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