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Apoptosis and necrosis induced with light and 5-aminolaevulinic acid-derived protoporphyrin IX
Open accessAbstract
The mode of cell death induced by photodynamic treatment (PDT) was studied in two cell lines cultured in monolayer, V79 Chinese hamster fibroblasts and WiDr human colon adenocarcinoma cells. The cells were incubated with 5-aminolaevulinic acid (5-ALA) as a precursor for the endogenously synthesised protoporphyrin IX, which was activated by light. Free DNA ends, owing to internucleosomal DNA cleavage in apoptotic cells, were stained specifically with a fluorescent dye in the terminal deoxynucleotidyl transferase (TdT) assay. The free DNA ends were measured by flow cytometry and the fractions of apoptotic cells determined. Total cell death was measured in a cell survival assay to determine the necrotic fraction after subtraction of the apoptotic fraction. V79 cells did undergo apoptosis while WiDr cells were killed only through necrosis. With time, the apoptotic fraction of V79 cells increased until a maximum was reached about 3-4 h after ALA-PDT treatment. For increasing ALA-PDT doses, a maximal apoptotic fraction 75-85% of the cells was measured at about 85% of total cell death. The flow cytometric assay of apoptosis was confirmed by the typical ladder of oligonucleosomal DNA fragments obtained from agarose gel electrophoresis, by fluorescence micrographs visualising the induced free DNA ends and by electron micrographs showing the typical morphology of apoptotic cells.
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“ALA-PDT has been shown to promote necrotic cell death as well, which is highly dependent on the cell line, PpIX localization, and PDT dose.”
From 5-Aminolevulinic Acid as a Theranostic Agent for Tumor Fluorescence Imaging and Photodynamic Therapy (2023) · cites this paper as [30]
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Metadata
- Authors
- Noodt, B., Berg, K., Stokke, T., Peng, Q., & Nesland, J.
- Affiliations
- Cancer Registry of Norway (NO)
- Venue
- British Journal of Cancer
- Published
- 1996-07
- Volume
- 74
- Issue
- 1
- Pages
- 22-29
- ISSN
- 0007-0920 (ISSN-L)
- 1532-1827
- PMC
- PMC2074599
- OpenAlex
- W1976916629
- Cited by
- 174
- References
- 38
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 3 · 2023: 5 · 2022: 5 · 2021: 2 · 2020: 3 · 2019: 2 · 2018: 4 · 2017: 7 · 2016: 3 · 2015: 4 · 2014: 5 · 2013: 5 · 2012: 2
- FWCI
- 6.46
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- British Journal of Cancer (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Photodynamic Therapy Research Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Porphyrin Metabolism and Disorders
- Keywords
- photodynamic therapy, 5-aminolevulinic acid, protoporphyrin IX, apoptosis, necrosis, cell death mode, V79 fibroblasts, DNA fragmentation, light activation
- MeSH terms
- Adenocarcinoma, Aminolevulinic Acid, Animals, Cells, Cultured, Colonic Neoplasms, Cricetulus, Fibroblasts, Cricetinae, Humans, Light, Necrosis, Photochemotherapy, Protoporphyrins, Tumor Cells, Cultured, Apoptosis, Photosensitizing Agents
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