Sun Lingling, Huan Pin, Wang Hongxia, Liu Fengsong, Liu Baozhong
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao, 266071, China.
Mol Biol Rep. 2014 May;41(5):2757-65. doi: 10.1007/s11033-014-3130-9. Epub 2014 Jan 28.
The epidermal growth factor receptor (EGFR) is an important receptor tyrosine kinase member in animals, which plays versatile functions in development, growth, tissue regeneration etc. Current knowledge on EGFR is poor in bivalve mollusks. In this study, we cloned and analyzed an EGFR gene from the Pacific oyster Crassostrea gigas (cgegfr). A 5,731 bp full-length cDNA of cgegfr was obtained, encoding a peptide with 1,494 amino acids which exhibited a typical EGFR structure, including an extracellular region, a single transmembrane region and an intracellular region. A conserved tyrosine kinase domain was predicted in the intracellular region, while the extracellular region responsible for ligand binding showed comparatively poor conservation. Expression analysis revealed that cgefgr was expressed widely in C. gigas tissues and a highest expression level was observed in adductor tissue. Expression of cgegfr was revealed to be up-regulated during wound healing of mantle, indicating that EGFR might function in the cell proliferation and migration during wound healing. Further functional analysis of cgegfr was conducted in mouse myoblast cell line C2C12, in which different parts of cgegfr were expressed and their effects were measured. The results revealed that cgegfr was able to accelerate cell proliferation of C2C12 cells and the transmembrane region was necessary for self-activation of truncated cgegfr. Our results would provide supports for further studies on the roles of cgegfr in development and growth in C. gigas.
表皮生长因子受体(EGFR)是动物体内一种重要的受体酪氨酸激酶成员,在发育、生长、组织再生等过程中发挥着多种功能。目前关于双壳贝类软体动物中EGFR的了解较少。在本研究中,我们克隆并分析了太平洋牡蛎(Crassostrea gigas)的一个EGFR基因(cgegfr)。获得了一个5731 bp的cgegfr全长cDNA,其编码一个含有1494个氨基酸的肽段,该肽段具有典型的EGFR结构,包括一个细胞外区域、一个单跨膜区域和一个细胞内区域。在细胞内区域预测到一个保守的酪氨酸激酶结构域,而负责配体结合的细胞外区域的保守性相对较差。表达分析表明,cgefgr在太平洋牡蛎组织中广泛表达,在内收肌组织中观察到最高表达水平。cgegfr的表达在套膜伤口愈合过程中被上调,表明EGFR可能在伤口愈合过程中的细胞增殖和迁移中发挥作用。在小鼠成肌细胞系C2C12中对cgegfr进行了进一步的功能分析,在该细胞系中表达了cgegfr的不同部分并测量了其作用。结果表明,cgegfr能够加速C2C12细胞的增殖,并且跨膜区域对于截短的cgegfr的自我激活是必需的。我们的结果将为进一步研究cgegfr在太平洋牡蛎发育和生长中的作用提供支持。