Tumor Cell-selective Synergism of TRAIL- and ATRA-induced Cytotoxicity in Breast Cancer Cells

Anticancer Res. 2018 May;38(5):2669-2682. doi: 10.21873/anticanres.12509.

Abstract

Background/aim: One of the major problems in breast cancer treatment is pharmacoresistance. Therefore, exploration of treatment alternatives is of clinical relevance. The present work focused on tumor cell-inhibiting effects of a combination of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and all trans retinoic acid (ATRA) in breast cancer cells.

Materials and methods: Breast cancer cell lines (BT-20, BT-474, MDA-MB-231, MDA-MB-436, MDA-MB-453, MCF-7, SKBR3, T47D, ZR-75-1) and the mammary epithelial cell line MCF-10A were treated with TRAIL and ATRA alone and in combination. Cell viability was assessed via 3-(4,5)-dimethylthiahiazo(-z-yl)-3,5-di-phenytetrazoliumromide (MTT) assay, the potential of cell colony formation via clonogenic assay, cell death induction via cell-cycle analysis by fluorescence-activated cell sorting (FACS), terminal deoxynucleotidyltransferase-mediated UTP nick end labeling (TUNEL) assay and Cell death detection ELISAPLUS, expression of apoptosis and TRAIL pathway proteins via western blot and cell surface expression of TRAIL receptor 1 (DR4) via FACS analysis.

Results: TRAIL and ATRA evoked synergistic inhibition of breast cancer cell viability based on cytostatic and cytotoxic mechanisms. This correlated with augmented fragmentation of nuclear DNA, up-regulation of TRAIL receptor, down-regulation of cyclin D1 and enhancement of caspase activity. MCF-10A cells were merely slightly susceptible to TRAIL and ATRA.

Conclusion: The cytostatic and cytotoxic effects of the combination of TRAIL and ATRA are tumor cell-selective.

Keywords: ATRA; TRAIL; apoptosis; breast cancer; cell death; retinoids; synergism.

MeSH terms

  • Apoptosis / drug effects
  • Breast Neoplasms / pathology*
  • Cell Cycle / drug effects
  • Cell Line, Tumor
  • DNA Fragmentation
  • Drug Resistance, Neoplasm
  • Drug Synergism
  • Epithelial Cells / drug effects
  • Female
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • In Situ Nick-End Labeling
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • Recombinant Proteins / pharmacology
  • TNF-Related Apoptosis-Inducing Ligand / pharmacology*
  • Tretinoin / pharmacology*
  • Tumor Stem Cell Assay

Substances

  • Neoplasm Proteins
  • Recombinant Proteins
  • TNF-Related Apoptosis-Inducing Ligand
  • TNFSF10 protein, human
  • Tretinoin