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日本三角涡虫体内Djcullin1基因和Djtinp1基因的相互作用研究
Study on the Interaction of Djcullin1 and Djtinp1 Genes in Planarian Dugesia japonica
张芬熙1,2, 董自梅1, 陈广文1*, 刘德增1
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DOI:10.11984/j.issn.1000-7083.20200247
作者单位:1. 河南师范大学生命科学学院, 河南新乡 453007;
2. 新乡医学院基础医学院, 河南新乡 453003
中文关键字:日本三角涡虫;Djcullin1基因;Djtinp1基因;RNA干扰;再生
英文关键字:Dugesia japonica; Djcullin1 gene; DjTINP1 gene; RNAi; negative feedback regulation
中文摘要:Cullin1蛋白是细胞周期进展的重要参与者,也是泛素连接酶E3的重要骨架蛋白。TGF-β信号通路在涡虫再生中发挥着重要作用,其下游分子TGF-β诱导核蛋白(Tinp1)也参与日本三角涡虫Dugesia japonica再生。虽然cullin1基因和tinp1基因在涡虫体内均呈阳性表达,但二者之间的关系还不清楚。本研究采用RNA干扰技术(RNAi)敲减日本三角涡虫体内Djcullin1基因的表达,然后运用整体原位杂交及Western Blotting技术检测基因敲减后TGF-β信号通路相关分子在涡虫体内的表达。结果显示,敲减日本三角涡虫Djcullin1基因可明显增强TGF-β信号通路相关分子Smad2/3及Smad4的表达,Tinp1的表达也明显增强。这些数据表明,在日本三角涡虫体内敲减Djcullin1基因可负性调控TGF-β信号通路和Djtinp1基因的表达。同时,敲减涡虫Djtinp1基因可增强Djcullin1基因及蛋白的表达,以上结果说明,这2个基因之间存在互相调节作用,Djtinp1基因反作用于Djcullin1基因的原因未知。
英文摘要:Cullin1 is a key participant in cell cycle progression and an important skeleton protein of ubiquitin ligases E3. TGF-β signaling pathway plays a critical role in planarian regeneration, and its downstream molecule TGF-β induces nuclear protein (Tinp1) also participates in the regeneration of Dugesia japonica. Although both cullin1 and tinp1 genes are expressed in planarians, the relationship of the two genes still remains unexplored. In this study, RNA interference (RNAi) technique was used to knockdown the Djcullin1 gene of D. japonica, and then whole-mount in situ hybridization and Western Blotting were used to detect the expression levels of TGF-β signaling pathway-related molecules. The results showed that knockdown of Djcullin1 gene significantly enhanced the expression of TGF-β pathway related molecules Smad2/3 and Smad4, the expression of Tinp1 was also significantly enhanced. Above indicated that the TGF-β signaling and Djtinp1 gene in planarians were negatively regulated by Djcullin1 gene. Meanwhile, knockdown of Djtinp1 could also increase Djcullin1 at transcriptional and protein levels. Therefore, this study preliminarily reveals that there was mutual regulation between genes Djcullin1 and Djtinp1 planarians.
2021,40(2): 142-149 收稿日期:2020-06-27
分类号:Q955
基金项目:国家自然科学基金项目(31570376;31471965;U1604173);河南省高校科技创新团队项目(18IRTSTHN022)
作者简介:张芬熙,博士研究生,研究方向:动物资源与保护,E-mail:mengfei2008lulu@126.com
*通信作者:陈广文,教授,博导,E-mail:chengw0183@sina.com
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