Profiling and bioinformatics analysis of differentially expressed circular RNAs in human intervertebral disc degeneration

Abstract

The functional changes of nucleus pulposus (NP) cells are considered to be the initiating factors of intervertebral disc degeneration (IDD), and the differentially expressed circRNAs in NP cells may play an important role in the process of IDD. To identify circular RNAs (circRNAs) associated with human IDD, we isolated the NP cells from human degenerated and non-degenerated intervertebral disc and identified NP cells by microscopy and cell proliferation. CircRNA microarray expression profiles were obtained from NP cells of degenerated and non-degenerated intervertebral disc and further validated by quantitative reverse transcription PCR (qRT-PCR). The expression data were analyzed by bioinformatics. Microarray analysis identified 7294 circRNAs differentially expressed in degenerated human IDD NP cells. Among them, 3724 circRNAs were up-regulated and 3570 circRNAs were down-regulated by more than 2 folds. After validating by qRT-PCR, we predicted the possible miRNAs of the top dysregulated circRNAs using TargetScan, and miRanda. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the most modulated circRNAs regulate the viability, degradation, apoptosis and oxidative stress in NP cells, and the possible mechanism underlying IDD was discussed. These results revealed that circRNAs may play a role in IDD and might be a promising candidate molecular target for gene therapy.

CircGLCE alleviates intervertebral disc degeneration by regulating apoptosis and matrix degradation through the targeting of miR-587/STAP1

“…25 To explore the role of circRNAs in IDD, a differential expression profile was produced and was analysed with a bioinformatic approach; the data revealed a promising candidate, CircGLCE. 26 In vivo experiments were performed using a rat tail model, which provides readily detected intervertebral disc degeneration through mechanical injury. 27 CircGLCE knockout and mutant rat models enabled the investigation of the effects of eliminating CircGLCE.…”

Gene expression profile of lipopolysaccharide‑induced apoptosis of nucleus pulposus cells reversed by syringic acid

“…Cells and cell culture. Human nP cells which had previously been isolated from nP tissues of patients with lumbar fracture (AO classification, B1) were kindly provided by Dr. Guo (Shanghai changzheng Hospital, Shanghai, china) (11). nP cells were cultured in dMeM/F12 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and 1% penicillin-streptomycin (Gibco; Thermo Fisher Scientific, Inc.) at 37˚C in an incubator in an atmosphere of 5% CO 2 .…”

Circular RNAs in nucleus pulposus cell function and intervertebral disc degeneration

“…The top ten differentially expressed circRNAs were validated by RT‐qPCR. Pathway analysis of the predicted miRNA targets of differentially expressed circRNAs indicated that cell cycle regulation, ubiquitin‐mediated proteolysis and ECM‐receptor interactions in NP cells are implicated in IDD development . In another study, Song and colleagues performed circRNA microarray with four NP specimens from normal and four from IDD subjects.…”