Kuhara Tomoe, Watanabe Daisuke, Akita Yoichi, Takeo Tomohiro, Ishida Natsuko, Nakano Aki, Yamashita Noriko, Ohshima Yuichiro, Kawada Morihiro, Yanagishita Takeshi, Tamada Yasuhiko, Matsumoto Yoshinari
Department of Dermatology, Aichi Medical University School of Medicine, Aichi, Japan.
Photodermatol Photoimmunol Photomed. 2008 Jun;24(3):142-6. doi: 10.1111/j.1600-0781.2008.00354.x.
BACKGROUND/PURPOSE: 5-aminolaevulinic acid-based photodynamic therapy (ALA-PDT) is widely performed in the clinical setting for superficial skin cancers, giving favorable results, but residual tumor and recurrence occur occasionally. Thioredoxin is a common antioxidant that suppresses apoptosis and facilitates cell growth. We investigated the expression of thioredoxin following ALA-PDT in human skin squamous cell carcinoma cell line, HSC-5.
ALA-PDT was performed in HSC-5 cells using low-dose (5 J/cm(2), 100 mW/cm(2)) or high-dose (30 J/cm(2), 100 mW/cm(2)) irradiation, and the expression of thioredoxin was measured by Western blotting. An MTT assay was used to assess cell growth following a low dose of multiple irradiations. Cell death was examined by Western blotting for caspase-3 and PARP. Immunofluorescence double staining using annexin V and propidium iodine was also performed.
Expression of thioredoxin was only observed following low-dose exposure ALA-PDT. Multiple low-dose exposure ALA-PDT significantly proliferated cell growth. With high-dose exposure ALA-PDT, caspase-3 and PARP expression were seen, and cell death due to apoptosis and/or necrosis was observed, but thioredoxin was barely detected.
Low-dose exposure ALA-PDT increased the expression of thioredoxin and facilitated the growth of HSC-5 cells.
背景/目的：基于5-氨基酮戊酸的光动力疗法（ALA-PDT）在浅表皮肤癌的临床治疗中广泛应用，效果良好，但偶尔会出现肿瘤残留和复发。硫氧还蛋白是一种常见的抗氧化剂，可抑制细胞凋亡并促进细胞生长。我们研究了ALA-PDT后硫氧还蛋白在人皮肤鳞状细胞癌细胞系HSC-5中的表达。
使用低剂量（5 J/cm²，100 mW/cm²）或高剂量（30 J/cm²，100 mW/cm²）照射对HSC-5细胞进行ALA-PDT，通过蛋白质免疫印迹法检测硫氧还蛋白的表达。采用MTT法评估低剂量多次照射后的细胞生长情况。通过蛋白质免疫印迹法检测caspase-3和PARP来检查细胞死亡情况。还进行了膜联蛋白V和碘化丙啶的免疫荧光双重染色。
仅在低剂量ALA-PDT照射后观察到硫氧还蛋白的表达。多次低剂量ALA-PDT照射显著促进了细胞生长。高剂量ALA-PDT照射后，可见caspase-3和PARP表达，观察到细胞因凋亡和/或坏死而死亡，但几乎检测不到硫氧还蛋白。
低剂量ALA-PDT照射增加了硫氧还蛋白的表达并促进了HSC-5细胞的生长。
