Analysis of differential expression LncRNA in intervertebral discs of spinal tuberculosis and screening of potential diagnostic markers

English Keywords:Spinal tuberculosis  LncRNA  Diagnostic markers

English Abstract:

  【Abstract】 Objectives: To detect the differentially expressed long non-coding RNA(LncRNA) and mRNA in intervertebral discs of patients with spinal tuberculosis(STB) by expression profile chip and filter LncRNA with the potential of becoming a diagnostic marker. Methods: 28 patients with STB(Test group) and 28 patients(Control group) with spinal degeneration were selected from the Department of Spinal Orthopaedics of General Hospital of Ningxia Medical University from September 2018 to December 2020. The diseased intervertebral discs of the two groups were collected and the total RNA was extracted. Three qualified samples from each group were detected by Arraystar Human LncRNA V5.0 chip and the fold change(FC) of each RNA was calculated by chip data processing. Among them, LncRNA and mRNA with FC≥2 were differentially expressed RNA(P<0.05). The differential mRNA adjacent to the differential LncRNA was regarded as the cis-target gene, and the protein that might be bound by LncRNA was regarded as the trans-target protein, and then the functions of mRNA, cis-target gene and trans-target protein were analyzed. The LncRNA whose cis-target gene may be related to the pathogenesis of STB was selected from the differential LncRNA and verified by RT-PCR in the remaining 25 samples to screen of LncRNA with the potential to become a diagnostic marker and then their functions were analyzed. Results: A total of 444 differential LncRNA and 186 differential mRNA were screened based on the difference of chip data between the two groups. The analysis of gene functional annotation showed that the cis-target genes of differential LncRNA were mainly involved in the regulation of biological processes such as transcription, translation, splicing, immune response, natural killer cell-mediated cytotoxicity and the transmission of signal pathways, while trans-target proteins were mainly involved in the regulation of gene expression processes such as RNA splicing, processing and transport. Differential mRNA was mainly involved in the regulation of transcription, translation, splicing, inflammation, immune response and other biological processes and the transmission of signal pathways. RT-PCR verification showed that there were significant down-regulated expressions of LINC01128-201 and Lnc-DNALI1-205 in STB patients, which were consistent with the results of microarray, and these two LncRNA may play an important role in the pathogenesis of STB by regulating the expression volume of ISG15, SNIP1 and HES4. Conclusions: There is a significant difference in the expression of LncRNA and mRNA between patients with STB and patients with spinal degeneration. The down-regulated expressions of LINC01128-201 and Lnc-DNALI1-205 may play an important role in the pathogenesis of STB and have the potential to become diagnostic markers.