A transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress.
We obtained a stable transformed fish (EPC) cell line containing a reporter gene under the control of the tilapia HSP70 promoter. Expression of the reporter gene, coding for a green fluorescent protein (GFP)-luciferase fusion protein, was assessed by measuring the luciferase enzymatic activity by luminometry and the GFP expression by fluorescence microscopy and flow cytometry. The clone was characterized for its capacity to respond to heat shock treatment. The results show high induction after 1 h at 37 degrees C of treatment, up to 500-fold. In addition, its convenience to detect a large range of cellular stressors was evaluated. We observed high induction when Cd2+, Zn2+, Hg2+ or Cu2+ was added, but not Pb2+. In addition, activation of the reporter gene was observed in the presence of other compounds such as acetyl chloride, tetrachlorophenol, chloroacetamide and sodium arsenite. In conclusion, this cell line can be used as a rapid, cheap and easy biological test to determine cellular stress induced by environmental pollutants, alone or in conjunction with other, more specific assays.
Molina
A
A
Laboratoire de Biologie Moléculaire et Génie Génétique, Université de Liège, Institut de Chimie B6, B-4000 Sart-Tilman, Belgium.
Carpeaux
R
R
Martial
J A
JA
Muller
M
M
D000818:Animals; D002461:Cell Line, Transformed / Q000187:drug effects / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D007202:Indicators and Reagents / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D017210:Tilapia; D015262:Xenobiotics / Q000633:toxicity
D018840:HSP70 Heat-Shock Proteins; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D015262:Xenobiotics; D049452:Green Fluorescent Proteins; D008156:Luciferases