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

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

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.

Citation Download Citation

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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KEYWORDS

Cell death

Photodynamic therapy

Flow cytometry

Breast cancer

Confocal microscopy

Laser scanners

Tissue optics

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