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Cell Line-Dependent Cell Death Pathways Induced by Thymoquinone in Colorectal Cancer Cells.

Natalia Kurowska1, Maria Książek1, Paulina Borkowska2

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Thymoquinone (TQ) reduces colorectal cancer (CRC) cell viability through distinct cell death pathways, depending on the cancer

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5-fluorouracilapoptosiscaspase-independent cell deathcell death pathwayscolorectal cancermolecular mechanismsnecrosisthymoquinone

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Area of Science:

  • Oncology
  • Molecular Biology
  • Pharmacology

Background:

  • Colorectal cancer (CRC) is a major cause of cancer mortality.
  • Resistance to 5-fluorouracil (5-FU) is a significant therapeutic hurdle in CRC treatment.
  • Thymoquinone (TQ), derived from Nigella sativa, shows anticancer potential, but its cell death mechanisms are context-dependent.

Purpose of the Study:

  • To investigate the cell line-specific death pathways induced by TQ in colorectal cancer models.
  • To compare TQ's effects in 5-FU-sensitive (RKO) and 5-FU-resistant (SW1116) CRC cell lines.
  • To assess TQ's impact on normal colon epithelial cells.

Main Methods:

  • Cell viability assays (MTT)
  • DNA fragmentation analysis
  • Caspase activity assays (caspase-3/7, -8, -9)
  • Cell death phenotype analysis (flow cytometry)
  • Gene expression analysis (RT-qPCR) for apoptosis and necroptosis markers

Main Results:

  • TQ reduced viability in both RKO and SW1116 cells with minimal toxicity to normal cells.
  • In RKO cells (microsatellite instability), TQ induced apoptosis via caspase activation and pro-apoptotic gene upregulation.
  • In SW1116 cells (chromosomal instability), TQ triggered caspase-independent necrotic cell death.
  • Combined TQ and 5-FU treatment showed no synergistic cytotoxicity but distinct cell death programs.

Conclusions:

  • TQ induces cancer cell death through context-dependent mechanisms, primarily apoptosis in MSI cells and necrosis in CIN cells.
  • TQ's efficacy is modulated by the molecular background of colorectal cancer cells.
  • TQ does not enhance 5-FU efficacy synergistically but activates distinct cell death pathways.