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

doi:10.3969/j.issn.1671-2587.2021.04.004

JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE ›› 2021, Vol. 23 ›› Issue (4): 420-427.DOI: 10.3969/j.issn.1671-2587.2021.04.004

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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

Abstract

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

CLC Number:

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.

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

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