Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Identification of p21 as a target of cycloheximide-mediated facilitation of CD95-mediated apoptosis in human malignant glioma cells

Oncogene volume 20, pages 4757–4767 (2001)Cite this article

Abstract

Human glioma cell lines differ in their requirement for the inhibition of protein synthesis to activate the CD95-dependent killing pathway. CD95 ligand (CD95L) induced mitochondrial cytochrome c release and processing of caspases 3, 7, 8 and 9 in LN-18 cells in the absence of an inhibitor of protein synthesis, cycloheximide (CHX). These biochemical changes were observed in LN-229 cells only in the presence of CHX. The viral caspase inhibitor, cytokine response modifier (crm)-A, inhibited mitochondrial cytochrome c release, caspase processing and cell death under all conditions. Ectopic expression of BCL-XL prevented processing of caspase 8 in LN-18 cells but not in LN-229 cells. Thus, caspase 8 activation is amplified through the release of cytochrome c in LN-18 cells but occurs mainly at the receptor in LN-229 cells. In contrast to BCL-2, BCL-XL, X-linked inhibitor-of-apoptosis protein (XIAP) and FLICE-inhibitory protein (FLIP), the levels of the cyclin-dependent kinase (CDK) inhibitor, p21Waf/Cip1, rapidly decreased in response to CHX. P21 antisense oligonucleotides promoted caspase activation and mitochondrial cytochrome c release and induced strong sensitization to CD95-mediated apoptosis. These data place potentiating effects of CHX (i) to the activation of caspase 8 at the receptor in LN-229 cells as well as (ii) to a down-stream target at least in LN-18 cells, but probably both cell lines, that may be identical with p21Waf/Cip1.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • Asada M, Yamada T, Ichijo H, Delia D, Miyazono K, Fukumuro K, Mizutani S . 1999 EMBO J. 18: 1223–1234

  • Ashkenazi A, Dixit VM . 1998 Science 281: 1305–1308

  • Bossy-Wetzel E, Green DR . 1999 J. Biol. Chem. 274: 17484–17490

  • Clem RJ, Cheng EHY, Karp C, Kirsch DG, Ueno K, Takahashi A, Kastan MB, Griffin DE, Earnshaw WC, Veliuona MA, Hardwick JM . 1998 Proc. Natl. Acad. Sci. USA 95: 554–559

  • Deveraux QL, Leo E, Stennicke HR, Welsh K, Salvesen GS, Reed JC . 1999 EMBO J. 18: 5242–5251

  • Deveraux QL, Takahashi R, Salvesen GS, Reed JC . 1997 Nature 388: 300–304

  • Fearnhead HO, Rodriguez J, Govek EE, Guo W, Kobayashi R, Hannon G, Lazebnik YA . 1998 Proc. Natl. Acad. Sci. USA 95: 13664–13669

  • Fujita N, Nagahashi A, Nagashima K, Rokudai S, Tsuruo T . 1998 Oncogene 17: 1295–1304

  • Fulda S, Meyer E, Debatin KM . 2000 Cancer Res. 60: 3947–3956

  • Gervais JLM, Seth P, Zhang H . 1998 J. Biol. Chem. 273: 19207–19212

  • Glaser T, Wagenknecht B, Groscurth P, Krammer PH, Weller M . 1999 Oncogene 18: 5044–5053

  • Gorospe M, Wang X, Guyton KZ, Holbrook NJ . 1996 Mol. Cell Biol. 16: 6654–6660

  • Granville DJ, Shaw JR, Leong S, Carthy CM, Margaron P, Hunt DW, McManus BM . 1999 Am. J. Pathol. 155: 1021–1025

  • Griffith TS, Chin WA, Jackson GC, Lynch DH, Kubin MZ . 1998 J. Immunol. 161: 2833–2840

  • Gross A, Yin XM, Wang K, Wei MC, Jockel J, Milliman C, Erdjument-Bromage H, Tempst P, Korsmeyer SJ . 1999 J. Biol. Chem. 274: 1156–1163

  • Hueber A, Durka S, Weller M . 1998 FEBS Lett. 432: 155–157

  • Imai Y, Kimura T, Murakami A, Yajima N, Sakamaki K, Yonehara S . 1999 Nature 398: 777–785

  • Irmler M, Thome M, Hahne M, Schneider P, Hofmann K, Steiner V, Bodmer JL, Schröter M, Burns K, Mattmann C, Rimoldi D, French LE, Tschopp J . 1997 Nature 388: 190–195

  • Kirsch DG, Doseff A, Chau BN, Lim DS, De Souza-Pinto NC, Hansford R, Kastan MB, Lazebnik YA, Hardwick JM . 1999 J. Biol. Chem. 274: 21155–21161

  • Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD . 1997 Science 275: 1132–1136

  • Kondo S, Shinomura Y, Kanayama S, Higashimoto Y, Kiyohara T, Zushi S, Kitamura S, Ueyama H, Matsuzawa Y . 1998 Oncogene 17: 2585–2591

  • Krajewski S, Krajewska M, Shabaik A, Wang HG, Irie S, Fong L, Reed JC . 1994 Cancer Res. 54: 5501–5507

  • Kuwana T, Smith JJ, Muzio M, Dixit V, Newmeyer DD, Kornbluth S . 1998 J. Biol. Chem. 273: 16589–16594

  • Leverkus M, Neumann M, Mengling T, Rauch CT, Brocker EB, Krammer PH, Walczak H . 2000 Cancer Res. 60: 553–559

  • Levkau B, Koyama H, Raines EW, Clurman BE, Herren B, Orth K, Roberts JM, Ross R . 1998 Mol. Cell. 1: 553–563

  • Li H, Zhu H, Xu C, Yuan J . 1998 Cell 94: 491–501

  • Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES, Wang X . 1997 Cell 91: 479–489

  • Luo X, Budihardjo I, Zou H, Slaughter C, Wang X . 1998 Cell 94: 481–490

  • Martin SJ, Amarante-Mendes GP, Shi L, Chuang TH, Casiano CA, O'Brien GA, Fitzgerald P, Tan EM, Bokoch GM, Greenberg AH, Green DR . 1996 EMBO J. 15: 2407–2416

  • Naumann U, Durka S, Weller M . 1998 Oncogene 17: 1567–1575

  • Ng FW, Nguyen M, Kwan T, Branton PE, Nicholson DW, Cromlish JA, Shore GC . 1997 J. Cell Biol. 139: 327–338

  • Park JA, Kim KW, Kim SI, Lee SK . 1998 Eur. J. Biochem. 257: 242–248

  • Poluha W, Poluha DK, Chang B, Crosbie NE, Schonhoff CM, Kilpatrick DL, Ross AH . 1996 Mol. Cell Biol. 16: 1335–1341

  • Rieger J, Naumann U, Glaser T, Ashkenazi A, Weller M . 1998 FEBS Lett. 427: 124–128

  • Ruan S, Okcu MF, Ren JP, Chiao P, Andreeff M, Levin V, Zhang W . 1998 Cancer Res. 58: 1538–1543

  • Saleh A, Srinivasula SM, Acharya S, Fishel R, Alnemri ES . 1999 J. Biol. Chem. 274: 17941–17945

  • Scaffidi C, Fulda S, Srinivasan A, Friesen C, Li F, Tomaselli KJ, Debatin KM, Krammer PH, Peter ME . 1998 EMBO J. 17: 1675–1687

  • Scaffidi C, Schmitz I, Krammer PH, Peter ME . 1999 J. Biol. Chem. 274: 1541–1548

  • Schneider P, Holler N, Bodmer JL, Hahne M, Frei K, Fontana A, Tschopp J . 1998 J. Exp. Med. 187: 1205–1213

  • Slee EA, Harte MT, Kluck RM, Wolf BB, Casiano CA, Newmeyer DD, Wang HG, Reed JC, Nicholson DW, Alnemri ES, Green DR, Martin SJ . 1999 J. Cell Biol. 144: 281–292

  • Srinivasula SM, Ahmad M, Ottilie S, Bullrich F, Banks S, Wang Y, Fernandes-Alnemri T, Croce CM, Litwack G, Tomaselli KJ, Armstrong RC, Alnemri ES . 1997 J. Biol. Chem. 272: 18542–18545

  • Suzuki A, Tsutomi Y, Akahane K, Araki T, Miura M . 1998 Oncogene 17: 931–939

  • Suzuki A, Tsutomi Y, Miura M, Akahane K . 1999a Oncogene 18: 1239–1244

  • Suzuki A, Tsutomi Y, Yamamoto N, Shibutani T, Akahane K . 1999b Mol. Cell Biol. 19: 3842–3847

  • Suzuki A, Ito T, Kawano H, Hayashida M, Hayasaki Y, Tsutomi Y, Akahane K, Nakano T, Miura M, Shiraki K . 2000a Oncogene 19: 1346–1353

  • Suzuki A, Kawano H, Hayashida M, Hayasaki Y, Tsutomi Y, Akahane K . 2000b Cell Death Differ. 7: 721–728

  • Wagenknecht B, Schulz JB, Gulbins E, Weller M . 1998 Cell Death Differ. 5: 894–900

  • Wagenknecht B. Glaser T. Naumann U, Kügler S, Isenmann S, Bähr M, Korneluk R, Liston P, Weller M . 1999 Cell Death Differ. 6: 370–376

  • Wajant H, Haas E, Schwenzer R, Muhlenbeck F, Kreuz S, Schubert G, Grell M, Smith C, Scheurich P . 2000 J. Biol. Chem. 275: 24357–24366

  • Wang K, Yin XM, Chao DT, Milliman CL, Korsmeyer SJ . 1996 Genes Dev. 10: 2859–2869

  • Wang Z, Tuyle GV, Conrad D, Fisher PB, Dent P, Grant S . 1999 Cancer Res. 59: 1259–1267

  • Weller M, Frei K, Froscurth P, Krammer PH, Yonekawa Y, Fontana A . 1994 J. Clin. Invest. 94: 954–964

  • Weller M, Malipiero UV, Aguzzi A, Reed JC, Fontana A . 1995a J. Clin. Invest. 95: 2633–2643

  • Weller M, Malipiero UV, Rensing-Ehl A, Barr P, Fontana A . 1995b Cancer Res. 55: 2963–2944

  • Weller M, Rieger J, Grimmel C, Van Meir EG, De Tribolet N, Krajewski S, Reed JC, Von Deimling A, Dichgans J . 1998 Int. J. Cancer 79: 640–644

  • Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng TI, Jones DP, Wang X . 1997 Science 275: 1129–1132

  • Zhang Y, Fujita N, Tsuruo T . 1999a Oncogene 18: 1131–1138

  • Zhang Y, Fujita N, Tsuruo T . 1999b Int. J. Cancer 83: 790–797

  • Zou H, Li Y, Liu X, Wang X . 1999 J. Biol. Chem. 274: 11549–11556

Download references

Acknowledgements

Supported by the German Research Foundation (Deutsche Forschungsgemeinschaft, We 1502/3-2). T Glaser was supported by the BMBF (IZKF Tübingen). We thank PH Krammer for caspase 8 antibody, G Shore for Bap31 antibody, X Wang for BID antibody and Y Lazebnik for caspase 7 and 9 antibodies.

Author information

Authors and Affiliations

  1. Department of Neurology, Laboratory of Molecular Neuro-Oncology, University of Tübingen, Tübingen, Germany

    Tamara Glaser, Bettina Wagenknecht & Michael Weller

Authors
  1. Tamara Glaser
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bettina Wagenknecht
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael Weller
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Glaser, T., Wagenknecht, B. & Weller, M. Identification of p21 as a target of cycloheximide-mediated facilitation of CD95-mediated apoptosis in human malignant glioma cells. Oncogene 20, 4757–4767 (2001). https://doi.org/10.1038/sj.onc.1204498

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1204498

Keywords

This article is cited by

Search

Advanced search

Quick links