Hnrnpk基因在猪成肌细胞增殖分化过程中的表达及分子调控机制研究

项目名称： Hnrnpk基因在猪成肌细胞增殖分化过程中的表达及分子调控机制研究

项目编号： No.U1204326

项目类型： 联合基金项目

立项/批准年度： 2013

项目学科： 畜牧学与草地科学、兽医学、水产学

项目作者： 徐永杰

作者单位： 信阳师范学院

项目金额： 30万元

中文摘要： Hnrnpk是一个多功能蛋白，参与DNA修复、转录和翻译调控、RNA的剪切加工等生物学过程。以往研究表明，Hnrnpk可能通过多种途径对骨骼肌细胞增殖与分化起重要调控作用，但其作用机制及信号通路尚不清楚。我们前期蛋白质组学研究发现Hnrnpk在梅山猪不同发育时期骨骼肌中差异表达。基于此，本项目拟结合过表达、RNA干涉、细胞转染、免疫印记、免疫组化、免疫荧光、qRT-PCR、流式细胞、透射电镜等生物学手段，以猪成肌细胞为体外模型，以Hnrnpk调控转录因子c-Myc和酪氨酸磷酸化激酶c-Src的活性、ERK和c-Src阻断剂对Hnrnpk的影响及磷酸化Hnrnpk的亚细胞定位和作用为切入点，研究Hnrnpk基因对猪成肌细胞增殖分化的调控作用及信号通路。项目的研究结果不仅能为阐明骨骼肌发育和再生的复杂网络调节机制奠定基础，而且还可望为畜牧业提高肉产量和肉品质以及医学上控制肌肉再生提供依据。

中文关键词： 猪；成肌细胞；增殖分化；基因功能；调控

英文摘要： Hnrnpk （heterogeneous nuclear ribonucleoprotein K）is a multifunctional protein that participate in chromatin remodeling, transcription, RNA splicing, mRNA stability and translation. Previous studies showed that Hnrnpk may play an important role in skeletal muscle cells proliferation and differentiation, but how and why it has this effect remains unknown. Our former research found that Hnrnpk was differentially expressed in the porcine skeletal muscle of different development stages by proteome analysis. Based on this observation, the subject was propounded. To reveal molecular egulatory mechanisms and signal pathway of Hnrnpk gene in porcine skeletal myoblasts proliferation and differentiation, several technologys were used, such as overexpression, RNAi, cell transfection, immunoblotting, immunohistochemistry, immunofluorescent, qRT-PCR, flow cytometry and transmission electron microscope in this study. Experimental investigation mainly contained: Experimental investigation mainly contained: the regulation effect of Hnrnpk in the expression of transcription c-Myc and activity of tyrosine kinase c-Src; the subcellular localization and expression of phosphorylated Hnrnpk in porcine myoblasts; the effects of ERK and c-Src inhibitor for Hnrnpk. The project lays a foundation for illustrating the regulatory mechanism

英文关键词： Pig；Myoblast；Proliferation and differentiation；Gene function；Regulation