Isik Seyma, Ozcesmeci Mukaddes, Burat Ayfer Kalkan, Hamuryudan Esin, Erdogmus Ali, Can Ozge, Serhatli Muge
Department of Medical Biotechnology, Graduate School of Health Sciences, Acibadem Mehmet Ali Aydinlar University, 34752, Istanbul, Turkey.
TUBITAK Marmara Research Center, Climate Change and Life Sciences, Biotechnology Research Group, 41470, Kocaeli, Turkey.
Sci Rep. 2025 Jul 11;15(1):25148. doi: 10.1038/s41598-025-09630-7.
In this study, the PDT activities of 2,9(10),16(17),23(24)-tetrakis(2-dimethylammoniumethoxy) phthalocyaninatozinc(II) tetraiodide (ZnPc1) and 1,8(11),15(18),22(25)-tetrakis(2-trimethylaminoethoxy)phthalocyaninatozinc(II)tetraiodide (ZnPc2) compounds were investigated in 2D monolayer cultures and 3D spheroids of three human cancer cell lines: human submaxillary salivary gland epidermoid carcinoma (A253), human colon colorectal adenocarcinoma (HT29), and human pharynx squamous carcinoma (FaDu) cells. The results indicate that both molecules are non-toxic in the absence of light, which is a crucial feature of an effective photosensitizer. Upon exposure to light, ZnPc1 exhibited significant cytotoxicity in all three cell lines, particularly in FaDu cells, in both 2D monolayer cultures and 3D spheroids, whereas ZnPc2 showed moderate efficacy compared to ZnPc1. PDT using both phthalocyanine (Pc) molecules resulted in substantial reactive oxygen species (ROS) production. Delayed ROS production is higher than that of immediate ROS, indicating their ability to stimulate ROS production over an extended period and retain an oxidative stress response in the cells rather than immediately after PDT. Among these molecules, ZnPc1 induced both immediate and delayed ROS production more efficiently than ZnPc2. Furthermore, singlet oxygen yields of ZnPc1 were higher than ZnPc2, which is consistent with the cytotoxicity results. These findings confirmed that PDT induces an ROS-mediated cytotoxic response. The mechanisms of cellular death triggered by PDT were evaluated, and the results revealed that apoptosis was the predominant process. These findings underscore the potential of ZnPc1 as a potent photosensitizer in PDT while also highlighting the differences between 2D and 3D culture models in evaluating PDT efficacy. While the 2D system enables simplified cytotoxicity evaluation, the 3D spheroid model better replicates physiologically relevant environment and treatment resistance. This comparison underscores necessity of integrating 3D models in PDT studies for more predictive in vivo insights.
在本研究中，对2,9(10),16(17),23(24)-四(2-二甲基铵乙氧基)酞菁锌(II)四碘化物(ZnPc1)和1,8(11),15(18),22(25)-四(2-三甲基氨基乙氧基)酞菁锌(II)四碘化物(ZnPc2)化合物在三种人类癌细胞系的二维单层培养物和三维球体中的光动力疗法（PDT）活性进行了研究，这三种癌细胞系分别是人下颌下唾液腺表皮样癌(A253)、人结肠直肠腺癌(HT29)和人咽鳞状癌(FaDu)细胞。结果表明，在无光条件下这两种分子均无毒性，这是有效光敏剂的关键特性。在光照下，ZnPc1在二维单层培养物和三维球体中对所有三种细胞系均表现出显著的细胞毒性，尤其是在FaDu细胞中，而与ZnPc1相比，ZnPc2显示出中等疗效。使用这两种酞菁(Pc)分子进行光动力疗法均导致大量活性氧(ROS)生成。延迟ROS生成高于即时ROS生成，表明它们能够在较长时间内刺激ROS生成并在细胞中保持氧化应激反应，而不是在光动力疗法后立即产生。在这些分子中，ZnPc1比ZnPc2更有效地诱导即时和延迟ROS生成。此外，ZnPc1的单线态氧产率高于ZnPc2，这与细胞毒性结果一致。这些发现证实了光动力疗法诱导ROS介导的细胞毒性反应。对光动力疗法引发的细胞死亡机制进行了评估，结果显示凋亡是主要过程。这些发现强调了ZnPc1作为光动力疗法中一种有效的光敏剂的潜力，同时也突出了二维和三维培养模型在评估光动力疗法疗效方面的差异。虽然二维系统能够简化细胞毒性评估，但三维球体模型能更好地模拟生理相关环境和治疗抗性。这种比较强调了在光动力疗法研究中整合三维模型以获得更具预测性的体内见解的必要性。
