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:

Restoration of p53 expression sensitizes human papillomavirus type 16 immortalized human keratinocytes to CD95-mediated apoptosis

Oncogene volume 21, pages 165–175 (2002)Cite this article

Abstract

To understand the function of the individual oncogenes of HPV16 in modulating the cellular response to apoptogenic signals, we used human keratinocytes immortalized with either E6, E7 or E6/E7 oncoproteins as model system. Applying CD95 antibodies or recombinant CD95 ligand, only the E7-immortalized cells underwent extensive apoptosis. In contrast, E6- and E6/E7-expressing keratinocytes were resistant. Dominance of E6 correlated with significant down-regulation of p53, c-Myc, p21 and Bcl-2. CD95 was found to be reduced in resistant HPV-positive cells, while there were no quantitative differences in expression levels of FADD, FLICE/caspase-8 or caspase-3. Notably, in contrast to primary human keratinocytes, all immortalized cells showed a general reduction of c-FLIP, an inhibitory protein which normally prevents unscheduled CD95-induced apoptosis. E6- and E6/E7-positive keratinocytes, however, can be sensitized to CD95 apoptosis by blocking proteasome-mediated proteolysis. CD95-resistant HPV-positive cells underwent apoptosis within 3–5 h upon co-incubation with MG132 and agonistic antibodies or CD95 ligand, which was preceded by a strong re-expression of p53 and c-Myc, but not of other half-life controlled proteins such as Bax or IκBα. Blockage of proteasomal activity alone did not result in apoptosis, although the same set of pro-apoptotic proteins was up-regulated. Performing similar experiments with cervical carcinoma cells expressing mutated p53 (C33a) or with p53-‘null’ lung carcinoma cells (H1299), no CD95 cell killing occurred eventhough c-Myc was strong induced. These data indicate that the reduced bioavailability of p53 is a key-regulatory event in perturbation of CD95 signaling in HPV16 immortalized keratinocytes.

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
Figure 7

Similar content being viewed by others

References

  • Aguilar-Lemarroy A, Kirchhoff S, Whitaker N, Gariglio P, zur Hausen H, Krammer P, Rösl F . 2001 Int. J. Cancer 93: 823–831

  • Barbosa MS, Schlegel R . 1989 Oncogene 4: 1529–1532

  • Behrmann I, Walczak H, Krammer PH . 1994 Eur. J. Immunol. 24: 3057–3062

  • Bennett M, Macdonald K, Chan SW, Luzio JP, Simari R, Weissberg P . 1998 Science 282: 290–293

  • Boyer SN, Wazer DE, Band V . 1996 Cancer Res. 56: 4620–4624

  • Bristow RG, Jang A, Peacock J, Chung S, Benchimol S, Hill RP . 1994 Oncogene 9: 1527–1536

  • Brown J, Higo H, McKalip A, Herman B . 1997 J. Cell Biochem. 66: 245–255

  • Butz K, Geisen C, Ullmann A, Spitkovsky D, Hoppe-Seyler F . 1996 Int. J. Cancer 68: 506–513

  • Chaturvedi V, Qin J-Z, Denning MF, Choubey D, Diaz MO, Nickoloff BJ . 1999 J. Biol. Chem. 274: 23358–23367

  • Chéne P . 1998 J. Mol. Biol. 281: 205–209

  • Chellappan S, Kraus VB, Kroger B, Münger K, Howley PM, Phelps WC, Nevins JR . 1992 Proc. Natl. Acad. Sci. USA 89: 4549–4553

  • Chinnaiyan AM, O'Rourke K, Tewari M, Dixit VM . 1995 Cell 81: 505–512

  • Deng C, Zhang P, Harper JW, Elledge SJ, Leder P . 1995 Cell 82: 675–684

  • Dhein J, Daniel PT, Trauth BC, Oehm A, Moller P, Krammer PH . 1992 J. Immunol. 149: 3166–3173

  • Doorbar J, Parton A, Hartley K, Banks L, Crook T, Stanley M, Crawford L . 1990 Virology 178: 254–262

  • Drexler HC . 1997 Proc. Natl. Acad. Sci. USA 94: 855–860

  • Duerksen-Hughes PJ, Yang J, Schwartz SB . 1999 Virology 264: 55–65

  • El Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B . 1993 Cell 75: 817–825

  • El Deiry WS, Harper JW, O'Connor PM, Velculescu VE, Canman CE, Jackman J, Pietenpol JA, Burrell M, Hill DE, Wang Y . 1994 Cancer Res. 54: 1169–1174

  • Evan G, Littlewood T . 1998 Science 281: 1317–1322

  • Ferrari D, Stepczynska A, Los M, Wesselborg S, Schulze-Osthoff K . 1998 J. Exp. Med. 188: 979–984

  • Fukazawa T, Fujiwara T, Morimoto Y, Shao J, Nishizaki M, Kadowaki Y, Hizuta A, Owen-Schaub LB, Roth JA, Tanaka N . 1999 Oncogene 18: 2189–2199

  • Gehring S, Rottmann S, Menkel AR, Mertsching J, Krippner-Heidenreich A, Lüscher B . 2000 J. Biol. Chem. 275: 10413–10420

  • Griffiths DJ, Venables PJ, Weiss RA, Boyd MT . 1997 J. Virol. 71: 2866–2872

  • Gross-Mesilaty S, Reinstein E, Bercovich B, Tobias KE, Schwartz AL, Kahana C, Ciechanover A . 1998 Proc. Natl. Acad. Sci. USA 95: 8058–8063

  • Haas-Kogan DA, Kogan SC, Levi D, Dazin P, T'Ang A, Fung YK, Israel MA . 1995 EMBO J. 14: 461–472

  • Halbert CL, Demers GW, Galloway DA . 1991 J. Virol. 65: 473–478

  • Hartwell L . 1992 Cell 71: 543–546

  • Hawley-Nelson P, Vousden KH, Hubbert NL, Lowy DR, Schiller JT . 1989 EMBO J. 8: 3905–3910

  • Hengstermann A, Whitaker NJ, Zimmer D, Zentgraf H, Scheffner M . 1998 Oncogene 17: 2933–2941

  • Iftner T, Oft M, Bohm S, Wilczynski SP, Pfister H . 1992 J. Virol. 66: 4639–4646

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

  • Itoh N, Yonehara S, Ishii A, Yonehara M, Mizushima S, Sameshima M, Hase A, Seto Y, Nagata S . 1991 Cell 66: 233–243

  • Jacobson MD, Weil M, Raff MC . 1997 Cell 88: 347–354

  • Kaufmann SH, Desnoyers S, Ottaviano Y, Davidson NE, Poirier GG . 1993 Cancer Res. 53: 3976–3985

  • Khandjian EW, Meric C . 1986 Anal. Biochem. 159: 227–232

  • Kim CY, Tsai MH, Osmanian C, Graeber TG, Lee JE, Giffard RG, DiPaolo JA, Peehl DM, Giaccia AJ . 1997 Cancer Res. 57: 4200–4204

  • Kischkel FC, Hellbardt S, Behrmann I, Germer M, Pawlita M, Krammer PH, Peter ME . 1995 EMBO J. 14: 5579–5588

  • Knudson CM, Tung KS, Tourtellotte WG, Brown GA, Korsmeyer SJ . 1995 Science 270: 96–99

  • Kobayashi T, Ruan S, Jabbur JR, Consoli U, Clodi K, Shiku H, Owen-Schaub LB, Andreeff M, Reed JC, Zhang W . 1998 Cell Death. Differ. 5: 584–591

  • Korsmeyer SJ . 1999 Cancer Res. 59: 1693–1700

  • Langone JJ . 1982 J. Immunol. Methods 51: 3–22

  • Lee DH, Goldberg AL . 1998 Trends Cell Biol. 8: 397–403

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

  • Li B, Dou QP . 2000 Proc. Natl. Acad. Sci. USA 97: 3850–3855

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

  • Liang XH, Volkmann M, Klein R, Herman B, Lockett SJ . 1993 Oncogene 8: 2645–2652

  • Maki CG, Huibregtse JM, Howley PM . 1996 Cancer Res. 56: 2649–2654

  • Martin SJ, Lennon SV, Bonham AM, Cotter TG . 1990 J. Immunol. 145: 1859–1867

  • Meriin AB, Gabai VL, Yaglom J, Shifrin VI, Sherman MY . 1998 J. Biol. Chem. 273: 6373–6379

  • Mitsudomi T, Steinberg S, Nau M, Carbone D, DÁmico D, Bodner S, Oie H, Linnoila R, Mulshine J, Minna J, Gazdar A . 1992 Oncogene 7: 171–180

  • Miyashita T, Krajewski S, Krajewska M, Wang HG, Lin HK, Liebermann DA, Hoffman B, Reed JC . 1994 Oncogene 9: 1799–1805

  • Muzio M, Chinnaiyan AM, Kischkel FC, O'Rourke K, Shevchenko A, Ni J, Scaffidi C, Bretz JD, Zhang M, Gentz R, Mann M, Krammer PH, Peter ME, Dixit VM . 1996 Cell 85: 817–827

  • Müller M, Wilder S, Bannasch D, Israeli D, Lehlbach K, Li-Weber M, Friedman SL, Galle PR, Stremmel W, Oren M, Krammer PH . 1998 J. Exp. Med. 188: 2033–2045

  • Nair P, Nair KM, Jayaprakash PG, Pillai MR . 1999 Pathol. Oncol. Res. 5: 95–103

  • Natoli G, Ianni A, Costanzo A, De Petrillo G, Ilari I, Chirillo P, Balsano C, Levrero M . 1995 Oncogene 11: 1157–1164

  • Oda K, Arakawa H, Tanaka T, Matsuda K, Tanikawa C, Mori T, Nishimori H, Tamai K, Tokino T, Nakamura Y, Taya Y . 2000 Cell 102: 849–862

  • Oehm A, Behrmann I, Falk W, Pawlita M, Maier G, Klas C, Li-Weber M, Richards S, Dhein J, Trauth BC, Ponsting H, Krammer PH . 1992 J. Biol. Chem. 267: 10709–10715

  • Oltvai ZN, Milliman CL, Korsmeyer SJ . 1993 Cell 74: 609–619

  • Owen-Schaub LB, Zhang W, Cusack JC, Angelo LS, Santee SM, Fujiwara T, Roth JA, Deisseroth AB, Zhang WW, Kruzel E . 1995 Mol. Cell Biol. 15: 3032–3040

  • Pim D, Banks L . 1999 Oncogene 18: 7403–7408

  • Prendergast GC . 1999 Oncogene 18: 2967–2987

  • Reed JC . 1998 Oncogene 17: 3225–3236

  • Rolfe M, Chiu MI, Pagano M . 1997 J. Mol. Med. 75: 5–17

  • 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, Medema JP, Krammer PH, Peter ME . 1997 J. Biol. Chem. 272: 26953–26958

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

  • Scheffner M, Huibregtse JM, Howley PM . 1994 Proc. Natl. Acad. Sci. USA 91: 8797–8801

  • Scheffner M, Huibregtse JM, Vierstra RD, Howley PM . 1993 Cell 75: 495–505

  • Scheffner M, Münger K, Byrne JC, Howley PM . 1991 Proc. Natl. Acad. Sci. USA 88: 5523–5527

  • Scheffner M, Werness BA, Huibregtse JM, Levine AJ, Howley PM . 1990 Cell 63: 1129–1136

  • Schmitz I, Walczak H, Krammer PH, Peter ME . 1999 Cell Death. Differ. 6: 821–822

  • Schulze-Osthoff K, Krammer PH, Droge W . 1994 EMBO J. 13: 4587–4596

  • Seewaldt VL, Mrozek K, Dietze EC, Parker M, Caldwell LE . 2001 Cancer Res. 61: 616–624

  • Sheets EE, Crum CP, Yeh J . 1996 Gynecol. Oncol. 63: 94–100

  • Stöppler H, Stöppler MC, Johnson E, Simbulan-Rosenthal CM, Smulson ME, Iyer S, Rosenthal DS, Schlegel R . 1998 Oncogene 17: 1207–1214

  • Suda T, Takahashi T, Golstein P, Nagata S . 1993 Cell 75: 1169–1178

  • Talis AL, Huibregtse JM, Howley PM . 1998 J. Biol. Chem. 273: 6439–6445

  • Thome M, Schneider P, Hofmann K, Fickenscher H, Meinl E, Neipel F, Mattmann C, Burns K, Bodmer JL, Schroter M, Scaffidi C, Krammer PH, Peter ME, Tschopp J . 1997 Nature 386: 517–521

  • Trauth BC, Klas C, Peters AM, Matzku S, Moller P, Falk W, Debatin KM, Krammer PH . 1989 Science 245: 301–305

  • von Reyher U, Strater J, Kittstein W, Gschwendt M, Krammer PH, Moller P . 1998 Cancer Res. 58: 526–534

  • Winoto A . 1997 Curr. Opin. Immunol. 9: 365–370

  • Xu C, Meikrantz W, Schlegel R, Sager R . 1995 Proc. Natl. Acad. Sci. USA 92: 7829–7833

  • Yee C, Krishnan-Hewlett I, Baker CC, Schlegel R, Howley PM . 1985 Am. J. Pathol. 119: 361–366

  • Zheng TS, Schlosser SF, Dao T, Hingorani R, Crispe IN, Boyer JL, Flavell RA . 1998 Proc. Natl. Acad. Sci. USA 95: 13618–13623

  • zur Hausen H . 2000 J. Natl. Cancer Inst. 92: 690–698

Download references

Acknowledgements

We thank the Deutsche Akademische Austauschdienst and CONACyT for supporting A Aguilar-Lemarroy. We are also grateful to Dr Petra Boukamp (Deutsches Krebsforschungszentrum, Heidelberg) for providing primary human keratinocytes and to Dr Denise Galloway (Fred Hutchinson Cancer Center, Seattle, U.S.A.) for the amphotropic retroviruses.

Author information

Authors and Affiliations

  1. Forschungsschwerpunkt Angewandte Tumorvirologie, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, Heidelberg, 69120, Germany

    Adriana Aguilar-Lemarroy, Harald zur Hausen & Frank Rösl

  2. Genetics and Molecular Biology Department, CINVESTAV, Instituto Politecnico Nacional, Mexico-City, Mexico

    Patricio Gariglio

  3. School of Biochemistry and Molecular Genetics, University of New South Wales, Sydney, 2052, Australia

    Noel J Whitaker

  4. Forschungsschwerpunkt Tumorimmunologie, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 280, Heidelberg, 69120, Germany

    Sören T Eichhorst & Peter H Krammer

Authors
  1. Adriana Aguilar-Lemarroy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Patricio Gariglio
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Noel J Whitaker
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sören T Eichhorst
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Harald zur Hausen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Peter H Krammer
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Frank Rösl
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Frank Rösl.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Aguilar-Lemarroy, A., Gariglio, P., Whitaker, N. et al. Restoration of p53 expression sensitizes human papillomavirus type 16 immortalized human keratinocytes to CD95-mediated apoptosis. Oncogene 21, 165–175 (2002). https://doi.org/10.1038/sj.onc.1204979

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1204979

Keywords

This article is cited by

Search

Advanced search

Quick links