A transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress

Molina, A.; Carpeaux, Rudy; Martial, Joseph; Muller, Marc

doi:10.1016/S0887-2333(01)00106-0

Article (Scientific journals)

A transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress

Molina, A.; Carpeaux, Rudy; Martial, Joseph et al.

2002 • In Toxicology in Vitro, 16 (2), p. 201-7

Peer Reviewed verified by ORBi

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https://hdl.handle.net/2268/13023

DOI
10.1016/S0887-2333(01)00106-0

PubMed
11869883

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Keywords :

Animals; Cell Line, Transformed/drug effects/*metabolism; Dose-Response Relationship, Drug; Flow Cytometry; Genes, Reporter; Green Fluorescent Proteins; HSP70 Heat-Shock Proteins/genetics/metabolism; Indicators and Reagents/*metabolism; Luciferases/genetics/*metabolism; Luminescent Proteins/genetics/*metabolism; Recombinant Fusion Proteins/genetics/*metabolism; Tilapia; Xenobiotics/toxicity

Abstract :

[en] 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.

Research Center/Unit :

AFFISH-RC - Applied and Fundamental FISH Research Center - ULiège
Giga-Development and Stem Cells - ULiège

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Molina, A.

Carpeaux, Rudy ; Université de Liège - ULiège > Département des sciences cliniques > GIGA-R : Hématologie

Martial, Joseph ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire

Muller, Marc ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire

Language :

English

Title :

A transformed fish cell line expressing a green fluorescent protein-luciferase fusion gene responding to cellular stress

Publication date :

2002

Journal title :

Toxicology in Vitro

ISSN :

0887-2333

eISSN :

1879-3177

Publisher :

Pergamon-Elsevier Science Ltd, Oxford, United Kingdom

Volume :

Issue :

Pages :

201-7

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 22 May 2009

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Bibliography

Aït-Aïssa S., Porcher J., Arrigo A., Lambre C. (2000) Activation of the hsp70 promoter by environmental inorganic and organic chemicals: Relationships with cytotoxicity and lipophilicity. Toxicology 145:147-157.
Arai A., Mitani H., Naruse K., Shima A. (1994) Relationship between the induction of proteins in the HSP70 family and thermosensitivity in two species of Oryzias (Pisces). Comparative Biochemistry and Physiology-B. Biochemistry and Molecular Biology 109:647-654.
Brasier A.R., Tate J.E., Habener J. (1989) F Optimized use of the firefly luciferase assay as a reporter gene in mammalian cell lines. Biotechniques 7:1116-1122.
Day R.N., Kawecki M., Berry D. (1998) Dual-function reporter protein for analysis of gene expression in living cells. Biotechniques 25:848-856.
De Wachter B., Scholliers A., Blust R. (1998) Semiquantitative immunoblot detection of 70 kDa stress proteins in the carp Cyprinus carpio. Bulletin of Environmental Contamination and Toxicology 60:37-44.
Dunlap D.Y., Matsumura F. (1997) Development of broad-spectrum antibodies to heat shock protein 70s as biomarkers for detection of multiple stress by pollutants and environmental factors. Ecotoxicology and Environmental Safety 37:238-244.
Fijan N., Sulimanavoic D., Bearzotti M., Muzinic M.D., Zwilenberg L.O., Chilmonzyk S., Vautherot J.F., Kinkelin P. (1983) Some properties of the Epithelioma papulosum cyprini (EPC) cell line from carp Cyprinus carpio. Annales de l'Institut Pasteur/Virology 134:207-220.
Fischbach M., Sabbioni E., Bromley P. (1993) Induction of the human growth hormone gene placed under human hsp70 promoter control in mouse cells: A quantitative indicator of metal toxicity. Cell Biology and Toxicology 9:177-188.
Freeman M.L., Borelli M.J., Meredith M.J., Lepock J.R. (1999) On the path to the heat shock response: Destabilization and formation of partially folded protein intermediates, a consequence of protein thiol modification. Free Radical Biology and Medicine 145:147-157.
Han S.I., Oh S.Y., Woo S.H., Kim K.H., Kim J.-H., Kim H.D., Kang H.S. (2001) Implication of a small GTPase Rac1 in the activation of c-Jun terminal kinase and heat shock factor in response to heat shock. Journal of Biological Chemistry 276:1889-1895.
Hartl U.F. (1996) Molecular chaperones in cellular protein folding. Nature 381:571-580.
Jaattela M. (1999) Escaping cell death: Survival proteins in cancer. Experimental Cell Research 248:30-43.
Krone P.H., Heikkila J.J. (1988) Analysis of hsp 30, hsp 70 and ubiquitin gene expression in Xenopus laevis tadpoles. Development 103:59-67.
Misra S., Zafarullah M., Price-Haughey J., Gedamu L. (1989) Analysis of stress-induced gene expression in fish cell lines exposed to heavy meals and heat shock. Biochimica et Biophysica Acta 1007:325-333.
Molina A., Biemar F., Muller F., Iyengar A., Prunet P., Maclean N., Martial J.A., Muller M. (2000) Cloning and expression analysis of an inducible HSP70 gene from tilapia fish. FEBS Letters 474:5-10.
Molina A., Di Martino E., Martial J.A., Muller M. (2001) Heat shock stimulation of a tilapia fish HSP70 promoter is mediated by a distal element. Biochemical Journal 356:353-359.
Morimoto R.I. (1993) Cells in stress: Transcriptional activation of heat shock genes. Science 259:1409-1410.
Morimoto R.I. (1998) Regulation of the heat shock transcriptional response: Cross talk between a family of heat shock factors, molecular chaperones, and negative regulators. Genes and Development 2:3788-3796.
Neuhaus-Steinmetz U., Rensing L. (1997) Heat shock protein induction by certain chemical stressors is correlated with their cytotoxicity, lipophilicity and protein-denaturing capacity. Toxicology 123:185-195.
Rabindran S.K., Haroun R.I., Clos J., Wisniewski J., Wu C. (1993) Regulation of heat shock factor trimer formation: Role of a conserved leucine zipper. Science 259:230-234.
Ritossa F. (1962) A new puffing pattern induced by heat shock and DNP in Drosophila. Experientia 18:571-573.
Samali A., Holmberg C.I., Sistonen L., Orrenius S. (1999) Thermotolerance and cell death are distinct cellular responses to stress: Dependence on heat shock proteins. FEBS Letters 461:306-310.
Sistonen L., Sarge K.D., Morimoto R.I. (1994) Human heat shock factors 1 and 2 are differentially activated and can synergistically induce hsp70 gene transcription. Molecular and Cellular Biology 14:2087-2099.
Steiner E., Kleinhappl B., Gutschi A., Marth E. (1998) Analysis of hsp70 mRNA levels in HepG2 cells exposed to various metals differing in toxicity. Toxicology Letters 96-97:169-176.
Todd M.D., Lee M.J., Williams J.L., Nalezny J.M., Gee P., Benjamin M.B., Farr S.B. (1995) The CAT-Tox (L) assay: A sensitive and specific measure of stress-induced transcription in transformed human liver cells. Fundamental and Applied Toxicology 28:118-128.
Vincent R., Goegan P., Johnson G., Brook J.R., Kumarathasan P., Bouthillier L., Burnett R.T. (1997) Regulation of promoter-CAT stress genes in HepG2 cells by suspensions of particles from ambient air. Fundamental and Applied Toxicology 39:18-32.
Vogel P., Dux E., Wiessner C. (1997) Effect of heat shock on neuronal cultures: Importance of protein synthesis and HSP72 induction for induced tolerance and survival. Metabolic Brain Disease 12:203-217.