Metallothionein cell culture

Cherian, M.G. (1982). Studies on toxicity of metallothionein in rat kidney epithelial cell culture. Dev. Toxicol. Environ. Set 9 193-202. 

Liu J, Liu Y, Klaassen CD. Nephrotoxicity of CdCI2 and Cd-metallothionein in cultured rat kidney proximal tubules and LLC-PK1 cells. Toxicol AppI Pharmacol 1994 128 264-270. 

Despite the lack of a clear understanding of the mechanism by which metallothionein engenders drug resistance, the isotypes involved, and the subcellular location of metallothionein, several investigators have begun to probe whether the successful cell culture observations can be applied to preclinical tumor models or patients. 

Liu, J., Kershaw, W.C. and Klaasen, C.D. (1990). Rat primary hepatocyte cultures are a good model for examining metallothionein-induced tolerance to cadmium toxicity. In Vitro Cell. Dev. Biol. 26 75-79. 

Wolff NA, Abouhamed M, Verroust PJ, and Thevenod F. Megalln-dependent Internalization of cadmium-metallothionein and cytotoxicity In cultured renal proximal tubule cells. J Pharmacol ExpTher 318 782-791,2006. 

Lazo, J., Kondo, Y, DeUapiazza, D., Michalska, A., Choo, K., and Pitt, B. R. (1995). Enhcmced sensitivity to oxidative stress in cultured embryonic cells from transgenic mice different in metallothionein I and II genes. /. Biol. Chem. 27,5506-5510. 

Olsson, P.E., S.J. Hyllner, M. Zafarullah, T. Andersson and L. Gedamu. Differences in metallothionein expression in primary cultures of rainbow trout hepatocytes and the RTH-149 cell line. Biochim. Biophys. Acta 1049 78-82, 1990. 

Hyllner, S.J., T. Andersson, C. Haux and P.-E. Olsson. Cortisol induction of metallothionein in primary cultures of rainbow trout hepatocytes. J. Cell. Physiol. 239 24-28, 1989. 

Ishido M, Tohyama C, Suzuki T. Cadmium-bound metallothionein induces apoptosis in rat kidneys, but not in cultured kidney LLC-PK1 cells. Life Sci 1999 64 797-804. 

Dalton TP, Paria BC, Fernando LP, Huet-Hudson YM, Dey SK and Andrews GK (1997) Activation of the chicken metallothionein promoter by metals and oxidative stress in cultured cells and transgenic mice. Comp Biochem Physiol B Biochem Mol Biol 116 75 -86. 

Yao, C. P. Allen, J. W. Aschner, M. Metallothioneins Attenuate Methylmercury-induced Neurotoxicity in Cultured Astrocytes and Astrocytoma Cells. In Neuroprotective Agents Fourth International Conference-, Trembly, B., Slikker, W., Jr., Eds. New York, 1999 Vol. 890,... 

Dumam, D.M., Pahniter, R.D. 1984. Induction of metallothionein-I mRNA in cultured cells by heavy metals and iodoacetate evidence for gratuitous inducers. Mol. Cell. Biol. 4(3) 484—491. 

Various studies, both in cultured cells and in tissues from Menkes s disease patients and mottled mice mutants, have shown excess copper being accumulated in a low molecular weight cytoplasmic protein. All available evidence suggests that it is metallothionein. However, it is unclear as to whether its high concentration is a primary defect or whether it is induced by copper accumulation. It is a paradox that excess copper is being stored but enzymes requiring copper are unable to utilize the metal. 

NO mediates zinc release from the zinc-storage protein metallothionein via S-nitrosation and subsequent formation of disulphides (Kroncke et al. 1994). Culture of rat islet cells for 24 h in the presence of nontoxic concentrations (0.5 mM) of the slow-releasing NO donor diethylenetriamine/NO resulted in a significantly reduced Zn -dependent Zinquin (25 [iM) fluorescence (Tartler et al. 2000). 

Metallothionein expression in certain tumour cells has been associated with resistance to anticancer drugs. Increased resistance to chlorambucil occurred in cultured cells with a high concentration of cytoplasmic metallothionein (Endresen et al. 1983). Metallothionein inhibited hydroxyl radicalgenerated DNA degradation (Abel and Ruiter 1989). Satoh et al. (1994) showed that pretreatment of tumour-bearing ICR nude mice with zinc salts increased metallothionein content, both in normal and tumour tissues, with a marked reduction in the antitumour activity of cisplatin, Ad-riamycin , and melphalan. Metallothionein null cells have increased sensitivity to anticancer drugs (Kondo et al. 1995). 

Wan M, Hunziker PE, Kagi JH (1993) Induction of metallothionein synthesis by cadmium and zinc in cultured rabbit kidney cells (RK-13). Biochem J 292 609-615... 

Fernando LP, Andrews GK (1989) Cloning and expression of an avian metal-lothionein-encoding gene. Gene 81 177-183 Freedman JH, Peisach J (1989) Resistance of cultured hepatoma cells to copper toxicity. Purification and characterization of the hepatoma metallothionein. Biochim Biophys Acta 992 145-154... 

Ochi T (1988) Effects of glutathione depletion and induction of metallothioneins on the cytotoxicity of an orgainc hydroperoxide in cultured mammalian cells. Toxicology 50 257-268... 

Riordan JR, Richards V (1980) Human fetal liver contains both copper-rich forms of metallothionein. J Biol Chem 255 5380-5383 Robbins AH, McRae DE, Williamson M, Collett SA, Xuong NH, Furey WF, Wang BC, Stout CD (1991) Refined crystal structure of Cd.Zn metallothionein at 2.0 A resolution. J Mol Biol 221 1269-1293 Rugstad HE, Norseth T (1975) Cadmium resistance and content of cadmium-binding protein in cultured human cells. Nature 257 136 Ryden L, Deutsch HF (1978) Preparation and properties of the major copperbinding component in human fetal liver. J Biol Chem 253 519-524 Sato M, Bremner I (1993) Oxygen free radicals and metallothionein. Free Radic Biol Med 14 325-337... 

Coogan TP, Bare RM, Waalkes MP (1992) Cadmium-induced DNA damage effects of zinc pretreatment. Toxicol Appl Pharmacol 113 227-233 Coogan TP, Bare RM, Bjornson EJ, Waalkes MP (1994) Enhanced metallothionein gene expression protects against cadmium genotoxicity in cultured rat liver cells. J Toxicol Environ Health (in press)... 

Scand J Work Environ Health 12 523-537 Oberddrster G, Cox C (1990) Carcinogenicity of cadmium in animals what is the significance for man Chem Environ Toxicol 27 181-195 Ochi T, Ishiguro T, Ohsawa M (1983) Participation of active oxygen species in the induction of DNA single-strand scissions by cadmium chloride in cultured Chinese hamster cells. Mutat Res 122 169-175 Ochi T, Otsuka F, Takahashi K, Ohsawa M (1988) Glutathione and metallothioneins as cellular defense against cadmium toxicity in cultured Chinese hamster cells. Chem Biol Interact 65 1-14... 

Bunch TA, Grinblat Y, Goldstein LS (1988) Characterization and use of the Drosophila metallothionein promoter in cultured Drosophila melanogaster cells. Nucleic Acids Res 16 1043-1061... 

Bahnson RR, Banner BF, Ernstoff MS, Lazo JS, Cherian MG, Banerjee D, Chin JL (1991) Immunohistochemical localization of metallothionein in transitional cell carcinoma of the bladder. J Urol 146 1518-1520 Bakka A, Endresen L, Johnsen AB, Edminson PD, Rugstad HE (1981) Resistance against cis-dichlorodiammineplatinum in cultured cells with a high content of metallothionein. Toxicol Appl Pharmacol 61 215-226 Banerjee D, Onosaka S, Cherian MG (1982) Immunohistochemical localization of metallothionein in cell nucleus and cytoplasm of rat liver and kidney. Toxicology 24 95-105... 

Endresen L, Rugstad HE (1987) Protective function of metallothionein against certain anticancer agents. Experientia 52 595-602 Endresen L, Bakka A, Rugstad HE (1983) Increased resistance to chlorambucil in cultured cells with a high concentration of cytoplasmic metallothionein. Cancer Res 43 2918-2926... 

Studies on mutagenesis by metal compounds may be complicated by inducible tolerance mechanisms in some cells. Metallothioneins (MT) are small cysteine-rich proteins which bind a number of metals with high affinity. The reader is referred to Chap. 5 in this volume for more detail. Besides cadmium, zinc, and copper salts, a number of other metal salts have also been shown to induce MT synthesis and/or to bind to the MT protein. Hg(II), Co(II), and Ni(II), but not Pb(II), induce MT synthesis in primary cultures of rat hepatocytes (Bracken and Klaassen 1987), and may do so in the cultured cells used for mammalian mutagenesis experiments. Thus, under some experimental protocols, the metal salt being assayed may itself induce metallothionein, or the results of mutagenicity assays may be confounded by components of the medium or serum which affect the levels of metallothionein in the cell (Rossman and Goncharova, unpublished). 

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