Abstract
We have previously shown in LLC-PK1 cells, that apical membrane enzyme activity was inhibited by aminoglycoside antibiotics (Am. J. Physiol. 254, C251-C257, 1988). In the present study, the relationship between the lethal cytotoxic effect of aminoglycoside and its effect on apical membrane enzyme was examined by establishing aminoglycoside resistant cells. A clonal cell line, LLC-PK1/NRa3, was isolated from parent LLC-PK1 cells in the presence of neomycin. Neomycin inhibited colony formation and increased the number of floating dead cells in parent LLC-PK1 cultures. In contrast, these cytotoxic effects of neomycin were negligible or less pronounced in NRa3 cells, indicating that NRa3 cells were more resistant to neomycin compared with the parent cells. The inhibitory effect of neomycin on apical enzyme activity was significantly weaker in NRa3 cells than in the parent cells. These results suggest that a common mechanism is involved in the aminoglycoside-induced reductions in the apical enzyme activity and in cell viability of LLC-PK1 cells.
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REFERENCES
G. J. Kaloyanides and E. Pastoriza-Munoz. Aminoglycoside nephrotoxicity. Kidney Int. 18:571–582 (1980).
H. D. Humes and R. P. O'Connor. Aminoglycoside nephrotoxicity. In R. W. Schrier and C. W. Gottschalk (eds.), Diseases of the Kidney, 4th ed., Little, Brown, Boston, 1988, Vol. 2, pp. 1229–1273.
J. P. Morin, G. Viotte, A. Vandewalle, F. Van Hoof, P. Tulkens, and J. P. Fillastre. Gentamicin-induced nephrotoxicity: A cell biology approach. Kidney Int. 18:583–590 (1980).
H. Saito, K. Inui, and R. Hori. Mechanisms of gentamicin transport in kidney epithelial cell line (LLC-PK1). J. Pharmacol. Exp. Ther. 238:1071–1076 (1986).
C. A. Rabito, J. I. Kreisberg, and D. Wight. Alkaline phosphatase and γ-glutamyl transpeptidase as polarization markers during the organization of LLC-PK1 cells into an epithelial membrane. J. Biol. Chem. 259:574–582 (1984).
K. Inui, H. Saito, M. Takano, T. Okano, S. Kitazawa, and R. Hori. Enzyme activities and sodium-dependent active D-glucose transport in apical membrane vesicles isolated from kidney epithelial cell line (LLC-PK1). Biochim. Biophys. Acta 769:514–518 (1984).
D. S. Misfeldt and M. J. Sanders. Transepithelial transport in cell culture: Stoichiometry of Na/phlorizin binding and Na/D-glucose cotransport. A two-step, two-sodium model of binding and translocation. J. Membr. Biol. 70:191–198 (1982).
A. Moran, J. S. Handler, and R. J. Turner. Na+-dependent hexose transport in vesicles from cultured renal epithelial cell line. Am. J. Physiol. 243:C293–C298 (1982).
C. A. Rabito and M. V. Karish. Polarized amino acid transport by an epithelial cell line of renal origin (LLC-PK1): The apical systems. J. Biol. Chem. 258:2543–2547 (1983).
J. Caverzasio, C. D. A. Brown, J. Biber, J.-P. Bonjour, and H. Murer. Adaptation of phosphate transport in phosphate-deprived LLC-PK1 cells. Am. J. Physiol. 248:F122–F127 (1985).
A.-K. Fouda, C. Fauth, and F. Roch-Ramel. Transport of organic cations by kidney epithelial cell line LLC-PK1. J. Pharmacol. Exp. Ther. 252:286–292 (1990).
K. Inui, H. Saito, and R. Hori. H+-gradient-dependent active transport of tetraethylammonium cation in apical-membrane vesicles isolated from kidney epithelial cell line LLC-PK1. Biochem. J. 227:199–203 (1985).
H. Saito, M. Yamamoto, K. Inui, and R. Hori. Transcellular transport of organic cation across monolayers of kidney epithelial cell line LLC-PK1. Am. J. Physiol. 262:C59–C66 (1992).
K. Inui, H. Saito, T. Iwata, and R. Hori. Aminoglycoside-induced alterations in apical membranes of kidney epithelial cell line (LLC-PK1). Am. J. Physiol. 254:C251–C257 (1988).
R. Hori, K. Yamamoto, H. Saito, M. Kohno, and K. Inui. Effect of aminoglycoside antibiotics on cellular functions of kidney epithelial cell line (LLC-PK1): A model system for aminoglycoside nephrotoxicity. J. Pharmacol. Exp. Ther. 230:742–748 (1984).
R. Hori and K. Inui. Cellular basis of aminoglycoside nephrotoxicity. News Physiol. Sci. 4:181–184 (1989).
A. Wohlwend, J.-D. Vassalli, D. Belin, and L. Orci. LLC-PK1 cells: Cloning of phenotypically stable subpopulations. Am. J. Physiol. 250:C682–C687 (1986).
Y. Yoneyama and J. E. Lever. Increased trehalase expression after glucose limitation of LLC-PK1 clones. Am. J. Physiol. 255:C816–C821 (1988).
J. G. Haggerty, N. Agarwal, E. J. Cragoe, Jr., E. A. Adelberg, and C. W. Slayman. LLC-PK1 mutant with increased Na+ + H+ exchange and decreased sensitivity to amiloride. Am. J. Physiol. 255:C495–C501 (1988).
T. C. Knauss, J. M. Weinberg, and H. D. Humes. Alterations in renal cortical phospholipid content induced by gentamicin: Time course, specificity, and subcellular localization. Am. J. Physiol. 244:F535–F546 (1983).
M. Levi and R. E. Cronin. Early selective effects of gentamicin on renal brush-border membrane Na-Pi cotransport and Na-H exchange. Am. J. Physiol. 258:F1379–F1387 (1990).
L. S. Ramsammy, C. Josepovitz, and G. J. Kaloyanides. Gentamicin inhibits agonist stimulation of the phosphatidylinositol cascade in primary cultures of rabbit proximal tubular cells and in rat renal cortex. J. Pharmacol. Exp. Ther. 247:989–996 (1988).
L. S. Ramsammy, C. Josepovitz, B. Lane, and G. J. Kaloyanides. Effect of gentamicin on phospholipid metabolism in cultured rabbit proximal tubular cells. Am. J. Physiol. 256:C204–C213 (1989).
P. D. Walker and S. V. Shah. Evidence suggesting a role for hydroxyl radical in gentamicin-induced acute renal failure in rats. J. Clin. Invest. 81:334–341 (1988).
W. C. Elliott, D. C. Houghton, D. N. Gilbert, J. Baines-Hunter, and W. M. Bennett. Gentamicin nephrotoxicity. I. Degree and permanence of acquired insensitivity. J. Lab. Clin. Med. 100:501–512 (1982).
D. N. Gilbert, D. C. Houghton, W. M. Bennett, C. E. Plamp, K. Reger, and G. A. Porter. Reversibility of gentamicin nephrotoxicity in rats: Recovery during continuous drug administration. Proc. Soc. Exp. Biol. Med. 160:99–103 (1979).
M. Okuda, M. Takano, M. Yasuhara, and R. Hori. Inhibition of apical membrane enzyme activities and protein synthesis by gentamicin in a kidney epithelial cell line LLC-PK1. Chem. Pharm. Bull. 40:3307–3310 (1992).
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Hori, R., Okuda, M., Ohishi, Y. et al. Decreased Cellular Toxicity of Neomycin in a Clonal Cell Line Isolated from LLC-PK1 . Pharm Res 10, 573–576 (1993). https://doi.org/10.1023/A:1018954204094
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DOI: https://doi.org/10.1023/A:1018954204094