Sato Taiki, Sato Kazuomi
Graduate School of Agriculture, Tamagawa University, 6-1-1 Machida, Tokyo 194-8610, Japan.
Biosystems & Biofunctions Research Center, Tamagawa University Research Institute, 6-1-1 Machida, Tokyo 194-8610, Japan.
Biology (Basel). 2022 Nov 30;11(12):1743. doi: 10.3390/biology11121743.
Several studies have suggested the potential benefits of 5-aminolevulinic acid (5-ALA)-based photodynamic therapy (PDT). 5-ALA is a precursor of heme, which generates reactive oxygen species (ROS) following photoirradiation. Some reports indicate that blue light induces intracellular ROS production. In the present study, we elucidated the effects of blue light and 5-ALA on DNA integrity in B16F1 murine melanoma and human keratinocyte HaCaT cells using a variety of comet assay techniques. Co-treatment with blue light and 5-ALA significantly decreased cell viability in both cell lines. A neutral comet assay was performed to assess DNA double-strand break (DSB) formation and blue light and 5-ALA caused DSBs. We also performed an alkali comet assay to detect single-strand breaks (SSB) and alkali labile sites (ALS). The results indicated that 5-ALA accelerated blue light-induced SSB formation. In addition, modified comet assays were done using two types of enzymes to evaluate oxidative DNA damages. The results indicated that blue light and 5-ALA generated oxidized purine and pyrimidines in both cell lines. In summary, co-treatment with 5-ALA and photoirradiation may cause unexpected DNA damage in cells and tissues.
多项研究表明了基于5-氨基乙酰丙酸(5-ALA)的光动力疗法(PDT)的潜在益处。5-ALA是血红素的前体,在光照射后会产生活性氧(ROS)。一些报告表明蓝光可诱导细胞内ROS生成。在本研究中,我们使用多种彗星试验技术阐明了蓝光和5-ALA对B16F1小鼠黑色素瘤细胞和人角质形成细胞HaCaT细胞中DNA完整性的影响。蓝光和5-ALA联合处理显著降低了两种细胞系的细胞活力。进行中性彗星试验以评估DNA双链断裂(DSB)的形成,蓝光和5-ALA导致了DSB的产生。我们还进行了碱性彗星试验以检测单链断裂(SSB)和碱不稳定位点(ALS)。结果表明5-ALA加速了蓝光诱导的SSB形成。此外,使用两种酶进行改良彗星试验以评估氧化性DNA损伤。结果表明蓝光和5-ALA在两种细胞系中均产生了氧化的嘌呤和嘧啶。总之,5-ALA与光照射联合处理可能会在细胞和组织中造成意想不到的DNA损伤。