The role of differentially expressed miR-660 in peripheral blood lymphocytes of patients with pulmonary tuberculosis

Biomarkers. 2023 Dec;28(5):409-415. doi: 10.1080/1354750X.2023.2198166. Epub 2023 Apr 9.

Abstract

Objective: This study aimed to investigate the significance of miRNA expression levels in peripheral blood lymphocytes of patients clinically diagnosed with pulmonary tuberculosis.

Method: Pulmonary tuberculosis-related datasets in the Gene Expression Omnibus (GEO) database were analyzed, and DE-miRNAs were screened for Gene Ontology (GO) analysis and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway enrichment to construct a DE-miRNA-DE-mRNA network. The peripheral blood lymphocytes of 10 patients with pulmonary tuberculosis, 10 patients with rifampicin-resistant tuberculosis and 10 healthy volunteers were selected for validation of RNA expression levels. qRT-PCR was done to verify the expression of DE-miRNA, and western blotting was done to check the expression levels of genes of associated pathways.

Results: Differential expression of miR-660 was found in pulmonary tuberculosis through data analysis and literature mining. The differential expression was also confirmed by qRT-PCR in samples from patients and healthy controls. The expression of miR-660 was significantly upregulated (p < 0.01) in patients with pulmonary tuberculosis and rifampicin-resistant pulmonary tuberculosis compared with the healthy controls. According to western blotting results, the expression levels of P-NF-κB and AKT in patients with pulmonary tuberculosis and NF-κB, P-NF-κB, AKT and p-AKT in patients with rifampicin-resistant tuberculosis were significantly upregulated (p < 0.01).

Conclusion: The high expression levels of miR-660 may activate the AKT/NF-κB signalling pathway and has the potential to serve as a potential biomarker for the diagnosis of pulmonary tuberculosis.

Keywords: Tuberculosis; clinical diagnostic value; differentially expressed genes; miR-660.

MeSH terms

  • Gene Expression Profiling
  • Humans
  • Lymphocytes / metabolism
  • MicroRNAs*
  • NF-kappa B
  • Proto-Oncogene Proteins c-akt / genetics
  • Rifampin / pharmacology
  • Tuberculosis, Pulmonary* / diagnosis
  • Tuberculosis, Pulmonary* / genetics

Substances

  • NF-kappa B
  • Proto-Oncogene Proteins c-akt
  • Rifampin
  • MicroRNAs
  • MIRN660 microRNA, human