储存红细胞中circRNA变化对泛素介导蛋白水解通路调控的预测分析*

doi:10.3969/j.issn.1671-2587.2021.04.004

临床输血与检验 ›› 2021, Vol. 23 ›› Issue (4): 420-427.DOI: 10.3969/j.issn.1671-2587.2021.04.004

• 储存血细胞中非编码RNA专题 • 上一篇下一篇

储存红细胞中circRNA变化对泛素介导蛋白水解通路调控的预测分析^*

张怡宇, 黄国清, 张勇刚, 苑召虎, 王强, 陈小洁, 黄建云, 李楠, 魏亚明

518000 深圳市龙华区中心医院（张怡宇,黄国清 ,张勇刚,王强）; 华南理工大学附属第二医院（广州市第一人民医院）,广东省精准输血技术工程研究中心（苑召虎,陈小洁,黄建云,李楠,魏亚明）

收稿日期:2021-04-12 发布日期:2021-08-18
通讯作者: 魏亚明,男,研究员,博士生导师,主要从事干细胞治疗与输血免疫学方面研究,（E-mail）eywym@scut.edu.cn。
作者简介:张怡宇（1988-）,女,广东深圳人,主管技师,硕士,主要从事输血免疫及分子生物学方面研究,（E-mail）273801816 @qq.com。并列第一作者：黄国清（1970-）,男,广东深圳人,主任技师,本科,主要从事输血方向研究,（E-mail）522415654@qq.com。
基金资助:
*本课题受广州市科技计划项目（No.201904010027）,广州市临床重大项目（No.2019ZD18）,国家自然科学基金青年项目（No.81801474）,深圳市科技计划项目（No.JCYJ20180306172502097）和深圳市龙华区科技创新项目（No.2020033）资助

Regulation of Ubiquitin Mediated Proteolytic Pathway: Predictive Analysis of CircRNA Changes in Stored Red Blood Cells

ZHANG Yi-yu, HUANG Guo-qing, ZHANG Yong-gang, et al

Department of Blood Transfusion, Shenzhen Longhua District Central Hospital, Shenzhen 518000

Received:2021-04-12 Published:2021-08-18

摘要/Abstract

摘要： 目的探讨储存红细胞中circRNA对泛素蛋白水解通路调节的可能机制及其在红细胞储存损伤中可能发挥的作用的预测分析。方法健康无偿献血者3（人）份血液制备的悬浮红细胞,过滤白细胞后平均分为新鲜组（0 d组）,4 ℃储存20 d红细胞组（20 d组）,4 ℃储存40 d红细胞组（40 d组）;对3组标本用高通量测序检测,然后做KEGG和GO分析,选取母基因在泛素介导的蛋白水解通路中的circRNA做PCR验证,用circBase和mirTarBase完成数据库注释与靶向预测,并将miRNA可能作用的mRNA也进行了关联分析。用circRNA interactome预测circRNA的RBP结合蛋白。结果与泛素蛋白水解调控相关的hsa_circ_0036044、hsa_circ_0024173、hsa-miR-4530表达量,20 d组均高于0 d组,差异有统计学意义（P<0.05）,20 d组的hsa_circ_0035761和TRAF2表达量低于0 d组,差异有统计学意义（P<0.05）;40 d组hsa_circ_0002502表达量低于0 d 组,且差异有统计学意义（P<0.05）;40 d 组的hsa_circ_0024173和hsa-miR-4530表达量低于与20 d 组,且差异有统计学意义（P<0.05）,40 d组的hsa_circ_0028196、hsa_circ_0035761、TRAF2表达量高于20 d组,且差异有统计学意义（P<0.05）。进一步通过生物信息学分析预测显示hsa_circ_0035761参与抗凋亡生理过程。结论本研究发现了多个体外储存红细胞中与泛素介导的蛋白水解通路的circRNA表达呈差异变化,发现和验证了hsa_circ_0035761—hsa-miR-4530—TRAF2轴,并揭示了hsa_circ_0035761还具有抗凋亡作用。与多种RNA结合蛋白结合能力,可能是这些circRNA表达量升高主要机制。

关键词: 红细胞储存损伤, 环状RNA分子, 小RNA分子, RNA结合蛋白, 凋亡

Abstract: Objective To investigate potential mechanism of circRNA regulating ubiquitin proteolytic pathway in stored red blood cells （RBCs） and its possible role in RBCs storage lesion. Methods The suspended RBCs （SRBCs） were collected from three healthy donors. After white blood cell （WBC） filtration,SRBCs were divided into fresh group （0-Day）group,stored RBCs for 20 days （20-Day） group and stored RBCs for 40 days（40-Day） group at 4℃. After high-throughput sequencing,Kyoto Encyclopedia of Genes and Genomes（KEGG）and Gene ontology （GO） analysis were performed. Selected circRNAs of parental gene in ubiquitin mediated proteolysis were verified by PCR. These circRNAs were annotated and targeted prediction by circBase database and mirTarBase. Then,analysis of miRNA-mRNA interactions was also performed. CircRNA interactome software was used to predict RNA-binding proteins（RBPs）on circRNAs. Results Compared to fresh group,the expression of hsa_circ_0036044,hsa_circ_0024173 and hsa-miR-4530 increased in 20-Day group,with a statistically significant（P<0.05）; while the expression of hsa_circ_0035761 and TRAF2 decreased, with a statistically significant（P<0.05）. Compared to fresh group,hsa_circ_0002502 decreased in 40-Day group,with a statistically significant（P<0.05）. Compared to the 20-Day group,hsa_circ_0024173 and hsa-miR-4530 decreased in 40-Day group,with a statistically significant（P<0.05）; while the expression of hsa_circ_0028196,hsa_circ_0035761 and TRAF2 increased,with a statistically significant （P<0.05）. Bioinformatics analysis showed that hsa_circ_0035761 might be involved in physiological processes mediate anti-apoptosis. Conclusion We found that multiple circRNA expressions were difference changes in ubiquitin-mediated proteolytic pathway,and also constructed the hsa_circ_0035761—hsa-miR-4530—TRAF2 axis. It revealed that hsa_circ_0035761 might have an anti-apoptotic effect on physiological processes. These circRNAs had ability to interact with many RNA-binding proteins,and this might be the main mechanism for high level expression of these circRNAs.

Key words: RBC storage lesion, CircRNA, miRNA, RBP, Apoptosis

中图分类号:

张怡宇, 黄国清, 张勇刚, 苑召虎, 王强, 陈小洁, 黄建云, 李楠, 魏亚明. 储存红细胞中circRNA变化对泛素介导蛋白水解通路调控的预测分析^*[J]. 临床输血与检验, 2021, 23(4): 420-427.

ZHANG Yi-yu, HUANG Guo-qing, ZHANG Yong-gang, et al. Regulation of Ubiquitin Mediated Proteolytic Pathway: Predictive Analysis of CircRNA Changes in Stored Red Blood Cells[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2021, 23(4): 420-427.

参考文献

[1] HERSHKO A.Some lessons from my work on the biochemistry of the ubiquitin system[J].J Biol Chem,2009,284(16):10291-10295.
[2] KHANDROS E,THOM C S,D'SOUZA J,et al.Integrated protein quality-control pathways regulate free α-globin in murine β-thalassemia[J].Blood,2012,119(22):5265-5275.
[3] MISCHO H E,CHUN Y,HARLEN K M,et al.Cell-cycle modulation of transcription termination factor Sen1[J].Mol Cell,2018,70(2):312-326.e7.
[4] DENG T,YAN G,SONG X,et al.Deubiquitylation and stabilization of p21 by USP11 is critical for cell-cycle progression and DNA damage responses[J].PNAS,2018,115(18):4678-4683.
[5] HERSHKO A.Ubiquitin-mediated protein degradation[J].J Biol Chem,1988,263(30):15237-15240.
[6] BONIZZATO A,GAFFO E,TE KRONNIE G,et al.CircRNAs in hematopoiesis and hematological malignancies[J].Blood Cancer J,2016,6(10):e483.
[7] SU Y,LV X,YIN W,et al.CircRNA Cdr1as functions as a competitive endogenous RNA to promote hepatocellular carcinoma progression[J].Aging (Albany NY),2019,11(19):8183-8203.
[8] XIONG D D,DANG Y W,LIN P,et al.A circRNA-miRNA-mRNA network identification for exploring underlying pathogenesis and therapy strategy of hepatocellular carcinoma[J].J Transl Med,2018,16(1): 220.
[9] ZHANG M, WANG T, XIAO G, et al.Large-Scale Profiling of RBP-circRNA Interactions from Public CLIP-Seq Datasets[J]. Genes (Basel), 2020;11(1):54.
[10] JECK W R,SHARPLESS N E.Detecting and characterizing circular RNAs[J].Nat Biotechnol,2014,32(5):453-461.
[11] 张怡宇,苑召虎,黄慧瑛,等.储存红细胞ATP代谢circRNA筛选及功能预测[J].中国输血杂志,2018,31(4):332-338.
[12] CHEN X,XIE X,XING Y,et al.MicroRNA dysregulation associated with red blood cell storage[J].Transfus Med Hemother,2018,45(6):397-402.
[13] ALHASAN A A,IZUOGU O G,AL-BALOOL H H,et al.Circular RNA enrichment in platelets is a signature of transcriptome degradation[J].Blood,2016,127(9): e1-e11.
[14] NICOLET B P,ENGELS S,AGLIALORO F,et al.Circular RNA expression in human hematopoietic cells is widespread and cell-type specific[J].Nucleic Acids Res,2018,46(16):8168-8180.
[15] TALHOUARNE G J S,GALL J G.7SL RNA in vertebrate red blood cells[J].RNA,2018,24(7):908-914.
[16] BORGHI A,VERSTREPEN L,BEYAERT R.TRAF2 multitasking in TNF receptor-induced signaling to NF-κB,MAP kinases and cell death[J].Biochem Pharmacol,2016,116:1-10.
[17] VAN EDEN M E,BYRD M P,SHERRILL K W,et al.Translation of cellular inhibitor of apoptosis protein 1 (c-IAP1) mRNA is IRES mediated and regulated during cell stress[J].RNA,2004,10(3):469-481.
[18] LI X,YANG Y,ASHWELL J D.TNF-RII and c-IAP1 mediate ubiquitination and degradation of TRAF2[J].Nature,2002,416(6878):345-347.
[19] ZHENG X Y,HUANG M G,XING L,et al.The circRNA circSEPT9 mediated by E2F1 and EIF4A3 facilitates the carcinogenesis and development of triple-negative breast cancer[J].Mol Cancer,2020,19(1):73.
[20] WANG R J,ZHANG S,CHEN X Y,et al.EIF4A3-induced circular RNA MMP9 (circMMP9) Acts as a sponge of miR-124 and promotes glioblastoma multiforme cell tumorigenesis[J].Mol Cancer,2018,17(1):166.

储存红细胞中circRNA变化对泛素介导蛋白水解通路调控的预测分析^*

Regulation of Ubiquitin Mediated Proteolytic Pathway: Predictive Analysis of CircRNA Changes in Stored Red Blood Cells

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