基础医学与临床 ›› 2016, Vol. 36 ›› Issue (6): 767-771.

• 研究论文 • 上一篇    下一篇

过表达TRIM69抑制细胞内源c-Jun蛋白的表达

王任先1,李凯2,姚荣彦2,缪时英3,王琳芳3,宋伟4   

  1. 1. 北京积水潭医院
    2. 中国医学科学院
    3. 中国医学科学院 基础医学研究所
    4. 中国医学科学院基础医学研究所,北京协和医学院基础学院,医学分子生物学国家重点实验室,生物化学与分子生物学系
  • 收稿日期:2016-03-17 修回日期:2016-04-20 出版日期:2016-06-05 发布日期:2016-05-27
  • 通讯作者: 宋伟 E-mail:roy_sw0925@yahoo.com.cn
  • 基金资助:
    国家自然科学基金;北京市优秀人才培养计划项目;国家重点实验室专项基金

Over expression of TRIM69 inhibits the expression of endogenous c-Jun

  • Received:2016-03-17 Revised:2016-04-20 Online:2016-06-05 Published:2016-05-27
  • Contact: Wei SONG E-mail:roy_sw0925@yahoo.com.cn
  • Supported by:
    the National Natural Science Foundation of China

摘要: 目的 探索TRIM69负调控AP1信号通路及抑制内源c-Jun蛋白表达的分子机制。方法 在子宫颈癌细胞HeLa细胞或人胚肾细胞HEK293T细胞共转染荧光素酶报告基因质粒和空载体或TRIM69,共转空载体组作为对照,通过双荧光素酶报告基因实验检测了TRIM69对8条关键信号通路的影响;在HEK293T细胞过表达TRIM69全长及各个结构域的截短体,通过Western blot实验检测了TRIM69抑制细胞内源c-Jun蛋白表达的关键结构域;在HEK293T细胞过表达TRIM69,利用cycloheximide (CHX)、MG132或chloroquine分别处理细胞,通过蛋白稳定性实验探讨了TRIM69负调控内源c-Jun蛋白表达的分子机制。结果 TRIM69可以特异负调控AP1信号通路(P<0.05),并且TRIM69对内源c-Jun蛋白表达的抑制作用依赖于其RBCC结构域(P<0.05),进一步研究发现TRIM69通过蛋白酶体途径影响内源c-Jun蛋白的稳定性(P<0.05)。结论 TRIM69负调控AP1信号通路并通过蛋白酶体途径影响内源c-Jun蛋白的降解。

关键词: TRIM69, AP1信号通路, 转录因子c-Jun

Abstract: Objective Explore the mechanism of TRIM69 inhibits AP1 pathway and down regulates the expression of c-Jun. Methods Co-transfect luciferase reporter plasmids and empty vector or TRIM69 plasmid in HeLa cells or HEK293T cells and make the empty vector group as control, dual-luciferase reporter assay system was used to check the effect of TRIM69 on eight important pathways; over express all length or the deletion mutants of TRIM69 in HEK293T cells and western blot was used to examine the key domain which inhibits the expression of c-Jun; over express TRIM69 in HEK293T cells and then cells were treated with cycloheximide (CHX), MG132 or chloroquine, CHX-chase assay was used to investigate the mechanism of TRIM69 down regulates the expression of c-Jun. Results TRIM69 could inhibit AP1 pathway and down regulate the expression of c-Jun which depends on its RBCC domain (P<0.05). TRIM69 negatively regulates the expression of c-Jun through proteasome degradation (P<0.05). Conclusions TRIM69 could down regulate AP1 pathway and inhibit the expression of c-Jun which depends on proteasome pathway.

Key words: TRIM69, AP1 pathway, c-Jun

中图分类号: