储存红细胞中表达降低的circRNA以及其靶基因预测的研究*

doi:10.3969/j.issn.1671-2587.2021.04.005

临床输血与检验 ›› 2021, Vol. 23 ›› Issue (4): 428-434.DOI: 10.3969/j.issn.1671-2587.2021.04.005

• 储存血细胞中非编码RNA专题 • 上一篇下一篇

储存红细胞中表达降低的circRNA以及其靶基因预测的研究^*

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

518000 深圳市龙华区中心医院输血科（张怡宇,黄国清,王强,张勇刚）; 广州市第一人民医院输血科,广东省精准输血工程技术中心（陈小洁,黄建云,李楠,魏亚明）

收稿日期:2020-04-16 发布日期:2021-08-18
通讯作者: 魏亚明,男,主任技师,博士研究生导师,（E-mail）eywym@scut.edu.cn。
作者简介:张怡宇（1988-）,女,主管技师,硕士,主要从事输血医学方面研究,（E-mail）273801816@qq.com。
基金资助:
*本课题受广州市科技计划项目（No.201904010027）资助

Study on Decreased Expression of CircRNAs in Stored Erythrocytes and Their Target Genes Prediction

ZHANG Yi-yu, HUANG Guo-qing, WANG Qiang, et al

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

Received:2020-04-16 Published:2021-08-18

摘要/Abstract

摘要： 目的探讨储存红细胞中表达降低的circRNA靶基因预测及其与储存损伤的关系。方法 3（人）份血样来自健康无偿献血者血液制备的悬浮红细胞,滤除白细胞后均分为新鲜组（0 d）,4 ℃储存20 d红细胞组（储存组）;对2组标本用高通量测序检测,选取下降明显的10个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,以此筛选到可能与储存损伤密切相关的circRNA。结果 20 d与0 d组对比,hsa_circ_0005413、hsa_circ_0008937、hsa_circ_0040340、hsa_circ_0003126、hsa_circ_0007995、hsa_circ_0023937、novel_circ_0000402、hsa_circ_0028281表达量显著下降（P<0.05）;hsa-miR-7160-5p、hsa-miR-122-5p、hsa-miR-4534的表达量显著上升（P<0.05）;CASP8、CDK4、TMOD3显著下降（P<0.05）。生物信息学分析显示这些circRNA的靶基因参与红细胞凋亡与抗凋亡、红细胞膜的稳定、红细胞氧化损伤、铁代谢等生理过程。结论本研究构建了几个差异表达的circRNA-miRNA-mRNA网络,发现hsa_circ_0005413-hsa-miR-7160-5p-CASP8、hsa_circ_0040340-hsa-miR-122-5p-CDK4、hsa_circ_0007995-hsa-miR-4534-TMOD3可能参与了红细胞储存损伤的生物学过程。

关键词: 红细胞储存损伤, 环状RNA分子, 小分子RNA, 凋亡 CASP8

Abstract: Objective To investigate the relationship between significant decreased expression of circRNAs and its predicted target genes in erythrocyte storage lesion. Methods Whole blood collected from healthy donors is separated to suspended red blood cells after leukocyte filtration. The samples were divided into fresh groups（0 day）and storage groups（4℃ for 20 days）. The two groups of specimens were tested by high-throughput sequencing,and 10 down-regulated circRNAs were selected for PCR verification. CircBase and mirTarBase were used to database annotation and target gene prediction,and the association analysis of mRNAs which could be effected by miRNAs were also be done. Storage lesion-related circRNAs could be screened out through methods above. Results Compared to the fresh groups,the expression of hsa_circ_0005413,hsa_circ_0008937,hsa_circ_0040340,hsa_circ_0003126,hsa_circ_0007995,hsa_circ_0023937,
novel_circ_0000402,hsa_circ_0028281 significantly decreased in storage groups（P<0.05）,while the expression of hsa-miR-7160-5p,hsa-miR-122-5p and hsa-miR-4534 significantly increased（P<0.05）and the expression of CASP8,CDK4,and TMOD3 decreased significantly（P<0.05）. Bioinformatics analysis shows that these circRNA target genes are involved in apoptosis and anti-apoptosis,erythrocyte membrane stability,oxidative damage,iron metabolism and other physiological processes. Conclusion We constructed several differentially expressed circRNA-miRNA-mRNA triple network,and found that hsa_circ_0005413-hsa-miR-7160-5p-CASP8,hsa_circ_0040340-hsa-miR-122-5p-CDK4,hsa_circ_0007995-hsa-miR-4534-TMOD3 axis might participate in the biological process of erythrocyte storage lesion.

Key words: RBC storage lesion, CircRNA, miRNA, Apoptosis, CASP8

中图分类号:

R457

张怡宇, 黄国清, 王强, 张勇刚, 苑召虎, 陈小洁, 黄建云, 李楠, 魏亚明. 储存红细胞中表达降低的circRNA以及其靶基因预测的研究^*[J]. 临床输血与检验, 2021, 23(4): 428-434.

ZHANG Yi-yu, HUANG Guo-qing, WANG Qiang, et al. Study on Decreased Expression of CircRNAs in Stored Erythrocytes and Their Target Genes Prediction[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2021, 23(4): 428-434.

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储存红细胞中表达降低的circRNA以及其靶基因预测的研究^*

Study on Decreased Expression of CircRNAs in Stored Erythrocytes and Their Target Genes Prediction

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