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<p>Three-dimensional (3D) cell culture models are increasingly used in biomedicine, while two-dimensional (2D) systems represent a simplified approach with limited intercellular interactions and no spatial organization. The aim of this work was to compare the response of tumor cells to cytotoxic chemical complexes in 2D and 3D conditions, with a special focus on apoptosis. The results showed a pronounced paradox: although proapoptotic markers (Bax/Bcl-2 ratio and caspase-3) were higher in the 2D system, the phenotypic outcome indicated a higher frequency of late apoptosis in 3D conditions, with a lower percentage of viable cells. These findings indicate that molecular markers do not directly reflect the outcome of cell death, but the apoptotic response depends on the spatial context. The mathematical model offered additional quantitative insight that may help to contextualize these differences, suggesting localized hypoxic and metabolic adaptation in 3D systems. In conclusion, in HCT-116 treated with Au(III) complexes, 2D and 3D models represent qualitatively different modes of response. These observations support the hypothesis that apoptosis execution is shaped by spatial context, pending validation in additional cell lines and platforms.</p>

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Apoptosis execution is spatially regulated

  • 期刊论文

  • 发表日期：

    2026年6月18日

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  • Nevena Milivojević Dimitrijević,
  • Ana Mirić,
  • Tijana Đukić,
  • Ivica Petrović,
  • Ana Kesić,
  • Vladimir Jurišić,
  • Luka Dragačević,
  • Marko N. Živanović,
  • Nenad Filipović

摘要

Three-dimensional (3D) cell culture models are increasingly used in biomedicine, while two-dimensional (2D) systems represent a simplified approach with limited intercellular interactions and no spatial organization. The aim of this work was to compare the response of tumor cells to cytotoxic chemical complexes in 2D and 3D conditions, with a special focus on apoptosis. The results showed a pronounced paradox: although proapoptotic markers (Bax/Bcl-2 ratio and caspase-3) were higher in the 2D system, the phenotypic outcome indicated a higher frequency of late apoptosis in 3D conditions, with a lower percentage of viable cells. These findings indicate that molecular markers do not directly reflect the outcome of cell death, but the apoptotic response depends on the spatial context. The mathematical model offered additional quantitative insight that may help to contextualize these differences, suggesting localized hypoxic and metabolic adaptation in 3D systems. In conclusion, in HCT-116 treated with Au(III) complexes, 2D and 3D models represent qualitatively different modes of response. These observations support the hypothesis that apoptosis execution is shaped by spatial context, pending validation in additional cell lines and platforms.