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. 1997 Nov;17(11):6755–6764. doi: 10.1128/mcb.17.11.6755

A role for c-myc in chemically induced renal-cell death.

Y Zhan 1, J L Cleveland 1, J L Stevens 1
PMCID: PMC232530  PMID: 9343440

Abstract

A variety of genes, including c-myc, are activated by chemical toxicants in vivo and in vitro. Although enforced c-myc expression induces apoptosis after withdrawing survival factors, it is not clear if activation of the endogenous c-myc gene is an apoptotic signal after toxicant exposure. The renal tubular epithelium is a target for many toxicants. c-myc expression is activated by tubular damage. In quiescent LLC-PK1 renal epithelial cells, c-myc but not max or mad mRNA is induced by the nephrotoxicant S-(1,2-dichlorovinyl)-L-cysteine (DCVC). The kinetics of DCVC-induced c-myc expression and apoptosis suggested an association between cell death and prolonged activation of c-myc expression after toxicant exposure. Accordingly, prolonged activation of an estrogen receptor-Myc fusion construct, but not a construct in which a c-Myc transactivation domain had been deleted, was sufficient to induce apoptosis in LLC-PK1 cells. Moreover, under conditions in which necrosis was the predominant cell death pathway caused by DCVC in parental cells, overexpressing c-myc biased the cell death pathway toward apoptosis. DCVC also induced ornithine decarboxylase (odc) mRNA and activated the odc promoter. Activation of the odc promoter by DCVC required consensus c-Myc-Max binding sites in odc intron 1. Inhibiting ODC activity with alpha-difluoromethylornithine delayed DCVC-induced cell death. Therefore, odc is a target gene in the DCVC apoptotic pathway involving c-myc activation and contributes to apoptosis. Finally, a structurally related cytotoxic but nongenotoxic analog of DCVC did not induce c-myc and did not activate the odc promoter or induce apoptosis. The data support the hypothesis that activation of apoptotic cell death in quiescent renal epithelial cells involves induction of c-myc. This is the first study to demonstrate that c-myc induction by a specific nephrotoxicant leads to gene activation and cell death.

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Selected References

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  1. Askew D. S., Ashmun R. A., Simmons B. C., Cleveland J. L. Constitutive c-myc expression in an IL-3-dependent myeloid cell line suppresses cell cycle arrest and accelerates apoptosis. Oncogene. 1991 Oct;6(10):1915–1922. [PubMed] [Google Scholar]
  2. Ayer D. E., Kretzner L., Eisenman R. N. Mad: a heterodimeric partner for Max that antagonizes Myc transcriptional activity. Cell. 1993 Jan 29;72(2):211–222. doi: 10.1016/0092-8674(93)90661-9. [DOI] [PubMed] [Google Scholar]
  3. Ayer D. E., Lawrence Q. A., Eisenman R. N. Mad-Max transcriptional repression is mediated by ternary complex formation with mammalian homologs of yeast repressor Sin3. Cell. 1995 Mar 10;80(5):767–776. doi: 10.1016/0092-8674(95)90355-0. [DOI] [PubMed] [Google Scholar]
  4. Basu A., Cline J. S. Oncogenic transformation alters cisplatin-induced apoptosis in rat embryo fibroblasts. Int J Cancer. 1995 Nov 15;63(4):597–603. doi: 10.1002/ijc.2910630422. [DOI] [PubMed] [Google Scholar]
  5. Beijersbergen R. L., Hijmans E. M., Zhu L., Bernards R. Interaction of c-Myc with the pRb-related protein p107 results in inhibition of c-Myc-mediated transactivation. EMBO J. 1994 Sep 1;13(17):4080–4086. doi: 10.1002/j.1460-2075.1994.tb06725.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bello-Fernandez C., Packham G., Cleveland J. L. The ornithine decarboxylase gene is a transcriptional target of c-Myc. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7804–7808. doi: 10.1073/pnas.90.16.7804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Berberich S. J., Cole M. D. Casein kinase II inhibits the DNA-binding activity of Max homodimers but not Myc/Max heterodimers. Genes Dev. 1992 Feb;6(2):166–176. doi: 10.1101/gad.6.2.166. [DOI] [PubMed] [Google Scholar]
  8. Bernards R. Transcriptional regulation. Flipping the Myc switch. Curr Biol. 1995 Aug 1;5(8):859–861. doi: 10.1016/s0960-9822(95)00173-4. [DOI] [PubMed] [Google Scholar]
  9. Bissonnette R. P., Echeverri F., Mahboubi A., Green D. R. Apoptotic cell death induced by c-myc is inhibited by bcl-2. Nature. 1992 Oct 8;359(6395):552–554. doi: 10.1038/359552a0. [DOI] [PubMed] [Google Scholar]
  10. Blackwood E. M., Eisenman R. N. Max: a helix-loop-helix zipper protein that forms a sequence-specific DNA-binding complex with Myc. Science. 1991 Mar 8;251(4998):1211–1217. doi: 10.1126/science.2006410. [DOI] [PubMed] [Google Scholar]
  11. Chen Q., Jones T. W., Brown P. C., Stevens J. L. The mechanism of cysteine conjugate cytotoxicity in renal epithelial cells. Covalent binding leads to thiol depletion and lipid peroxidation. J Biol Chem. 1990 Dec 15;265(35):21603–21611. [PubMed] [Google Scholar]
  12. Cherney B. W., Bhatia K., Tosato G. A role for deregulated c-Myc expression in apoptosis of Epstein-Barr virus-immortalized B cells. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12967–12971. doi: 10.1073/pnas.91.26.12967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cowley B. D., Jr, Chadwick L. J., Grantham J. J., Calvet J. P. Sequential protooncogene expression in regenerating kidney following acute renal injury. J Biol Chem. 1989 May 15;264(14):8389–8393. [PubMed] [Google Scholar]
  14. Davidoff A. N., Mendelow B. V. Puromycin-elicited c-myc mRNA superinduction precedes apoptosis in HL-60 leukaemic cells. Anticancer Res. 1993 Nov-Dec;13(6A):2257–2260. [PubMed] [Google Scholar]
  15. Davis R. H., Morris D. R., Coffino P. Sequestered end products and enzyme regulation: the case of ornithine decarboxylase. Microbiol Rev. 1992 Jun;56(2):280–290. doi: 10.1128/mr.56.2.280-290.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dekant W., Vamvakas S., Koob M., Köchling A., Kanhai W., Müller D., Henschler D. A mechanism of haloalkene-induced renal carcinogenesis. Environ Health Perspect. 1990 Aug;88:107–110. doi: 10.1289/ehp.9088107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Evan G. I., Littlewood T. D. The role of c-myc in cell growth. Curr Opin Genet Dev. 1993 Feb;3(1):44–49. doi: 10.1016/s0959-437x(05)80339-9. [DOI] [PubMed] [Google Scholar]
  18. Fanidi A., Harrington E. A., Evan G. I. Cooperative interaction between c-myc and bcl-2 proto-oncogenes. Nature. 1992 Oct 8;359(6395):554–556. doi: 10.1038/359554a0. [DOI] [PubMed] [Google Scholar]
  19. Galaktionov K., Chen X., Beach D. Cdc25 cell-cycle phosphatase as a target of c-myc. Nature. 1996 Aug 8;382(6591):511–517. doi: 10.1038/382511a0. [DOI] [PubMed] [Google Scholar]
  20. Green T., Odum J. Structure/activity studies of the nephrotoxic and mutagenic action of cysteine conjugates of chloro- and fluoroalkenes. Chem Biol Interact. 1985 Jun;54(1):15–31. doi: 10.1016/s0009-2797(85)80149-6. [DOI] [PubMed] [Google Scholar]
  21. Gu W., Bhatia K., Magrath I. T., Dang C. V., Dalla-Favera R. Binding and suppression of the Myc transcriptional activation domain by p107. Science. 1994 Apr 8;264(5156):251–254. doi: 10.1126/science.8146655. [DOI] [PubMed] [Google Scholar]
  22. Gu W., Cechova K., Tassi V., Dalla-Favera R. Opposite regulation of gene transcription and cell proliferation by c-Myc and Max. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2935–2939. doi: 10.1073/pnas.90.7.2935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Halleck M. M., Holbrook N. J., Skinner J., Liu H., Stevens J. L. The molecular response to reductive stress in LLC-PK1 renal epithelial cells: coordinate transcriptional regulation of gadd153 and grp78 genes by thiols. Cell Stress Chaperones. 1997 Mar;2(1):31–40. doi: 10.1379/1466-1268(1997)002<0031:tmrtrs>2.3.co;2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hann S. R., Dixit M., Sears R. C., Sealy L. The alternatively initiated c-Myc proteins differentially regulate transcription through a noncanonical DNA-binding site. Genes Dev. 1994 Oct 15;8(20):2441–2452. doi: 10.1101/gad.8.20.2441. [DOI] [PubMed] [Google Scholar]
  25. Harrington E. A., Bennett M. R., Fanidi A., Evan G. I. c-Myc-induced apoptosis in fibroblasts is inhibited by specific cytokines. EMBO J. 1994 Jul 15;13(14):3286–3295. doi: 10.1002/j.1460-2075.1994.tb06630.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Hayden P. J., Stevens J. L. Cysteine conjugate toxicity, metabolism, and binding to macromolecules in isolated rat kidney mitochondria. Mol Pharmacol. 1990 Mar;37(3):468–476. [PubMed] [Google Scholar]
  27. Hermeking H., Eick D. Mediation of c-Myc-induced apoptosis by p53. Science. 1994 Sep 30;265(5181):2091–2093. doi: 10.1126/science.8091232. [DOI] [PubMed] [Google Scholar]
  28. Hockenbery D., Nuñez G., Milliman C., Schreiber R. D., Korsmeyer S. J. Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature. 1990 Nov 22;348(6299):334–336. doi: 10.1038/348334a0. [DOI] [PubMed] [Google Scholar]
  29. Hurlin P. J., Quéva C., Koskinen P. J., Steingrímsson E., Ayer D. E., Copeland N. G., Jenkins N. A., Eisenman R. N. Mad3 and Mad4: novel Max-interacting transcriptional repressors that suppress c-myc dependent transformation and are expressed during neural and epidermal differentiation. EMBO J. 1995 Nov 15;14(22):5646–5659. doi: 10.1002/j.1460-2075.1995.tb00252.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Jänicke R. U., Lee F. H., Porter A. G. Nuclear c-Myc plays an important role in the cytotoxicity of tumor necrosis factor alpha in tumor cells. Mol Cell Biol. 1994 Sep;14(9):5661–5670. doi: 10.1128/mcb.14.9.5661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kang Y., Cortina R., Perry R. R. Role of c-myc in tamoxifen-induced apoptosis estrogen-independent breast cancer cells. J Natl Cancer Inst. 1996 Mar 6;88(5):279–284. doi: 10.1093/jnci/88.5.279. [DOI] [PubMed] [Google Scholar]
  32. Kato G. J., Barrett J., Villa-Garcia M., Dang C. V. An amino-terminal c-myc domain required for neoplastic transformation activates transcription. Mol Cell Biol. 1990 Nov;10(11):5914–5920. doi: 10.1128/mcb.10.11.5914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kato G. J., Lee W. M., Chen L. L., Dang C. V. Max: functional domains and interaction with c-Myc. Genes Dev. 1992 Jan;6(1):81–92. doi: 10.1101/gad.6.1.81. [DOI] [PubMed] [Google Scholar]
  34. Klefstrom J., Västrik I., Saksela E., Valle J., Eilers M., Alitalo K. c-Myc induces cellular susceptibility to the cytotoxic action of TNF-alpha. EMBO J. 1994 Nov 15;13(22):5442–5450. doi: 10.1002/j.1460-2075.1994.tb06879.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Kretzner L., Blackwood E. M., Eisenman R. N. Myc and Max proteins possess distinct transcriptional activities. Nature. 1992 Oct 1;359(6394):426–429. doi: 10.1038/359426a0. [DOI] [PubMed] [Google Scholar]
  36. Littlewood T. D., Hancock D. C., Danielian P. S., Parker M. G., Evan G. I. A modified oestrogen receptor ligand-binding domain as an improved switch for the regulation of heterologous proteins. Nucleic Acids Res. 1995 May 25;23(10):1686–1690. doi: 10.1093/nar/23.10.1686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Lotem J., Sachs L. A mutant p53 antagonizes the deregulated c-myc-mediated enhancement of apoptosis and decrease in leukemogenicity. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9672–9676. doi: 10.1073/pnas.92.21.9672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Lotem J., Sachs L. Regulation by bcl-2, c-myc, and p53 of susceptibility to induction of apoptosis by heat shock and cancer chemotherapy compounds in differentiation-competent and -defective myeloid leukemic cells. Cell Growth Differ. 1993 Jan;4(1):41–47. [PubMed] [Google Scholar]
  39. Lowe S. W., Ruley H. E., Jacks T., Housman D. E. p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell. 1993 Sep 24;74(6):957–967. doi: 10.1016/0092-8674(93)90719-7. [DOI] [PubMed] [Google Scholar]
  40. Majno G., Joris I. Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol. 1995 Jan;146(1):3–15. [PMC free article] [PubMed] [Google Scholar]
  41. Maki A., Berezesky I. K., Fargnoli J., Holbrook N. J., Trump B. F. Role of [Ca2+]i in induction of c-fos, c-jun, and c-myc mRNA in rat PTE after oxidative stress. FASEB J. 1992 Feb 1;6(3):919–924. doi: 10.1096/fasebj.6.3.1740241. [DOI] [PubMed] [Google Scholar]
  42. Marcu K. B., Bossone S. A., Patel A. J. myc function and regulation. Annu Rev Biochem. 1992;61:809–860. doi: 10.1146/annurev.bi.61.070192.004113. [DOI] [PubMed] [Google Scholar]
  43. Matsuoka M., Call K. M. Cadmium-induced expression of immediate early genes in LLC-PK1 cells. Kidney Int. 1995 Aug;48(2):383–389. doi: 10.1038/ki.1995.306. [DOI] [PubMed] [Google Scholar]
  44. McCarthy N. J., Whyte M. K., Gilbert C. S., Evan G. I. Inhibition of Ced-3/ICE-related proteases does not prevent cell death induced by oncogenes, DNA damage, or the Bcl-2 homologue Bak. J Cell Biol. 1997 Jan 13;136(1):215–227. doi: 10.1083/jcb.136.1.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Nuñez G., London L., Hockenbery D., Alexander M., McKearn J. P., Korsmeyer S. J. Deregulated Bcl-2 gene expression selectively prolongs survival of growth factor-deprived hemopoietic cell lines. J Immunol. 1990 May 1;144(9):3602–3610. [PubMed] [Google Scholar]
  46. Packham G., Ashmun R. A., Cleveland J. L. Cytokines suppress apoptosis independent of increases in reactive oxygen levels. J Immunol. 1996 Apr 15;156(8):2792–2800. [PubMed] [Google Scholar]
  47. Packham G., Cleveland J. L. Ornithine decarboxylase is a mediator of c-Myc-induced apoptosis. Mol Cell Biol. 1994 Sep;14(9):5741–5747. doi: 10.1128/mcb.14.9.5741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Packham G., Cleveland J. L. c-Myc and apoptosis. Biochim Biophys Acta. 1995 Jul 28;1242(1):11–28. doi: 10.1016/0304-419x(94)00015-t. [DOI] [PubMed] [Google Scholar]
  49. Packham G., Porter C. W., Cleveland J. L. c-Myc induces apoptosis and cell cycle progression by separable, yet overlapping, pathways. Oncogene. 1996 Aug 1;13(3):461–469. [PubMed] [Google Scholar]
  50. Peña A., Reddy C. D., Wu S., Hickok N. J., Reddy E. P., Yumet G., Soprano D. R., Soprano K. J. Regulation of human ornithine decarboxylase expression by the c-Myc.Max protein complex. J Biol Chem. 1993 Dec 25;268(36):27277–27285. [PubMed] [Google Scholar]
  51. Reisman D., Elkind N. B., Roy B., Beamon J., Rotter V. c-Myc trans-activates the p53 promoter through a required downstream CACGTG motif. Cell Growth Differ. 1993 Feb;4(2):57–65. [PubMed] [Google Scholar]
  52. Sakamuro D., Eviner V., Elliott K. J., Showe L., White E., Prendergast G. C. c-Myc induces apoptosis in epithelial cells by both p53-dependent and p53-independent mechanisms. Oncogene. 1995 Dec 7;11(11):2411–2418. [PubMed] [Google Scholar]
  53. Schreiber-Agus N., Chin L., Chen K., Torres R., Rao G., Guida P., Skoultchi A. I., DePinho R. A. An amino-terminal domain of Mxi1 mediates anti-Myc oncogenic activity and interacts with a homolog of the yeast transcriptional repressor SIN3. Cell. 1995 Mar 10;80(5):777–786. doi: 10.1016/0092-8674(95)90356-9. [DOI] [PubMed] [Google Scholar]
  54. Sentman C. L., Shutter J. R., Hockenbery D., Kanagawa O., Korsmeyer S. J. bcl-2 inhibits multiple forms of apoptosis but not negative selection in thymocytes. Cell. 1991 Nov 29;67(5):879–888. doi: 10.1016/0092-8674(91)90361-2. [DOI] [PubMed] [Google Scholar]
  55. Shi Y., Glynn J. M., Guilbert L. J., Cotter T. G., Bissonnette R. P., Green D. R. Role for c-myc in activation-induced apoptotic cell death in T cell hybridomas. Science. 1992 Jul 10;257(5067):212–214. doi: 10.1126/science.1378649. [DOI] [PubMed] [Google Scholar]
  56. Shibanuma M., Kuroki T., Nose K. Stimulation by hydrogen peroxide of DNA synthesis, competence family gene expression and phosphorylation of a specific protein in quiescent Balb/3T3 cells. Oncogene. 1990 Jul;5(7):1025–1032. [PubMed] [Google Scholar]
  57. Stevens J., Hayden P., Taylor G. The role of glutathione conjugate metabolism and cysteine conjugate beta-lyase in the mechanism of S-cysteine conjugate toxicity in LLC-PK1 cells. J Biol Chem. 1986 Mar 5;261(7):3325–3332. [PubMed] [Google Scholar]
  58. Tang N., Enger M. D. Cd(2+)-induced c-myc mRNA accumulation in NRK-49F cells is blocked by the protein kinase inhibitor H7 but not by HA1004, indicating that protein kinase C is a mediator of the response. Toxicology. 1993 Jul 28;81(2):155–164. doi: 10.1016/0300-483x(93)90007-f. [DOI] [PubMed] [Google Scholar]
  59. Tsujimoto Y. Stress-resistance conferred by high level of bcl-2 alpha protein in human B lymphoblastoid cell. Oncogene. 1989 Nov;4(11):1331–1336. [PubMed] [Google Scholar]
  60. Vamvakas S., Köster U. The nephrotoxin dichlorovinylcysteine induces expression of the protooncogenes c-fos and c-myc in LLC-PK1 cells--a comparative investigation with growth factors and 12-O-tetradecanoylphorbolacetate. Cell Biol Toxicol. 1993 Jan-Mar;9(1):1–13. doi: 10.1007/BF00755136. [DOI] [PubMed] [Google Scholar]
  61. Van de Water B., Kruidering M., Nagelkerke J. F. F-actin disorganization in apoptotic cell death of cultured rat renal proximal tubular cells. Am J Physiol. 1996 Apr;270(4 Pt 2):F593–F603. doi: 10.1152/ajprenal.1996.270.4.F593. [DOI] [PubMed] [Google Scholar]
  62. Vaux D. L., Cory S., Adams J. M. Bcl-2 gene promotes haemopoietic cell survival and cooperates with c-myc to immortalize pre-B cells. Nature. 1988 Sep 29;335(6189):440–442. doi: 10.1038/335440a0. [DOI] [PubMed] [Google Scholar]
  63. Wagner A. J., Kokontis J. M., Hay N. Myc-mediated apoptosis requires wild-type p53 in a manner independent of cell cycle arrest and the ability of p53 to induce p21waf1/cip1. Genes Dev. 1994 Dec 1;8(23):2817–2830. doi: 10.1101/gad.8.23.2817. [DOI] [PubMed] [Google Scholar]
  64. Wagner A. J., Meyers C., Laimins L. A., Hay N. c-Myc induces the expression and activity of ornithine decarboxylase. Cell Growth Differ. 1993 Nov;4(11):879–883. [PubMed] [Google Scholar]
  65. Wagner A. J., Small M. B., Hay N. Myc-mediated apoptosis is blocked by ectopic expression of Bcl-2. Mol Cell Biol. 1993 Apr;13(4):2432–2440. doi: 10.1128/mcb.13.4.2432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Weiss J. M., Lembach K. J., Boucek R. J., Jr A comparison of ornithine decarboxylases from normal and SV40-transformed 3T3 mouse fibroblasts. Biochem J. 1981 Jan 15;194(1):229–239. doi: 10.1042/bj1940229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Yu K., Chen Q., Liu H., Zhan Y., Stevens J. L. Signalling the molecular stress response to nephrotoxic and mutagenic cysteine conjugates: differential roles for protein synthesis and calcium in the induction of c-fos and c-myc mRNA in LLC-PK1 cells. J Cell Physiol. 1994 Nov;161(2):303–311. doi: 10.1002/jcp.1041610215. [DOI] [PubMed] [Google Scholar]
  68. Zervos A. S., Gyuris J., Brent R. Mxi1, a protein that specifically interacts with Max to bind Myc-Max recognition sites. Cell. 1993 Jan 29;72(2):223–232. doi: 10.1016/0092-8674(93)90662-a. [DOI] [PubMed] [Google Scholar]
  69. van de Water B., Zoeteweij J. P., de Bont H. J., Mulder G. J., Nagelkerke J. F. Role of mitochondrial Ca2+ in the oxidative stress-induced dissipation of the mitochondrial membrane potential. Studies in isolated proximal tubular cells using the nephrotoxin 1,2-dichlorovinyl-L-cysteine. J Biol Chem. 1994 May 20;269(20):14546–14552. [PubMed] [Google Scholar]

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