9-羟苯基叶啉-α 介导的光动力疗法通过 ROS 介导的 ERK 通路抑制诱导 Hep-2 细胞中基质金属蛋白酶-2（MMP-2）和 MMP-9 的下调。

9-Hydroxypheophorbide α-mediated photodynamic therapy induces matrix metalloproteinase-2 (MMP-2) and MMP-9 down-regulation in Hep-2 cells via ROS-mediated suppression of the ERK pathway.

机构信息

Department of Otorhinolaryngology-Head and Neck Surgery, Affiliated Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai, China.

Institute of Radiation Medicine, Fudan University, Shanghai, China.

出版信息

Photodiagnosis Photodyn Ther. 2014 Mar;11(1):55-62. doi: 10.1016/j.pdpdt.2013.12.001. Epub 2014 Jan 18.

DOI:10.1016/j.pdpdt.2013.12.001

PMID:24448387

Abstract

BACKGROUND

Photodynamic therapy (PDT) is a promising treatment modality for malignant diseases through the generation of reactive oxygen species (ROS). In this study, we assessed the change of migration and invasion of HEp-2 cells after sublethal doses of 9-hydroxypheophorbide α (9-HPbD)-mediated PDT in vitro, and explored the role of ROS in 9-HPbD-PDT-induced anti-metastatic effects in HEp-2 cells.

METHODS

Following PDT, ROS were measured by a fluorescence microscope in both the presence and absence of glutathione (GSH) pretreatment. Wound healing assay, cell migration assay, and matrigel invasion assay were used to evaluate the cellular migration and invasion. Western blot was performed to investigate the signaling pathways that may have been involved.

RESULTS

ROS were rapidly generated in 9-HPbD-loaded HEp-2 laryngeal cancer cells by the activation of a diode laser and were significantly inhibited by a 6-h GSH pretreatment. Wound healing assay, cell migration assay, and matrigel invasion assay showed that sublethal PDT significantly suppressed the migration and invasion of HEp-2 cells. GSH decreased the ability of PDT to inhibit the invasion of HEp-2 cells. Western blot analysis showed that PDT significantly inhibited the phosphorylation of MEK1/2 and ERK1/2, and significantly suppressed the expression of MMP-2 and MMP-9 after 24h following the implementation of sublethal PDT, and these efficacies of PDT could be abrogated by GSH pretreatment.

CONCLUSIONS

9-HPbD-PDT attenuated the migration and invasion of HEp-2 cells in vitro, which may be related to the down-regulated expression of MMP-2 and MMP-9 via ROS-mediated-inhibition of phosphorylation in the ERK/MEK signaling pathway.

摘要

背景

光动力疗法（PDT）通过产生活性氧（ROS）成为治疗恶性疾病的一种很有前途的治疗方式。在这项研究中，我们评估了亚致死剂量的 9-羟基 pheophorbide α（9-HPbD）介导的 PDT 后体外 HEp-2 细胞迁移和侵袭的变化，并探讨了 ROS 在 9-HPbD-PDT 诱导的 HEp-2 细胞抗转移效应中的作用。

方法

在存在和不存在谷胱甘肽（GSH）预处理的情况下，通过荧光显微镜测量 ROS。使用划痕愈合试验、细胞迁移试验和基质胶侵袭试验评估细胞迁移和侵袭。通过 Western blot 分析研究可能涉及的信号通路。

结果

二极管激光激活后，9-HPbD 负载的喉癌细胞中迅速产生 ROS，并且 6 小时 GSH 预处理可显著抑制 ROS 的产生。划痕愈合试验、细胞迁移试验和基质胶侵袭试验表明，亚致死 PDT 显著抑制了 HEp-2 细胞的迁移和侵袭。GSH 降低了 PDT 抑制 HEp-2 细胞侵袭的能力。Western blot 分析显示，亚致死 PDT 后 24 小时，PDT 显著抑制了 MEK1/2 和 ERK1/2 的磷酸化，并显著抑制了 MMP-2 和 MMP-9 的表达，而 GSH 预处理可以消除 PDT 的这些功效。