miR-1 Targeted Downregulation of Bcl-2 Increases Chemosensitivity of Lung Cancer Cells

Genet Test Mol Biomarkers. 2021 Aug;25(8):540-545. doi: 10.1089/gtmb.2021.0009.

Abstract

Objective: To investigate the expression of B cell lymphoma-2 (Bcl-2) in lung cancer cells and the effect of the miR-1/Bcl-2 axis on the chemosensitivity of lung cancer. Materials and Methods: Real-time quantitative PCR and western blotting were used to detect the expression of Bcl-2 in human embryonic lung fibroblasts and lung cancer cells. The effects of siRNA directed against Bcl-2, in lung cancer tissue samples was detected by immunohistochemistry; these results were used to develop prognostic models. Bioinformatic analyses, dual luciferase reporter gene technology, and western blotting technology were used to explore the targeted regulation of miR-1 on bcl-2. The effect of miR-1 on the chemosensitivity of lung cancer cells was measured using the MTT assay. Results: Compared with human embryonic lung fibroblasts, Bcl-2 was highly expressed in the lung cancer cells, especially in H460 cells. After silencing Bcl-2 with siRNA, the sensitivity of the cells to cisplatin (CDDP) increased. Immunohistochemical results and prognostic analysis revealed that high Bcl-2 expression in lung cancer tissues was negatively correlated with prognosis of lung cancer patients; A dual luciferase reporter assay combined with western blotting confirmed that miR-1 can bind to the Bcl-23' UTR region and regulate its expression. Overexpression of miR-1 in lung cancer cells (H460 and A549) increased the sensitivity of these cells to CDDP. Conclusion: Bcl-2 is upregulated in lung cancer cells, which is negatively correlated with the patient prognosis. miR-1 affects the chemosensitivity of lung cancer cells by targeting Bcl-2. These data should provide a theoretical basis for refining the molecular mechanisms of chemoresistance in lung cancer.

Keywords: Bcl-2; chemosensitivity; lung cancer; miR-1; prognosis.

MeSH terms

  • Antineoplastic Agents / pharmacology
  • Apoptosis / drug effects
  • Carcinoma, Non-Small-Cell Lung / genetics
  • Cell Line, Tumor
  • China
  • Cisplatin / pharmacology
  • Cisplatin / therapeutic use
  • Drug Resistance, Neoplasm / genetics
  • Gene Expression / genetics
  • Gene Expression Profiling / methods
  • Gene Expression Regulation, Neoplastic / genetics
  • Humans
  • Immunohistochemistry
  • Lung / metabolism
  • Lung / pathology
  • Lung Neoplasms / drug therapy
  • Lung Neoplasms / genetics*
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / genetics*
  • Transcriptome / genetics

Substances

  • Antineoplastic Agents
  • BCL2 protein, human
  • MIRN1 microRNA, human
  • MicroRNAs
  • Proto-Oncogene Proteins c-bcl-2
  • Cisplatin