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  • Molecular and Cellular Evaluation of Apoptosis and Tight Junction Proteins (CLDN5/ZO1) in a Glioblastoma-BBB Co-culture Model Following Sensitizer-Free Phototherapy

  • Mohammad Reza Rashidian Vaziri,1 Armin Imanparast,2 Ameneh Sazgarnia,3 Samaneh Sharif,4 Andrey Terskov,5,* Alexander Shirokov,6
    1. Department of Physics, Faculty of Sciences, Ferdowsi University of Mashhad, Mashhad, Iran
    2. Medical Physics Research Center, Basic Sciences Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran & Department of Medical Physics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
    3. Medical Physics Research Center, Basic Sciences Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran & Department of Medical Physics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
    4. Medical Genetics Research Center, Basic Sciences Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran & Department of Medical Genetics, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
    5. Department of Biology, Saratov State University, Astrakhanskaya Str. 83, 410012 Saratov, Russia
    6. Department of Biology, Saratov State University, Astrakhanskaya Str. 83, 410012 Saratov, Russia


  • Introduction: Glioblastoma (GB) is an aggressive brain tumor requiring novel therapeutic approaches. While photodynamic therapy is promising, distinguishing viable tumor cells and utilizing exogenous photosensitizers present clinical challenges. This study evaluates the molecular and cellular responses of GB cells to a sensitizer-free phototherapy targeting endogenous oxygen.
  • Methods: A co-culture model comprising C6 glioma cells and an in vitro blood-brain barrier (BBB) was utilized. The models were exposed to 1267 nm near-infrared irradiation. We assessed intracellular reactive oxygen species (ROS) production, the expression of tight junction (TJ) proteins (CLDN5 and ZO1) via immunohistochemistry, and apoptosis rate using the TUNEL assay.
  • Results: The targeted irradiation significantly enhanced ROS generation, revealing a 2.0-fold increase in the BBB-C6 glioma cells. Molecular evaluation demonstrated a significant downregulation in the expression of TJ proteins, CLDN5 and ZO1, in the BBB-C6 model compared to non-irradiated controls. Furthermore, the treatment induced a significant cellular response, triggering apoptosis in $27pm4\%$ of the glioma cells compared to $16pm3\%$ in the control group, while markedly suppressing tumor cell proliferation.
  • Conclusion: Sensitizer-free photoactivation effectively modulates the expression of essential barrier proteins (CLDN5/ZO1) and selectively induces apoptosis in glioma cells. These molecular alterations highlight a potential novel adjuvant strategy for glioblastoma management.
  • Keywords: Glioblastoma, Blood-Brain Barrier, Sensitizer-Free Phototherapy, Apoptosis, Tight Junction Proteins

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