农业生物技术学报2024，Vol.32Issue(9)：2088-2099,12.DOI:10.3969/j.issn.1674-7968.2024.09.012
MDCK细胞成瘤性相关环状RNA的筛选与验证
Screening and Validation of Tumorigenicity-related CircRNAs in MDCK Cells
摘要
Abstract
In the preparation of animal vaccines such as avian influenza using the canine(Canis lupus familiaris)kidney cell line Madin-Darby canine kidney(MDCK),it was found that the MDCK cell line has tumorigenicity.In this study,highly tumorigenic and non-tumorigenic MDCK cells were selected for RNA-seq high-throughput sequencing.The differentially expressed circular RNA(circRNA)were obtained,and their source genes were analyzed by GO and KEGG.The StarBase and mirTarBase databases were used to predict target microRNA(miRNA)and mRNA of tumorigenic circRNA.Total RNA of tumorigenic and non-tumorigenic MDCK cells were extracted using TRIzol method,and the expression levels of 6 differential circRNAs were verified by qPCR.The results showed that 3 491 circRNAs were identified with the length of 200～800 nt.Among the screened out 27 differentially expressed circRNAs,18 were up-regulated and 9 were down-regulated.GO,KEGG and Reactome enrichment analysis revealed that differentially expressed circRNAs were associated with ATP activity and amino acid metabolism-related enzymes,linked to adhesion,migration,and various metabolic pathways.The network construction of competing endogenous RNA(ceRNA)illustrated 4 key circRNA-miRNA-mRNA regulatory network related to MDCK cell tumorigenicity,novel_circ_002222/novel-m0145-3p/dynactin 1(Dctn1),novel_circ_002222/miR-197-x1/Dctn1,novel_circ_001205/novel-m0580-5p/annexin A11(ANXA11),and novel_circ_003257/novel-m0675-3p/RNA-binding protein 48(RBM48).The above results indicate that differentially expressed circRNAs related to tumorigenicity in MDCK cells are mainly enriched in transmembrane material transport and energy metabolism-related pathways,which might play a role in energy uptake and metabolism in MDCK cells.This study provides a research foundation and technical support for revealing the mechanism of circRNA involvement in MDCK cell tumorigenesis and establishing genetically engineered non-tumorigenic MDCK cell lines.关键词
MDCK(Madin-Darby canine kidney)/环状RNA(circRNA)/成瘤性Key words
Madin-Darby canine kidney(MDCK)/Circular RNA(circRNA)/Tumorigenicity分类
农业科技引用本文复制引用
杨迪,石嘉琛,黄玲巍,王家敏,马玉梅,乔自林,崔燕..MDCK细胞成瘤性相关环状RNA的筛选与验证[J].农业生物技术学报,2024,32(9):2088-2099,12.基金项目
甘肃省教育厅高校教师创新基金(2023B-058) （2023B-058）
中央高校基本科研业务费资金专项(31920240063) （31920240063）
甘肃省科技计划(23YFFA0071) （23YFFA0071）
