Liu Rui, Cheng Qi, Wang Xiudan, Chen Hao, Wang Weilin, Zhang Huan, Wang Lingling, Song Linsheng
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China.
School of Food Science and Technology, Dalian Polytechnic University, Dalian 116034, China.
Fish Shellfish Immunol. 2017 Feb;61:68-78. doi: 10.1016/j.fsi.2016.12.005. Epub 2016 Dec 8.
B-cell translocation gene 1 (BTG1) is a member of the anti-proliferative gene family, which plays important roles in regulation of cell cycle. In the present study, a B-cell translocation gene 1 molecule homologue (designed CgBTG1) are identified and characterized in oyster Crassostrea gigas. CgBTG1 contains a conserved BTG domain with Box A and Box B motifs, and it shares high similarities with both BTG1 and BTG2 proteins in vertebrates. CgBTG1 mRNA is predominantly expressed in hemocytes, and its expression level in hemocytes is significantly up-regulated at 6 h (5.40-fold, p < 0.01) post Vibrio splendidus stimulation. The apoptosis rate of oyster hemocytes is significantly decreased (p < 0.05) after CgBTG1 interfered by dsRNA (dsCgBTG1). This is indicated that CgBTG1 participated in the regulation of oyster hemocytes apoptosis. Furthermore, CgBTG1 could also induce the apoptosis of cancer cells (HeLa, A549 and BEL7402) in vitro. Compared with normal oysters, both vessel-like structures and muscle fibers in CgBTG1 interfered oysters are severely damaged after V. splendidus challenge in paraffin section, considering that CgBTG1 possessed an analogous feature of angiogenesis for maintenance of vessel-like structures in adductor muscle of oyster. The results suggests that CgBTG1 is a multi-functional molecule involved in the immune response of C. gigas against pathogen infection, which provides more clues for intensive studies of BTG family proteins in invertebrates.
B细胞易位基因1(BTG1)是抗增殖基因家族的成员,在细胞周期调控中发挥重要作用。在本研究中,在太平洋牡蛎中鉴定并表征了一种B细胞易位基因1分子同源物(命名为CgBTG1)。CgBTG1包含一个带有A框和B框基序的保守BTG结构域,并且与脊椎动物中的BTG1和BTG2蛋白具有高度相似性。CgBTG1 mRNA主要在血细胞中表达,在灿烂弧菌刺激后6小时,其在血细胞中的表达水平显著上调(5.40倍,p < 0.01)。在用dsRNA(dsCgBTG1)干扰CgBTG1后,牡蛎血细胞的凋亡率显著降低(p < 0.05)。这表明CgBTG1参与了牡蛎血细胞凋亡的调控。此外,CgBTG1在体外也能诱导癌细胞(HeLa、A549和BEL7402)凋亡。与正常牡蛎相比,在石蜡切片中,经灿烂弧菌攻击后,干扰CgBTG1的牡蛎中的血管样结构和肌纤维均受到严重损伤,考虑到CgBTG1具有类似血管生成的特征,以维持牡蛎闭壳肌中的血管样结构。结果表明,CgBTG1是一种参与太平洋牡蛎对抗病原体感染免疫反应的多功能分子,这为深入研究无脊椎动物中的BTG家族蛋白提供了更多线索。