Abstract
Very little is known about the modulation of vanadium accumulation in cells, although this ultratrace element has long been seen as an essential nutrient in lower life forms, but not necessarily in humans where factors modulating cellular uptake of vanadium seem unclear. Using nuclear microscopy, which is capable of the direct evaluation of free and bound (total) elemental concentrations of single cells we show here that an NH4Cl acidification prepulse causes distinctive accumulation of vanadium (free and bound) in human Chang liver cells, concentrating particularly in the nucleus. Vanadium loaded with acidification but leaked away with realkalinization, suggests proton-dependent loading. Vanadyl(4), the oxidative state of intracellular vanadium ions, is known to be a potent source of hydroxyl free radicals (OH.). The high oxidative state of nuclei after induction of vanadyl(4) loading was shown by the redox indicator methylene blue, suggesting direct oxidative damage to nuclear DNA. Flow cytometric evaluation of cell cycle phase-specific DNA composition showed degradation of both 2N and 4N DNA phases in G1, S and G2/M cell cycle profiles to a solitary 1N DNA peak, in a dose-dependent manner, effective from micromolar vanadyl(4) levels. This trend was reproduced with microccocal nuclease digestion in a time response, supporting the notion of DNA fragmentation effects. Several other approaches confirmed fragmentation occurring in virtually all cells after 4 mM V(4) loading. Ultrastructural profiles showed various stages of autophagic autodigestion and well defined plasma membrane outlines, consistent with programmed cell death but not with necrotic cell death. Direct intranuclear oxidative damage seemed associated with the induction of mass suicide in these human Chang liver cells following vanadium loading and nuclear sequestration.
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References
Chasteen, N. D., Structure and Bonding53 (1983) 105.
Carpenter, G., Biochem. Biophys. Res. Commun.102 (1981) 1115.
Sit, K. H., Paramanantham, R., Bay, B. H., and Wong, K. P., Jap. J. Pharmacol.65 (1994) 83.
Ishikawa, T., Trends Biochem. Sci.17 (1992) 463.
Muller, M., Meijer, C., Zaman, G. J. R., Borst, P., Scheper, R. J., Mulder, N. H., de Vries, E. G. E., and Jansen, P. L. M., Proc. natl Acad. Sci. USA91 (1994) 13033.
Posner, B. I., Faure, R., Burgess, J. W., Bevan, A. P., Lachance, D., Zhang-Sun, G., Fantus, I. G., Ng, J. B., Hall, D. A., Lum, B. S., and Shaver, A., J. biol. Chem.269 (1994) 4596.
Thompson, H. J., Chasteen, D. N., and Meeker, L. D., Carcinogenesis5 (1984) 849.
Matsumoto, J., Medical Hypothesis43 (1994) 177.
French, R. J., and Jones, P. J., Life Sci.52 (1994) 339.
Harland, B. F., and Harden-Williams, B. A., J. Am. Diet. Assoc.94 (1994) 891.
Sit, K. H., Bay, B. H., Paramanantham, R., Thong, P., and Watt, F., J. Tiss. Cult. Meth.15 (1993) 199.
Landsberg, J. P., McDonald, B., and Watt, F., Nature (Lond.)360 (1992) 65.
Watt, F., Orlic, I., Loh, K. K., Sow, C. H., Thong, P., Liew, S. C., Osipowicz, T., Choo, R. F., and Tang, S. M., Nucl. Instr. Meths.B85 (1994) 708.
Cargnello, J. A., Powell, J. J., Thompson, R. P. H., Crocker, P. R., and Watt, F., Analyst120 (1995) 783.
Hamada, T., Experientia15 (1994) 49.
Monteiro, H. P., Winterbourn, C. C., and Stern, A., Free Radic. Res. Commun.12–13 (1991) 125.
Keller, R. J., Sharma, R. P., Grover, T. A., and Piette, L. H., Archs Biochem. Biophys.265 (1988) 524.
Liochev, S. I., and Fridovich, I., Archs Biochem. Biophys.29 (1991) 379.
Kane, D. J., Sarafian, T. A., Anton, R., Hahn, H., Gralla, E. B., Valentine, J. S., Ord, T., and Bredesen, D. E., Science262 (1993) 1274.
Hockenbery, D. M., Oltvai, Z. N., Yin, X.-M., Milliman, C. L., and Korsmeyer, S. J., Cell75 (1993) 241.
Thompson, C., Science267 (1995) 1456.
Korsmeyer, S. J., Trends Genetics11 (1995) 101.
Hansard, S. L., Ammerman, C. B., Henry, P. R., and Simpson, C. F., J. Anim. Sci.55 (1982) 344.
Wei, C., Al Bayati, M. A., Culbertson, M. R., Rosenblatt, L. S., and Hansen, L. D., J. Toxicol. Environ. Health10 (1982) 673.
Lee, K. P., and Gillies, P. J., Environ. Res.40 (1986) 115.
Sit, K. H., and Wong, K. P., Tissue Cell21 (1989) 321.
Sit, K. H., Paramanatham, R., Bay, B. H., and Wong, K. P., Anat. Rec.240 (1994) 456.
Baker, M. S., and Gebicki, J. M., Archs Biochem. Biophys.234 (1984) 258.
Telford, W. G., King, L. E., and Fraker, P. J., Cytometry13 (1992) 137.
Umansky, S. R., in: Apoptosis: The Molecular Basis of Cell Death (Current Communications in Cell and Molecular Biology, 3), p. 193. Eds D. D. Tomei and F. O. Cope. Cold Spring Harbor Laboratory Press, New York 1991.
Telford, W. G., King, L. E., and Fraker, P. J., J. Immun. Meth.172 (1994) 1.
Halliwell, B., and Gutteridge, J. M. C., Biochem. J.219 (1984) 1.
Dunn, W. A. Jr., J. Cell Biol.110 (1990) 1935.
Lockshin, R. A., and Zakeri, Z., in: Apoptosis: The Molecular Basis of Cell Death (Current Communications in Cell and Molecular Biology, 3), p. 47. Eds D. D. Tomei and F. O. Cope. Cold Spring Harbor Laboratory Press, New York 1991.
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Sit, K.H., Paramanantham, R., Bay, B.H. et al. Induction of vanadium accumulation and nuclear sequestration causing cell suicide in human Chang liver cells. Experientia 52, 778–785 (1996). https://doi.org/10.1007/BF01923989
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DOI: https://doi.org/10.1007/BF01923989