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Molecules, Vol. 23, Pages 1575: Green Fluorescent Protein-Based Viability Assay in a Multiparametric Configuration

Molecules, Vol. 23, Pages 1575: Green Fluorescent Protein-Based Viability Assay in a Multiparametric Configuration

Molecules doi: 10.3390/molecules23071575

Authors: Rita Csepregi Viktória Temesfői Miklós Poór Zsuzsanna Faust Tamás Kőszegi

Green fluorescent protein (GFP) is considered to be suitable for cell viability testing. In our study, GFP transfected A549 lung carcinoma cell line was treated with sodium fluoride (NaF), cycloheximide (CHX) and ochratoxin A (OTA). GFP fluorescence, intracellular ATP, nucleic acid and protein contents were quantified by a luminescence microplate assay developed in our laboratory. Flow cytometry was used to confirm the findings and to assess the intensity of GFP during different types of cell death. A 24 h NaF and CHX exposure caused a dramatic decrease in ATP contents (p < 0.05) compared with those of the controls. GFP fluorescence of the cells was in close correlation with total protein; however, GFP/ATP increased at NaF and decreased at CHX treatments (p < 0.05). ATP/protein and ATP/propidium iodide (PI) were largely decreased at NaF exposure in a dose-dependent manner (p < 0.05), while CHX and OTA showed markedly fewer effects. Both treatments caused apoptosis/necrosis at different rates. NaF induced mainly late apoptosis while OTA, mainly apoptosis. CHX effects varied by the incubation time with 100-fold elevation in late apoptotic cells at 24 h treatment. GFP intensity did not show a significant difference between live and apoptotic populations. Our results suggest when using GFP, a multiparametric assay is necessary for more precise interpretation of cell viability.

Authors:   Csepregi, Rita ; Temesfői, Viktória ; Poór, Miklós ; Faust, Zsuzsanna ; Kőszegi, Tamás
Journal:   Molecules
Volume:   23
edition:   7
Year:   2018
Pages:   1575
DOI:   10.3390/molecules23071575
Publication date:   28-Jun-2018
Facts, background information, dossiers
  • NAF
  • molecules
  • green fluorescent protein
  • cells
  • cell viability
  • apoptosis
  • Cycloheximide
  • cell death
  • carcinoma
More about Molecular Diversity Preservation International
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