宫颈癌细胞中与UTF1相互作用的蛋白分析

Identification of proteins that interact with UTF1 in cervical cancer cell

目的探讨宫颈癌细胞中与转录辅激活因子(UTF1)相互作用的蛋白。方法构建稳定表达FLAG-HA双标签标记的UTF1蛋白的SiHa细胞株,利用FLAG-HA串联亲和纯化(TAP)双标签纯化实验,对目的条带进行质谱分析。结果成功构建稳定表达FLAG-HA双标签标记的UTF1细胞株。通过质谱分析得到UTF1相互作用的蛋白数据,结果显示UTF1捕获蛋白参与DNA修复、代谢、核糖体、细胞连接、细胞因子相互作用、吞噬等多种生物学过程,以及Jak-STAT、MAPK、mTOR、VEGF、 wnt等多条信号通路,并与系统性红斑狼疮、帕金森症、阿尔兹海默症、自身免疫性疾病等的发病相关。结论 UTF1捕获蛋白参与细胞多种生理及病理过程,为进一步了解UTF1在肿瘤发病机制中的作用提供了新的线索。

Objective To search for the proteins interacting with undifferentiated embryonic cell transcription factor 1(UTF1) in cervical cancer cells.Methods We constructed SiHa cell line which stably expressed the UTF1 protein labeled with FLAG-HA double tag.After FLAG-HA double-tag tandem affinity purification experiment.The protein composition of target band was determined using mass-spectrometry.Results The UTF1 cell line that stably expressed FLAG-HA double-tag was successfully constructed.Through mass-spectrometry analysis,we found that captured protein of UTF1 is involved in various biological processes such as DNA repair,metabolism,ribosome,cell conjunction,interacions among various cytokines,phagocytosis,and many signal pathways including Jak-STAT,MAPK,mTOR,VEGF,wnt.The captured protein was associated with the development of systemic lupus erythematosus,Parkinson's disease,Alzheimer's disease,and autoimmune diseases.Conclusion The captured protein of UTF1 is involved in a variety of physiological and pathological processes of the cells,providing new clues for further understanding of the role of UTF1 in the pathogenesis of tumors.