Investigation the immunotherapeutic potential of miR-4477a targeting PD-1/PD-L1 in breast cancer cell line using a CD8(+) co-culture model.

Tülüce Y., Köstekci S., Karakuş F., Keleş A. Y., Tunçyürekli M.

Molecular biology reports, cilt.52, sa.1, ss.326, 2025 (SCI-Expanded, Scopus)

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 52 Sayı: 1
  • Basım Tarihi: 2025
  • Doi Numarası: 10.1007/s11033-025-10435-0
  • Dergi Adı: Molecular biology reports
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, CAB Abstracts, Chemical Abstracts Core, Veterinary Science Database
  • Sayfa Sayıları: ss.326
  • Van Yüzüncü Yıl Üniversitesi Adresli: Evet

Özet

Abstract

Background: In the present study, we investigated the immunotherapeutic and anticancer activities of microRNA-4477a (miR-4477a) as a PD-L1 inhibitor in breast cancer cells (MCF-7).

Methods: To this end, a series of analytical procedures were conducted, including bioinformatic analysis, RT-PCR analysis, PD-L1 ELISA, in vitro co-culture analysis, cytotoxicity assays, cell migration assays, and colony formation assays, with the objective of determining the anticancer activity of the compound in question.

Results: The results demonstrated that miR-4477a can bind to three distinct regions of PD-L1 mRNA with high scores (94%, 88% and 80%), effectively targeting and suppressing the crucial regulatory pathways of cancer cells. In vitro studies demonstrated that a 25 nM dose of miR-4477a caused relatively high cytotoxicity in the MCF-7 cell line, suppressed PD-L1 gene expression, and decreased sPD-L1 protein levels, strongly inhibited cell migration, and significantly reduced colony formation. The in vitro co-culture analysis revealed that cancer cells were unable to evade the surveillance and cytotoxic activity of T cells (CD8+) due to the blockade of PD-L1 expression by miR-4477a.

Conclusions: In conclusion, miRNA-4477a has the capacity to regulate immune responses in breast cancer cells and may therefore be a promising candidate for use in cancer immunotherapy as a therapeutic agent.

Keywords: Breast cancer; Immune checkpoint; Immunotherapy; PD-L1; miRNA-4477a.