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Cytotoxic Efficacy of Photodynamic Therapy in Osteosarcoma Cells In Vitro
Published on: March 18, 2014
Pyropheophorbide-α methyl ester-mediated photodynamic therapy triggers pyroptosis in osteosarcoma cells via the
Qiu Huang1, Yong Tao2, Ye Zhang2
1Department of Orthopaedics, The First Affiliated Hospital of Chongqing Medical University, Yuzhong, Chongqing, 400016, China; Chongqing Municipal Health Commission Key Laboratory of Musculoskeletal Regeneration and Translational Medicine, Yuzhong, Chongqing, 400016, China; Orthopaedic Research Laboratory of Chongqing Medical University, Yuzhong, Chongqing, 400016, China; Department of Orthopaedics, People's Hospital of Leshan, Leshan, Sichuan, 614000, China.
Background:
Pyropheophorbide-α methyl ester-mediated photodynamic therapy(MPPa-PDT) is a candidate treatment for solid tumors, including osteosarcoma. Pyroptosis has garnered significant attention in cancer research due to its pro-inflammatory and immunomodulatory nature. This study investigated the mechanism and role of MPPa-PDT-induced pyroptosis in osteosarcoma cells.
Methods:
We treated human osteosarcoma 143b and HOS cells with MPPa at concentrations of 0.5 μM and 0.25 μM, respectively, then irradiated the cells with LED light at 630 nm wavelength with an energy density of 4.8 J/cm2. Cell viability and apoptosis ratio were detected using CCK-8 and Annexin V-Propidium Iodide staining, respectively. Intracellular reactive oxygen species (ROS) levels and mitochondrial membrane potential (MtΔψ) were assessed using 2',7'-Dichlorofluorescin diacetate, and JC-1 staining kits, respectively. Scanning Electron Microscopy (SEM) was utilized to examine cell ultrastructure. The morphological changes of the cells were observed by an inverted microscope. Western blotting analysis was conducted to measure protein levels. To elucidate the mechanism and role, we re-evaluated relevant parameters after pretreating with NAC,Si caspase-3, and Si GSDME.
Results:
MPPa-PDT inhibited the activity of osteosarcoma 143b and HOS cells and induced pyroptosis with mitochondrial damage, ROS aggregation, and activation of Caspase-3 and GSDME. The effects of MPPa-PDT on the activity and apoptosis of osteosarcoma cells were partially reversed after pretreating with Si GSDME. After NAC pretreatment, the activation of pyroptosis and Caspase-3 induced by MPPa-PDT was partially reversed. After Si Caspase-3 pretreatment, the pyroptosis induced by MPPa-PDT was partially reversed.
Conclusion:
MPPa-PDT can induce pyroptosis in osteosarcoma cells, which has the effect of enhancing apoptotic processes. Mitochondrial damage and ROS/caspase-3/GSDME pathway are the possible mechanisms of pyroptosis induced by MPPa-PDT.
Insights
Pyropheophorbide-α methyl ester-mediated photodynamic therapy (MPPa-PDT) induces pyroptosis in osteosarcoma cells. This cell death pathway, involving mitochondrial damage and ROS/caspase-3/GSDME signaling, enhances apoptosis and offers a potential cancer treatment.
Area of Science:
- Oncology
- Biochemistry
- Cell Biology
Background:
- Pyropheophorbide-α methyl ester-mediated photodynamic therapy (MPPa-PDT) is explored for solid tumor treatment, including osteosarcoma.
- Pyroptosis, a pro-inflammatory cell death, is gaining attention for its immunomodulatory cancer research potential.
Purpose of the Study:
- To investigate the mechanism and role of MPPa-PDT-induced pyroptosis in osteosarcoma cells.
- To elucidate the signaling pathways involved in MPPa-PDT-induced cell death.
Main Methods:
- Osteosarcoma cells (143b, HOS) were treated with MPPa and irradiated.
- Assessed cell viability, apoptosis, reactive oxygen species (ROS), mitochondrial membrane potential (MtΔψ), and protein levels via Western blotting.
- Examined cellular ultrastructure using Scanning Electron Microscopy (SEM).
Main Results:
- MPPa-PDT inhibited osteosarcoma cell activity and induced pyroptosis, characterized by mitochondrial damage, ROS aggregation, and activation of Caspase-3 and GSDME.
- Pretreatment with N-acetylcysteine (NAC), Si caspase-3, or Si GSDME partially reversed MPPa-PDT-induced pyroptosis and cell death.
- MPPa-PDT induced pyroptosis, with effects partially reversed by Si GSDME and Si Caspase-3, suggesting their involvement.
Conclusions:
- MPPa-PDT effectively induces pyroptosis in osteosarcoma cells, augmenting apoptotic processes.
- Mitochondrial damage and the ROS/caspase-3/GSDME pathway are identified as key mechanisms driving MPPa-PDT-induced pyroptosis.
- This study highlights MPPa-PDT as a potential therapeutic strategy for osteosarcoma via pyroptosis induction.
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