| Objective:Photodynamic therapy (PDT) is a promising treatment modality for malignant disease through the generation of reactive oxygen species (ROS). To assess the change of migration and invasion about the HEp-2cells after sublethal doses of9-hydroxypheophorbide a (9-HPbD)-mediated PDT in vitro, and to explore the role of ROS in9-HPbD-PDT-induced anti-metastatic effects in HEp-2cells.Methods:Following PDT, ROS were measured by a fluorescence microscope with or without pretreatment of GSH. Wound healing assay, cell migration assay, and matrigel invasion assay were respectively used to evaluate the cellular migration and invasion. Western blot was performed to investigate the signaling pathways that may have been involved.Results:Diode laser (664nm with an energy density of2.0J/cm2) was used to activate9-HPbD-incubated Hep-2laryngeal cancer cells. Following PDT, ROS were rapidly generated in9-HPbD-loaded HEp-2laryngeal cancer cells by the activation of diode laser and were significantly inhibited by a6-h pretreatment of GSH. Wound healing assay, cell migration assay, and matrigel invasion assay showed that sublethal PDT significantly suppressed the migration and invasion of HEp-2cells. GSH decreased the ability of PDT to inhibit the invasion of HEp-2cells. Western blot analysis showed that PDT significantly inhibited the phosphorylation of MEK1/2and ERK1/2, and significantly suppressed the expression of MMP-2and MMP-9after24hours 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-2cells which may relate to the down-regulated expression of MMP-2and MMP-9via ROS-mediated-inhibition of phosphorylation in the ERK/MEK signaling pathway. |
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