Guo Zhi Liang, Liu Yuan Yuan, Gao Yu, Guan Xiu Mei, Li Hong, Cheng Min
Department of Orthopedic, No. 89 Hospital of Chinese PLA, Weifang, 261021, China. Email:
Stomatology Medical College, Weifang Medical University, Weifang 261053, Shandong, China.
Cell J. 2021 Jul;23(2):218-224. doi: 10.22074/cellj.2021.6832. Epub 2021 May 26.
The present study aimed to screen the differentially expressed (DE) circular RNAs (circ-RNAs) between lumbar intervertebral disc degeneration (IVDD) and normal tissues.
In this experimental study, microarray hybridization was performed to evaluate circ-RNA expression, and the DE circ-RNAs were confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). Host genes of DE circ-RNAs were predicted, and their functions were evaluated. Further, a competitive endogenesis (ce) RNA network among 4 DE circ-RNAs-miRNA-mRNA was constructed by Cytoscape.
A total of 2636 circ-RNAs were detected in all samples; among them, 89.23% were exonic circ-RNAs. There were 138 DE circ-RNAs, including 134 up-regulated circ-RNAs and 4 downregulated circ-RNAs in IVDD samples. qRT-PCR validation experiments showed that expression trends of hsa_circ_0003239, hsa_circ_0003162, hsa_circ_0005918, and hsa_circ_0005556 were in line with the microarray analysis results. Functional enrichment analysis showed that host genes of DE circ-RNAs significantly disturbed pathways of regulation of actin cytoskeleton, propanoate metabolism, and ErbB signaling pathway. The four DE circ-RNAs related ceRNA network was constructed.
Our results revealed that circ-RNAs can function as miRNA sponges and regulate parent gene expression to affect IVDD.
本研究旨在筛选腰椎间盘退变(IVDD)组织与正常组织之间差异表达的环状RNA(circ-RNA)。
在本实验研究中,进行微阵列杂交以评估circ-RNA表达,并通过定量实时聚合酶链反应(qRT-PCR)确认差异表达的circ-RNA。预测差异表达circ-RNA的宿主基因,并评估其功能。此外,利用Cytoscape构建了4种差异表达circ-RNA-微小RNA(miRNA)-信使核糖核酸(mRNA)之间的竞争性内源性(ce)RNA网络。
在所有样本中共检测到2636种circ-RNA;其中,89.23%为外显子circ-RNA。在IVDD样本中有138种差异表达的circ-RNA,包括134种上调的circ-RNA和4种下调的circ-RNA。qRT-PCR验证实验表明,hsa_circ_0003239、hsa_circ_0003162、hsa_circ_0005918和hsa_circ_0005556的表达趋势与微阵列分析结果一致。功能富集分析表明,差异表达circ-RNA的宿主基因显著干扰肌动蛋白细胞骨架调节、丙酸代谢和表皮生长因子受体(ErbB)信号通路。构建了4种差异表达circ-RNA相关的ceRNA网络。
我们的结果表明,circ-RNA可作为miRNA海绵发挥作用,并调节亲本基因表达以影响IVDD。