The photodamage effect and ROS generation induced by PDT with HMME in MCF-7cells in vitro

Huijuan Yin; Xiaoyuan Li; Jianzhong Liu; Yan Li

doi:10.1117/12.741599

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1 May 2007 The photodamage effect and ROS generation induced by PDT with HMME in MCF-7cells in vitro

Huijuan Yin, Xiaoyuan Li, Jianzhong Liu, Yan Li

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Proceedings Volume 6534, Fifth International Conference on Photonics and Imaging in Biology and Medicine; 65341Z (2007) https://doi.org/10.1117/12.741599
Event: Fifth International Conference on Photonics and Imaging in Biology and Medicine, 2006, Wuhan, China

Abstract

Hematoporphyrin monomethyl ether (HMME) is a novel and promising porphyrin-related photosensitizer for photodynamic therapy (PDT). We use the human breast cancer MCF-7 cells to investigate the photodamage effect of HMME and reactive oxygen species (ROS) generation in HMME-PDT. Methods: The growth rates of MCF-7 cells at 24h after irradiation by 532nm laser with HMME of 5~20μg/ml and light dose of 0.3~4.8J/cm² were determined by CCK-8 assays. Hoechst33342 staining was used to investigate the morphological change of the tumor cell. Flow cytometry combined with dual Annexin V/PI staining was used to identify the death mode of the cells following PDT. The changes of ROS labeled by DCFH-DA were observed by Laser Scanning Confocal Microscopy (LSCM). Our results show that HMME-based PDT induced significant cell death, and the photocytotoxity to MCF-7 cells is dose-dependent at the range of HMME concentration 5~20μg/ml and the light dose 0.3~4.8J/cm². The nucleolus underwent apoptosis and/or necrosis observed by LSCM with Hoechst33342 staining. The necrosis and apoptosis rate were 16.0% and 12.4% respectively by FCM, showing the number of necrosic cells was more than that of apoptosis. There was an intense increase of fluorescence intensity standing for ROS generation within 30min post-PDT, and the peak was at about 10min after PDT. Our results suggest that HMME-PDT could inhibit the proliferation of MCF-7 cells remarkably. Because the MCF-7 cells lack procaspase-3, the apoptosis rate is lower. ROS played an important role in the photodamage with HMME.

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Huijuan Yin, Xiaoyuan Li, Jianzhong Liu, and Yan Li "The photodamage effect and ROS generation induced by PDT with HMME in MCF-7cells in vitro", Proc. SPIE 6534, Fifth International Conference on Photonics and Imaging in Biology and Medicine, 65341Z (1 May 2007); https://doi.org/10.1117/12.741599

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