School of Medical and Health Sciences, Tung Wah College, Hong Kong, China.
School of Medical and Health Sciences, Tung Wah College, Hong Kong, China.
J Photochem Photobiol B. 2020 Sep;210:111987. doi: 10.1016/j.jphotobiol.2020.111987. Epub 2020 Aug 7.
Photodynamic Therapy (PDT) offers an alternative option for the treatment of nasopharyngeal carcinoma (NPC). The utilization of 3-dimensional (3D) culture model might provide better understanding of PDT effects on NPC cells. The aim of this in vitro study was to compare PDT effect on NPC cells using 2D and 3D models. Two 3D culture models were established using liquid overlay method with agarose base (MCL) and hanging drop method (MCS). PDT was carried out using the combination of FosPeg® and 652 nm laser in 3D and conventional 2D models. The effects of 3D culture size and morphology on the uptake and distribution of sensitizer and gene expression were examined. Photocytotoxity, mode of cell death, and protein expression were compared for 2D and 3D models. Regular and irregular NPC spheroids were obtained from MCL and MCS methods, respectively. A significantly down-regulation of LMP1 mRNA were observed in MCL spheroid. The sensitizer uptake in 3D spheroids was half of 2D culture. More sensitizers were required to obtain IC in 3D spheroids. Apoptosis, necrosis and autophagosomes were detected in PDT treated 2D and 3D cells. Different protein expression patterns were observed in 2D and 3D models. FosPeg® PDT is effective in killing NPC cells. MCL-derived 3D spheroid model is more suitable for the evaluation of PDT killing mechanisms.
光动力疗法(PDT)为鼻咽癌(NPC)的治疗提供了另一种选择。使用三维(3D)培养模型可能有助于更好地了解 PDT 对 NPC 细胞的影响。本体外研究的目的是比较使用 2D 和 3D 模型对 NPC 细胞的 PDT 效果。使用琼脂糖基底液覆盖法(MCL)和悬滴法(MCS)建立了两种 3D 培养模型。在 3D 和常规 2D 模型中使用 FosPeg®和 652nm 激光组合进行 PDT。检查了 3D 培养大小和形态对敏化剂摄取和分布以及基因表达的影响。比较了 2D 和 3D 模型的光细胞毒性、细胞死亡方式和蛋白质表达。从 MCL 和 MCS 方法中分别获得了规则和不规则的 NPC 球体。在 MCL 球体中观察到 LMP1 mRNA 的显著下调。3D 球体中的敏化剂摄取量是 2D 培养的一半。需要更多的敏化剂才能在 3D 球体中获得 IC。在 PDT 处理的 2D 和 3D 细胞中检测到凋亡、坏死和自噬体。在 2D 和 3D 模型中观察到不同的蛋白质表达模式。FosPeg®PDT 可有效杀死 NPC 细胞。MCL 衍生的 3D 球体模型更适合评估 PDT 杀伤机制。