医学分子生物学杂志 ›› 2022, Vol. 19 ›› Issue (1): 20-25.doi: 10.3870/j.issn.1672-8009.2022.01.003

• 论著 • 上一篇    下一篇

自噬-溶酶体途径调控核仁小核糖核蛋白 RRP9 的蛋白质降解

  

  1. 首都医科大学基础医学院细胞生物学系 北京市, 100069
  • 出版日期:2022-01-31 发布日期:2022-02-25
  • 基金资助:
    国家自然科学基金 (No. 31971229), 蛋白质组学国家重点 实验室开放课题 (No. SKLP-O201901)

Autophagy-lysosomal Pathway Regulates the Protein Degradation of the Small Nucleolar Ribonucleoprotein RRP9

  1. Department of Cell Biology, School of Basic Medical Science, Capital Medical University, Beijing, 100069, China 
  • Online:2022-01-31 Published:2022-02-25

摘要: 目的 研究核仁小核糖核蛋白 RRP9 的蛋白降解机制。 方法 组织表达谱分析 RRP9 在小鼠和人 类正常组织中的种属特异性和在不同组织中的特异性表达, 并分析 RRP9 在人的正常结直肠组织和结直肠 癌组织中的表达差异; 使用放线菌酮 (cycloheximide, CHX) 抑制结直肠癌细胞蛋白合成来检测 RRP9 蛋白 半衰期; 抑制自噬-溶酶体途径后检测 RRP9 蛋白含量变化以明确自噬-溶酶体途径对 RRP9 蛋白水平的影 响; 最后通过免疫荧光和免疫共沉淀检测 LC3b 与 RRP9 之间的相互作用。 结果 RRP9 为长寿命蛋白; 抑 制自噬-溶酶体途径使 RRP9 蛋白水平升高; RRP9 与 LC3b 之间存在相互作用。 结论 自噬-溶酶体途径调 控 RRP9 蛋白的降解。

关键词: 结直肠癌, RRP9, 自噬-溶酶体途径

Abstract: Objective To study the protein degradation mechanism of the small nucleolar ribonucleoprotein RRP9. Methods Tissue expression profiling was used to analyze the species-specific and tissue-specific expression of RRP9 in normal tissues of mice and humans, and the differential expression of RRP9 in normal colorectal tissues and colorectal cancer tissues. Cycloheximide (CHX), a protein synthesis inhibitor, was used to detect the half-life of RRP9 in colorectal cancer cells. After inhibiting the autophagy-lysosome pathway, the expression of RRP9 was detected to understand the effect of autophagy-lysosomal pathway on RRP9. Finally, immunofluorescence and co-immunoprecipitation were used to detect the interaction between LC3b and RRP9. Results RRP9 is a long-lived protein. Inhibition of autophagy-lysosomal pathway increased the level of RRP9. There was an interaction between RRP9 protein and LC3b in colorectal cancer cells. Conclusion The autophagy-lysosomal pathway regulates the degradation of RRP9 protein.

Key words: colorectal cancer, RRP9, autophagy-lysosomal pathway

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