Tirapelli Luis Fernando, Morgueti Marcelo, da Cunha Tirapelli Daniela Pretti, Bagnato Vanderlei Salvador, Ferreira Juliana, Neto Fermino Sanches Lizarte, Peria Fernanda Maris, Oliveira Harley Francisco, Junior Carlos Gilberto Carlotti
Department of Surgery and Anatomy, University of São Paulo (USP), Brazil.
Photomed Laser Surg. 2011 May;29(5):305-9. doi: 10.1089/pho.2009.2649. Epub 2011 Jan 23.
Despite significant advances in neurosurgical techniques, the median survival time of patients with glioblastoma has improved little over the past 50 years and remains less than one year. Photodynamic therapy (PDT) is presently established as a widely accepted modality for the treatment of a variety of solid tumors.
This study evaluated the effect of PDT-Photogem(®) on five glioma cell lines (U87, U138, U251, U343, and T98G).
The experiments were carried out in 25-cm(3) flasks with different groups of cells seeded at a density of 1 × 10(5) cells per flask. After 3 h, the medium was removed, and the cells were incubated for 4 h with Photogem (5 μg/mL). After the incubation time, the photosensitizer-containing medium was removed and the cells were irradiated with LED (630 nm, 25 mW/cm(2), 25 J/cm(2)) devices for 17 min. For the final steps of the PDT, the cells were returned to the incubator and kept at 37°C with 5% CO(2) for 24 h, the cell viability assay was assessed using the trypan blue method, and the expression of Caspase 3 mRNA levels was assessed by real-time quantitative PCR.
Upon PDT-Photogem(®) treatment, viable cells, as evaluated by the trypan blue dye-exclusion method, decreased in two cell lines (U87 and U138) but not in the other three. Apoptosis, as assessed by the expression of caspase-3 mRNA levels, was at least partly involved in the death mechanism of the cell lines.
Collectively, our results indicated that PDT-Photogem(®) can act in glioma cells, thus encouraging new experiments in this field.
尽管神经外科技术取得了显著进展，但胶质母细胞瘤患者的中位生存时间在过去50年里几乎没有改善，仍不足一年。光动力疗法（PDT）目前已成为治疗多种实体瘤的广泛接受的方式。
本研究评估了PDT-Photogem（®）对五种胶质瘤细胞系（U87、U138、U251、U343和T98G）的作用。
实验在25 cm³的培养瓶中进行，不同组的细胞以每瓶1×10⁵个细胞的密度接种。3小时后，去除培养基，将细胞与Photogem（5 μg/mL）孵育4小时。孵育时间结束后，去除含光敏剂的培养基，用LED（630 nm，25 mW/cm²，25 J/cm²）设备对细胞照射17分钟。在PDT的最后步骤中，将细胞放回培养箱，在37°C和5% CO₂条件下培养24小时，使用台盼蓝法评估细胞活力测定，并通过实时定量PCR评估Caspase 3 mRNA水平的表达。
经PDT-Photogem（®）处理后，通过台盼蓝染料排除法评估，两个细胞系（U87和U138）中的活细胞减少，但其他三个细胞系中没有减少。通过caspase-3 mRNA水平的表达评估的细胞凋亡至少部分参与了细胞系的死亡机制。
总体而言，我们的结果表明PDT-Photogem（®）可作用于胶质瘤细胞，从而鼓励在该领域进行新的实验。
