研究目的
To compare the susceptibility of human melanoma cells to photoactivated Hypericin (Hyp) and aluminium(III) phthalocyanine chloride tetrasulphonate (AlPcS4Cl) in terms of cellular toxicity, subcellular localization, and photodynamic efficacy.
研究成果
Hyp-PDT is more effective than AlPcS4Cl-PDT in inducing apoptosis in melanoma cells. The efficiency of PDT depends on the PS concentration, incubation time, and light dose.
研究不足
The study is limited to in vitro conditions and may not fully replicate the complexity of in vivo environments. The focus was on a single cell line (A375), and results may vary with other melanoma cell lines.
1:Experimental Design and Method Selection:
The study involved treating melanoma cells (A375 cell line) with various concentrations of Hyp or AlPcS4Cl alone, laser irradiation alone, or a combination of both. Changes in cell morphology, viability, membrane integrity, and proliferation were assessed.
2:Sample Selection and Data Sources:
Human malignant melanoma (A375 cell line) was used. Cells were cultured in supplemented DMEM.
3:List of Experimental Equipment and Materials:
Photosensitizers (Hyp and AlPcS4Cl), diode lasers (594 nm and 682 nm), inverted microscopy, Trypan blue, LDH assay kit, ATP cell proliferation assay, FITC Annexin V/PI apoptosis kit.
4:Experimental Procedures and Operational Workflow:
Cells were treated with PSs and/or laser irradiation, followed by incubation. Changes were assessed using various assays.
5:Data Analysis Methods:
Statistical analysis was performed using Sigma Plot version 12.0.
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Hypericin
56690-1 MG
Sigma-Aldrich
Photosensitizer for photodynamic therapy
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Trypan blue
T8154
Sigma-Aldrich
Cell viability assay
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Aluminium(III) phthalocyanine chloride tetrasulphonate
Frontier Scientific
Photosensitizer for photodynamic therapy
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Diode laser
LREBT00 ROITHI
Oriel Corporation
Light source for activating photosensitizers
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Lactate Dehydrogenase Assay Kit
CytoTox96? assay, Promega G400
Promega
Measuring LDH released from cytosol into cell culture media
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CellTiter-Glo? Luminescent Assay
Promega, G7571
Promega
Evaluating metabolically active cells by measuring ATP signal
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FITC Annexin V Apoptosis Detection Kit
BD Pharmingen?, 556547
BD Pharmingen
Detecting early/late apoptotic or necrotic modes of cell death
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