Department of Dermatology, Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
Department of Dermatology, Forest Industry Worker Hospital of Shaanxi Province, Xi'an, China.
Braz J Med Biol Res. 2020;53(5):e8457. doi: 10.1590/1414-431x20208457. Epub 2020 Apr 27.
The objective of this study was to investigate whether the conjugation of gold nanoparticles (GNPs) to 5-aminolevulinic acid (5-ALA) could enhance the anti-tumor efficiency of photodynamic therapy (PDT) in epidermoid carcinoma cells. The mRNA and protein expression levels were determined by quantitative real-time PCR and western blot, respectively. Cell viability, apoptosis, invasion, and migration were determined by MTT assay, flow cytometry, transwell invasion assay, and migration assay, respectively. Singlet oxygen generation was detected by the singlet oxygen sensor green reagent assay. Our results showed that PDT with 5-ALA and GNPs-conjugated 5-ALA (5-ALA-GNPs) significantly suppressed cell viability, increased cell apoptosis and singlet oxygen generation in both HaCat and A431 cells, and PDT with 5-ALA and 5-ALA-GNPs had more profound effects in A431 cells than that in HaCat cells. More importantly, 5-ALA-GNPs treatment potentiated the effects of PDT on cell viability, cell apoptosis, and singlet oxygen generation in A431 cells compared to 5-ALA treatment. Further in vitro assays showed that PDT with 5-ALA-GNPs significantly decreased expression of STAT3 and Bcl-2 and increased expression of Bax in A431 cells compared with PDT with 5-ALA. In addition, 5-ALA-GNPs treatment enhanced the inhibitory effects of PDT on cell invasion and migration and Wnt/β-catenin signaling activities in A431 cells compared to 5-ALA treatment. In conclusion, our results suggested that GNPs conjugated to 5-ALA significantly enhanced the anti-tumor efficacy of PDT in A431 cells, which may represent a better strategy to improve the outcomes of patients with cutaneous squamous cell carcinoma.
本研究旨在探讨金纳米粒子（GNPs）与 5-氨基酮戊酸（5-ALA）缀合是否可以增强光动力疗法（PDT）在表皮癌细胞中的抗肿瘤效率。通过定量实时 PCR 和 Western blot 分别测定 mRNA 和蛋白质表达水平。通过 MTT 测定法、流式细胞术、Transwell 侵袭测定法和迁移测定法分别测定细胞活力、细胞凋亡、侵袭和迁移。通过单线态氧传感器绿试剂测定法检测单线态氧的产生。我们的结果表明，5-ALA 和 GNPs 缀合的 5-ALA（5-ALA-GNPs）的 PDT 显著抑制了 HaCat 和 A431 细胞中的细胞活力，增加了细胞凋亡和单线态氧的产生，并且 5-ALA 和 5-ALA-GNPs 的 PDT 在 A431 细胞中的作用比 HaCat 细胞中的作用更为显著。更重要的是，与 5-ALA 处理相比，5-ALA-GNPs 处理增强了 A431 细胞中 PDT 对细胞活力、细胞凋亡和单线态氧产生的影响。进一步的体外实验表明，与 5-ALA 的 PDT 相比，5-ALA-GNPs 的 PDT 显著降低了 A431 细胞中 STAT3 和 Bcl-2 的表达，增加了 Bax 的表达。此外，与 5-ALA 处理相比，5-ALA-GNPs 处理增强了 PDT 对 A431 细胞侵袭和迁移以及 Wnt/β-catenin 信号活性的抑制作用。总之，我们的结果表明，与 5-ALA 缀合的 GNPs 显著增强了 A431 细胞中 PDT 的抗肿瘤疗效，这可能代表了改善皮肤鳞状细胞癌患者预后的更好策略。
