Analysis of apoptotic effects induced by photodynamic therapy in a human biliary cancer cell line.
Analysis of apoptotic effects induced by photodynamic therapy in a human biliary cancer cell line.
复制标题
DOI：
--
复制
发表时间：
2010-06
影响因子：
2
通讯作者：
T. Nonaka;A. Nanashima;M. Nonaka;M. Uehara;H. Isomoto;I. Asahina;T. Nagayasu
中科院分区：
文献类型：
--
作者：
T. Nonaka;A. Nanashima;M. Nonaka;M. Uehara;H. Isomoto;I. Asahina;T. Nagayasu
背景光动力疗法（PDT）是一种治疗胆道癌的相对较新的方法，迄今为止尚未对其效果进行详细研究。本研究旨在探讨PDT诱导人胆管癌细胞凋亡的机制。材料和方法在体外，NOZ细胞与porfimer钠（Photofrin）孵育长达24小时，然后暴露于激光。PDT后使用甲基四唑测定法评估细胞活力。采用DNA片段化、细胞周期分析和caspase-3活性测定等方法检测PDT诱导的细胞凋亡。在体内，用TUNEL法检测DNA片段化。结果24 h内可见DNA梯状条带形成，caspase-3活性增强。PDT后24小时，流式细胞仪分析DNA断裂的细胞比例显著增加至22.2%。在体内模型中，TUNEL阳性细胞从PDT后6小时开始在植入的肿瘤中增加，并在12小时后达到峰值。结论Photofrin光动力疗法对人胆管癌细胞株具有抗肿瘤作用，并诱导细胞凋亡。
BACKGROUND Photodynamic therapy (PDT) is a relatively new approach for the treatment of biliary tract carcinoma, and its effects have not been investigated in detail to date. This study investigated the mechanisms of human biliary cancer cell death by PDT by focusing on apoptosis induction in vitro and in vivo. MATERIALS AND METHODS In vitro, NOZ cells were incubated with porfimer sodium (Photofrin) for up to 24 hours before exposure to laser light. Cell viability was assessed using a methyltetrazolium assay after PDT. DNA fragmentation, cell cycle analysis and caspase-3 activity assay were performed to evaluate apoptotic cells induced by PDT. In vivo, DNA fragmentation was detected by TUNEL assay. RESULTS DNA ladder formation and activation of caspase-3 were observed within 24 hours. The proportion of cells with DNA fragmentation on flow cytometric analysis was increased significantly to 22.2% at 24 hours after PDT. In the in vivo model, TUNEL-positive cells began to increase in the implanted tumour from 6 hours after PDT, and peaked 12 hours later. CONCLUSION PDT with Photofrin in this human biliary cancer cell line has antitumor effects and induces apoptotic cell death after PDT.