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.
Objective To study the effects of HMME-based photodynamic therapy on proliferation and apoptosis of rabbit
vascular smooth muscle cells(VSMCs). Method The cytotoxic effect of HMME-PDT on rabbit vascular smooth
muscle cells was studied by means of Trypan Blue assay, HMME at 10&mgr;g/ml concentration and the light dose at 2.4~4.8
J/cm2 were selected in the studies. The morphological character 24h post-PDT was investigated by HE Staining.
Annexin V and propidium iodide (PI) binding assays were performed to analyze the characteristics of cell death after
HMME-PDT. Furthermore, The intracellular distributions of the HMME were measured by the confocal laser scanning
microscope. Result It was showed the photocytotoxity to VSMC cells was dose related by Trypan Blue assay.
Histology observing suggests HMME-PDT could induce cell death through apoptosis or necrosis, and the apoptosic rate
was up to 50.5% by AnnexinV /PI assay. Moreover, the fluorescence images of HMME intracellular localization
demonstrated that the HMME diffused into the mitochondria. Conclusion HMME-PDT could significantly inhibite
VSMC proliferation and induce apoptosis.
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