MiR-26a-5p Heightens Breast Cancer Cell Sensitivity to Paclitaxel via Targeting Flap Endonuclease 1

Ann Clin Lab Sci. 2023 Jan;53(1):116-125.

Abstract

Objective: Flap endonuclease 1 (FEN1) has been confirmed to involve the drug resistance of multiple cancers including breast cancer. However, the effect of miRNA-mediated FEN1 on breast cancer cell resistance is still ambiguous and needs further research.

Methods: Firstly, we used GEPIA2 to predict the FEN1 expression in breast cancer. Next, we used quantitative real-time polymerase chain reaction (qRT-PCR) and western blot to evaluate the FEN1 level of cells. After parental cells or MDA-MB-231-paclitaxel (PTX) cells being transfected with or without siFEN1, the apoptosis, migration, and protein levels of FEN1, Bcl-2, and resistance-related genes were examined by flow cytometry, wound healing assay, and western blot, respectively. Then, the putative miRNA targeting FEN1 was predicted using StarBase V3.0, and further confirmed by qRT-PCR. The targeted binding of FEN1 to miR-26a-5p was detected by dual-luciferase reporter assay. After parental cells or MDA-MB-231-PTX cells being transfected with or without miR-26a-5p mimic, the apoptosis, migration, and protein levels of FEN1, Bcl-2, and resistance-related genes were tested again.

Results: FEN1 expression was enhanced in breast cancer and MDA-MB-231-PTX cells. The combined application of FEN1 knockdown and PTX enhanced apoptosis in MDA-MB-231-PTX cells but suppressed cell migration and expressions of FEN1, Bcl-2, and resistance-related genes. Then, we confirmed that FEN1 was targeted by miR-26a-5p. The combined application of miR-26a-5p mimic and PTX largely facilitated apoptosis in MDA-MB-231-PTX cells but restrained cell migration and expressions of FEN1, Bcl-2, and resistance-related genes.

Conclusion: MiR-26a-5p contributes to the sensitivity of breast cancer cells to paclitaxel via restraining FEN1.

Keywords: breast cancer; flap endonuclease 1; miR-26a-5p; paclitaxel.

MeSH terms

  • Apoptosis / genetics
  • Breast Neoplasms* / drug therapy
  • Breast Neoplasms* / genetics
  • Breast Neoplasms* / metabolism
  • Cell Line, Tumor
  • Cell Proliferation / genetics
  • Female
  • Flap Endonucleases / genetics
  • Humans
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Paclitaxel / pharmacology
  • Proto-Oncogene Proteins c-bcl-2 / genetics

Substances

  • Paclitaxel
  • Flap Endonucleases
  • MicroRNAs
  • Proto-Oncogene Proteins c-bcl-2