Kidney slices represent an in vitro model that has the cellular complexity of in vivo tissue to provide insights into mechanisms of organ injury, as shown in this study with the model nephrotoxicant cisplatin. Cell pathways altered by cisplatin exposure are assessed by gene expression analysis, cell function, and morphology in human and rat kidney slices in comparison to rat kidney from an in vivo study. The acute nephrosis of the tubular epithelium induced by cisplatin in vivo was reproduced in both human and rat kidney slices, while the glomerulus appeared resistant even at high concentrations. Kidney gene expression changes of in vivo and in vitro samples were indicative of transcription, DNA damage, cell cycle, proliferation, and apoptosis that are in agreement with the mechanism of cisplatin causing DNA damage, growth arrest, and apoptosis; while genes indicative of protein damage, the disruption of transport and calcium homeostasis, cellular metabolism, and oxidative stress are pathways linked with cisplatin binding to various cellular proteins and macromolecules. Both concentration and time-dependent gene expression changes evident in the in vitro model preceded a change in tissue morphology. Functional assays confirming cell dysfunction and increased apoptosis revealed the rat kidney to be more sensitive to the effects of cisplatin than human kidney as demonstrated by significant decreases in slice ATP and GSH levels, significant increases in caspase 9 and 3 activity, p53 protein levels, and increased DNA laddering. The regional markers of proximal and distal tubular injury, alpha- and pi-glutathione S-transferases, were shown for the human kidney slices to be significantly increased by cisplatin. In this study, cisplatin-induced nephrotoxicity was demonstrated morphologically in rat and human kidney slices, and the associated gene expression and functional changes characterized the cellular pathways involved.
Agarwal, A., Balla, J., Alam, J., Croatt, A.J., and Nath, K.A. (1995). Induction of heme oxygenase in toxic renal injury: A protective role in cisplatin nephrotoxicity in the rat. Kidney Int48, 1298—307.
2.
Campbell, J.A.H., Corrigal, A.V., Guy, A., and Kirsch, R.E. (1991). Immunohistologic localization of alpha, mu, and pi class glutathione S-transferases in human tissues. Cancer6, 1608—13.
3.
Chen, G., and Zeller, W.J. (1991). Augmentation of cisplatin (DDP) cytotoxicity in vivo by DL-buthionine sulfoximine (BSO) in DDP-sensitive and-resistant rat ovarian tumors and its relation to DNA interstrand cross links. Anticancer Res11, 2231—8.
4.
Choie, D.D., Longnecker, D.S., and Del Campo, A.A. (1981). Acute and chronic cisplatin nephropathy in rats . Lab Invest44, 397—402.
5.
Chopra, S., Kaufman, J.S., Jones, T.W., Hong, W.K., Gehr, M.K., Hamburger, R.J., Flamenbaum, W., and Trump, B.F. (1982). Cis-diamminedichloroplatinum-induced acute renal failure in the rat. Kidney Int21, 54—64.
6.
Dehne, N., Lautermann, J., Petrat, F., Rauen, U., and de Groot, H. (2001). Cisplatin ototoxicity: involvement of iron and enhanced formation of super-oxide anion radicals. Toxicol Appl Pharmacol174, 27—34.
7.
Delmastro, D.A., Li, J., Vaisman, A., Solle, M., and Chaney, S.G. (1997). DNA damage inducible-gene expression following platinum treatment in human ovarian carcinoma cell lines. Cancer Chemother Pharmacol39, 245— 53.
8.
Dentino, M., Luft, F.C., Yum, M.N., Williams, S.D., and Einhorn, L.H. (1978). Long term effect of cis-diamminedichloride platinum (CDDP) on renal function and structure in man. Cancer41, 1274—81.
9.
Eder, P. (1997). Neoplasms. In Integrated Pharmacology (C. P. Page, M. J. Curtis, M. C. Sutter, M. J. A. Walker, and B. B. Hoffman, eds.), pp. 501— 22. Mosby International, St. Louis, MO.
10.
Endo, T., Kimura, O., and Sakata, M. (2002). Carrier-mediated uptake of cisplatin by the OK renal epithelial cell line. Toxicology146, 187—95.
11.
Erdinc, M., Erdinc, L., Nergiz, Y., and Isik, B. (2000). Potentiation of cisplatin-induced nephrotoxicity in rats with allopurinol. Exp Toxicol Pathol52, 329—34.
12.
Fan, J., and Bertino, J.R. (1999). Modulation of cisplatinum cytotoxicity by p53: effect of p53-mediated apoptosis and DNA repair. Mol Pharmacol56, 966—72.
13.
Fisher, R.L., Sanuik, J.T., Gandolfi, A.J., and Brendel, K. (1994). Toxicity of cisplatin and mercuric chloride in human kidney cortical slices. Human Exper Toxicol13, 517—523.
14.
Gonzalez, V.M., Fuertes, M.A., Alonso, C., and Perez, J.M. (2001). Is cisplatin-induced cell death always produced by apoptosis? Mol Pharmacol59, 657—63.
15.
Gonzalez-Vitale, J.C., Hayes, D.M., Cvitkovic, E., and Sternberg, S.S. (1978). The renal pathology in clinical trials of cis-platinum (II) and gentamicincephalothin therapies. Cancer Treat Rep62, 693—8.
16.
Hagrman, D., Goodisman, J., Dabrowiak, J.C., and Souid, A.-K. (2003). Kinetic study on the reaction of cisplatin with metallothionein. Drug Metabol Dispos31, 916—23.
17.
Halabe, A., Wong, N.L.M., and Sutton, R.A.L. (1991). Effect of chronic cisplatin administration on phosphate and glucose transport by the renal brush border membrane. Nephron57, 197—200.
18.
Hardaker, W.T. Jr, Stone, R.A., and McCoy, R. (1974). Platinum nephrotoxicity. Cancer34, 1030—2.
19.
Hosokawa, T., Okabe, M., Saito, S., Saito, T., and Kurasaki, M. (2000). Protective role of metallothionein on DNA damage in rat kidney caused by cis-diamminedichloroplatinum. Pharmacol Toxicol86, 276— 82.
20.
Huang, Q., Dunn, R.T., Jayadev, S., DiSorbo, O., Pack, F.D., Farr, S.B., Stoll, R.E., and Blanchard, K.T. (2001). Assessment of cisplatin-induced nephrotoxicity by microarray technology. Toxicol Sci63, 196— 207.
21.
Ihnat, M.A., Lariviere, J.P., Warren, A.J., La Ronde, N., Blaxall, J.R.N., Pierre, K.M., Turpie, B.W., and Hamilton, J.W. (1997). Suppression of P-glycoprotein expression and multidrug resistance by DNA cross-linking agents. Clin Cancer Res3, 1339—46.
22.
Ishikawa, T., Wright, C.D., and Ishizuka, H. (1994). GS-X pump is functionally overexpressed in cis-diamminedichloroplatinum (II)-resistant human leukemia HL-60 cells and down-regulated by cell differentiation . J Biol Chem269, 29085—93.
23.
Johnstone, R.W., Ruefli, A.A., and Smyth, M.J. (2000). Multiple physiological functions for multidrug transporter P-glycoprotein? TIBS25, 1—6.
24.
Jones, N.A., Turner, J., McIlwrath, A.J., Brown, R., and Dive, C. (1998). Cisplatin- and paclitaxel-induced apoptosis of ovarian carcinoma cells and the relationship between bax and bak up-regulation and the functional status of p53. Mol Pharmacol53, 819—26.
25.
Leibbrandt, M.E., and Wolfgang, G.H. (1995). Differential toxicity of cisplatin, carboplatin, and CI-973 correlates with cellular platinum levels in rat cortical slices . Toxicol Appl Pharmacol132, 245—52.
26.
Leibbrandt, M.E., Wolfgang, G.H., Metz, A.L., Ozobia, A.A., and Haskins, J.R. (1995). Critical subcellular targets of cisplatin and related platinum analogs in rat proximal tubule cells. Kidney Int48, 761—70.
27.
Lieberthal, W., Menza, S.A., and Levine, J.S. (1998). Graded ATP depletion can cause necrosis or apoptosis of cultured mouse proximal tubular cells. Am J Physiol274, F315—27.
28.
Lieberthal, W., Triarca, V., and Levine, J. (1996). Mechanisms of death induced by cisplatin in proximal tubular epithelial cells: apoptosis vs. necrosis. Am J Physiol270, F700—8.
29.
Liu, J., Liu, Y., Habeebu, S.S.M., and Klaassen, C.D. (1998). Metallothionein (MT)-null mice are sensitive to cisplatin-induced hepatotoxicity. Toxicol Appl Pharmacol149, 24—31.
30.
Mandic, A., Viktorsson, K., Molin, M., Akusjarvi, G., Eguchi, H., Hayashi, S.I., Toi, M., Hansson, J., Linder, S., and Shoshan, M.C. (2001). Cisplatin induces the proapoptotic conformation of Bak in a AMEKK1-dependent manner. Mol Cell Biol21, 3684—91.
31.
Nagai, N., Okuda, M., Kinoshita, M., and Ogata, H. (1996). Decomposition kinetics of cisplatin in human biological fluids. J Pharm Pharmacol48, 918—24.
32.
Niedner, H., Christen, R., Lin, X., Kondo, L.A., and Howell, S.B. (2001). Identification of genes that mediate sensitivity to cisplatin. Mol Pharmacol60, 1153—60.
33.
O'Brien, T., Babcock, G., Cornelius, J., Dingeldein, M., Talaska, G., Warshawsky, D., and Mitchell, K. (2000). A comparison of apoptosis and necrosis induced by hepatotoxins in HepG2 cells. Toxicol Appl Pharmacol164, 280—90.
34.
Perez, R.P. (1998). Cellular and molecular determinants of cisplatin resistance. Eur J Cancer34, 1535—42.
35.
Piel, I.J., and Perlia, C.P. (1975). Phase II study of cisdichlorodiammineplatinum (II) (NS 119875) in combination with cyclophosphamide (NSC-26271) in the treatment of human malignancy. Cancer Chemother Rep59, 995—9.
36.
Plas, D.R., and Thompson, C.B. (2002). Cell metabolism in the regulation of programmed cell death. Trends Endocrinol Metabol13, 74—8.
37.
Rixe, O. (2000). Platinum salts: cytotoxic mechanisms of action, mechanisms of resistance of cancer cells, interactions with ionizing radiation, specificity of carboplatin. Bull Cancer87, 7—15.
38.
Sadowitz, P.D., Hubbard, B.A., Dabrowiak, J.C., Goodisman, J., Tacka, K.A., Aktas, K.M., Cunningham, M.J., Dubowy, R.L., and Souid, A.-K. (2002). Kinetics of cisplatin binding to cellular DNA and modulations by thiol-blocking agents and thiol drugs. Drug Metabol Disposit30, 183—90.
39.
Satoh, M., Shimada, A., Zhang, B., and Chiharu, T. (2000). Renal toxicity caused by cisplatinum in glutathione-depleted metallothionein-null mice. Biochem Pharmacol60, 1729—34.
40.
Sheikh, M.S., Hollander, M.C., and Fornace, A.J., Jr. (2000). Role of Gadd45 in apoptosis. Biochem Pharmacol59, 43—5.
41.
Shimoda, R., Achanzar, W.E., Qu, W., Nagamine, T., Takagi H., Mori, M., and Waalkes, M.P. (2003). Metallothionein is a potential negative regulator of apoptosis. Toxicol Sci73, 294—300.
42.
Shiraishi, F., Curtis, L.M., Truong, L., Poss, K., Visner, G.A., Madsen, K., Nick, H.S., and Agarwal, A. (2000). Heme oxygenase-1 gene ablation or expression modulates cisplatin-induced renal tubular apoptosis. Am J Physiol Renal Physiol278, F726—36.
43.
Silkensen, J.R., Agarwal, A., Nath, K.A., Manivel, J.C., and Rosenberg, M.E. (1997). Temporal induction of clusterin in cisplatin nephrotoxicity . J Am Soc Nephrol8, 302—5.
44.
Smith, M.L., Ford, J.M., Hollander, M.C., Bortnick, R.A., Amundson, S.A., Seo, Y.R., Deng, C.-X., Hanawalt, P.C., and Fornace, Jr., A.J. (2000). p53-Mediated DNA repair responses to UV radiation: studies of mouse cells lacking p53, p21, and/or gadd45 genes. Mol Cell Biol20, 3705— 14.
45.
Sundberg, A.G.M., Nilsson, R., Appelkvist, E.-L., and Dallner, G. (1993). Immunohistochemical localization of α and π class glutathione transferases in normal human tissues. Pharmacol Toxicol72, 321—31.
46.
Sundberg, A.G.M., Appelkvist, E.-L., Backham, L., and Dallner, G. (1994). Urinary π-class glutathione transferase as an indicator of tubular damage in the human kidney. Nephron67, 308—16.
47.
Tanaka, H., Ishikawa, E., Teshima, S., and Shimizu, E. (1986). Histopathological study of human cisplatin nephropathy . Toxicol Pathol14, 247—57.
48.
Thompson, K.L., Afshari, C.A., Amin, R.P., Bertram, T.A., Car, B., Cunningham, M., Kind, C., Kramer, J.A., Lawton, M., Mirsky, M., Naciff, J.M., Oreffo, V., Pine, P.S., and Sistare, F.D. (2004). Identification of platform-independent gene expression markers of cisplatin nephrotoxicity. Environ Hlth Perspec112, 488—94.
49.
Vasey, P.A., Jones, N.A., and Jenkins, S. (1996). Cisplatin, camptothecin, and taxol sensitivities of cells with p53-associated multidrug resistance. Mol Pharmacol50, 1536—40.
50.
Wang, X.W., Xhan, O., Coursen, J.D., Khan, M.A., Kontny, H.U., Yu, L., Hollander, M.C., O'Connor, P.M., Fornace, Jr., A.J., and Harris, C.C. (1999). GADD45 induction of a G2/M cell cycle checkpoint . Proc Natl Acad Sci USA96, 3706—11.
51.
Zhang, K., Chew, M., Yang, E.B., Wong, K.P., and Mack, P. (2001). Modulations of cisplatin cytotoxicity and cisplatin-induced DNA cross-links in HepG2 cells by regulation of glutathione-related mechanisms . Mol Pharmacol59, 837—43.
