression showed little effects. Our findings indicated that Kinl7 were related to DNA replication and proliferation of breast epithelial cells. Further studies about the roles of Kinl7 in breast epithelial cells is required for better understanding the underlying mechanisms.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2013年第5期457-462,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金(No.81201831)
广东省自然科学基金(No.S2012040006310)
广东省医学科学技术研究基金项目(No.B2012266)~~
关键词
Kin17
细胞增殖
乳腺上皮细胞
真核表达载体
Kinl7
cell proliferation
breast epithelial cell
eukaryotic expression vector
