de Almeida Rainara Moreno Sanches, Fontana Letícia Corrêa, Dos Santos Vitorio Gabrielle, Pereira André Henrique Correia, Soares Cristina Pacheco, Pinto Juliana Guerra, Ferreira-Strixino Juliana
Laboratory of Photobiology Applied to Health, Institute of Research and Development, University of Vale do Paraíba, Av. Shishima Hifumi, 2911, Urbanova, CEP: 12244-000, São José dos Campos, SP, Brazil.
Laboratory of Cell Compartment Dynamics, Institute of Research and Development, University of Vale do Paraíba, Av. Shishima Hifumi, 2911, Urbanova, CEP: 12244-000, São José dos Campos, SP, Brazil.
Photodiagnosis Photodyn Ther. 2020 Jun;30:101685. doi: 10.1016/j.pdpdt.2020.101685. Epub 2020 Feb 9.
Gliosarcoma is a highly aggressive malignant neoplasm and a histopathological variant of wild-type glioblastoma multiforme isocitrate dehydrogenase (HDI). The current standard treatment consists of chemotherapy, radiotherapy and surgical resection, however, despite advances in these techniques, the patient's prognosis remains unfavorable. Photodynamic therapy (PDT) is a noninvasive technique that has been highlighted as an alternative form of cancer treatment because it does not present the side effects associated with systemic treatments. The objective of this study was to evaluate the cell viability and the intracellular localization of photosensitizer (PS) chlorin e6 Fotoenticine in 9L/lacZ cells. Therefore, tests of cytotoxicity, morphology, and location of PS were performed. The viability test showed no cytotoxicity in the dark at all concentrations and 100 % cell death at the highest concentrations after PDT. The mitochondrial activity test showed a reduction in all groups after PDT. The production of reactive oxygen species (ROS) was higher in the PDT groups and dependent on the PS concentration. Morphological analysis after PDT showed apparent cytoplasmic destruction in all the tested concentrations, with the presence of rounded cells due to the loss of their extensions and absence of nuclear alterations. The PS accumulation in the mitochondria and cytoskeleton was observed by the confocal microscopy; however, there is no evidence of its internalization in the lysosomes. It was concluded that PDT with Fotoenticine is a promising alternative therapy showing decreased cell viability, increased ROS production and adequate localization to trigger cell death.
胶质肉瘤是一种高度侵袭性的恶性肿瘤，是野生型多形性胶质母细胞瘤异柠檬酸脱氢酶（HDI）的一种组织病理学变体。目前的标准治疗包括化疗、放疗和手术切除，然而，尽管这些技术取得了进展，但患者的预后仍然不佳。光动力疗法（PDT）是一种非侵入性技术，因其不具有与全身治疗相关的副作用而被视为一种替代癌症治疗方法。本研究的目的是评估光敏剂（PS）二氢卟吩e6（Fotoenticine）在9L/lacZ细胞中的细胞活力和细胞内定位。因此，进行了细胞毒性、形态学和PS定位测试。活力测试表明，在黑暗中所有浓度下均无细胞毒性，而在PDT后最高浓度下细胞死亡率为100%。线粒体活性测试表明，PDT后所有组的活性均降低。PDT组中活性氧（ROS）的产生更高，且依赖于PS浓度。PDT后的形态学分析表明，在所有测试浓度下均出现明显的细胞质破坏，由于细胞突起丧失而出现圆形细胞，且无核改变。通过共聚焦显微镜观察到PS在线粒体和细胞骨架中的积累；然而，没有证据表明其被溶酶体内化。得出的结论是，Fotoenticine介导的PDT是一种有前景的替代疗法，可降低细胞活力、增加ROS产生并实现触发细胞死亡的充分定位。
