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Overcoming resistance to γ-rays in squamous carcinoma cells by poly-drug elevation of ceramide levels

Oncogene volume 23, pages 2703–2715 (2004)Cite this article

Abstract

Recent strategies to sensitize radioresistant tumours are based on combining γ-irradiation with inducers of apoptosis. We report that the combination of three inhibitors of sphingolipid metabolism, DL-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol.HCl(DL-PDMP)+imipramine±-D-erythro-2-(N-myristoylamino)-1-phenyl-1-propanol (D-MAPP), with 10-Gy irradiation triggers both mitotic and apoptotic killing in radioresistant SQ20B squamous carcinoma cells. In these cells, apoptosis is defective due to a lack of ceramide generation upstream, which cannot be explained by sphingomyelinase (neutral and acidic) deficiency or rapid derivation to the sphingolipid pathway. We present evidence of a functional transduction death pathway when ceramide generation is restored, which involves the mitochondrial-mediated pathway coupled to alterations in redox status and to executive caspases activation. The poly-drug treatment restored apoptosis to levels similar to those observed in radiosensitive SCC61 squamous carcinoma cells. Simultaneous exposure to γ-irradiation and poly-drug treatment acted synergistically in SQ20B cells to produce a marked increase in both mitochondrial dysfunction and caspase cleavage, which led to a 7.8-fold increase in apoptosis within 48 h, relative to irradiated cells. Moreover, the results suggest that the ceramide released by irradiation or poly-drug treatment converges upon common cellular targets. Modulation of endogenous ceramide levels by inhibitors of sphingolipid metabolism may represent a new cellular target for the sensitization of radioresistant tumours to γ-ray therapy.

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References

  • Alphonse G, Aloy MT, Broquet P, Gerard JP, Louisot P, Rousson R and Rodriguez-Lafrasse C . (2002). Int. J. Radiat. Biol., 78, 821–835.

  • Bidere N, Lorenzo HK, Carmona S, Laforge M, Harper F, Dumont C and Senik A . (2003). J. Biol. Chem., 278, 31401–31411.

  • Bielawska A, Greenberg MS, Perry D, Jayadev S, Shayman JA, McKay C and Hannun YA . (1996). J. Biol. Chem., 271, 12646–12654.

  • Birbes H, Bawab SE, Obeid LM and Hannun YA . (2002). Adv. Enzyme Regul., 42, 113–129.

  • Bruno AP, Laurent G, Averbeck D, Demur C, Bonnet J, Bettaieb A, Levade T and Jaffrezou JP . (1998). Cell Death Differ., 5, 172–182.

  • Chmura SJ, Mauceri HJ, Advani S, Heimann R, Beckett MA, Nodzenski E, Quintans J, Kufe DW and Weichselbaum RR . (1997a). Cancer Res., 57, 4340–4347.

  • Chmura SJ, Nodzenski E, Beckett MA, Kufe DW, Quintans J and Weichselbaum RR . (1997b). Cancer Res., 57, 1270–1275.

  • Choi MS, Anderson MA, Zhang Z, Zimonjic DB, Popescu N and Mukherjee AB . (2003). Gene, 315, 113–122.

  • Cuvillier O and Levade T . (2001). Blood, 98, 2828–2836.

  • Daniel PT . (2000). Leukemia, 14, 2035–2044.

  • Decaudin D, Marzo I, Brenner C and Kroemer G . (1998). Int. J. Oncol., 12, 141–152.

  • Di Bartolomeo S and Spinedi A . (2001). Biochem. Biophys. Res. Commun., 288, 269–274.

  • El-Assaad W, Kozhaya L, Araysi S, Panjarian S, Bitar FF, Baz E, El-Sabban ME and Dbaibo GS . (2003). Biochem. J., 376, 725–732.

  • Fiske CH and Subbarow Y . (1925). J. Biol. Chem., 66, 375–400.

  • Garcia-Ruiz C, Colell A, Mari M, Morales A and Fernandez-Checa JC . (1997). J. Biol. Chem., 272, 11369–11377.

  • Gewirtz DA . (2000). Breast Cancer Res. Treat., 62, 223–235.

  • Ghafourifar P, Klein SD, Schucht O, Schenk U, Pruschy M, Rocha S and Richter C . (1999). J. Biol. Chem., 274, 6080–6084.

  • Grazide S, Maestre N, Veldman RJ, Bezombes C, Maddens S, Levade T, Laurent G and Jaffrezou JP . (2002). FASEB J., 16, 1685–1687.

  • Gulbins E and Kolesnick R . (2003). Oncogene, 22, 7070–7077.

  • Ito A, Uehara T, Tokumitsu A, Okuma Y and Nomura Y . (1999). Biochim. Biophys. Acta, 1452, 263–274.

  • Itoh M, Kitano T, Watanabe M, Kondo T, Yabu T, Taguchi Y, Iwai K, Tashima M, Uchiyama T and Okazaki T . (2003). Clin. Cancer Res., 9, 415–423.

  • Jacobson K and Dietrich C . (1999). Trends Cell Biol., 9, 87–91.

  • Jaffrezou JP, Bruno AP, Moisand A, Levade T and Laurent G . (2001). FASEB J., 15, 123–133.

  • Kimura K, Markowski M, Edsall LC, Spiegel S and Gelmann EP . (2003). Cell Death Differ., 10, 240–248.

  • Kokunai T, Urui S, Tomita H and Tamaki N . (2001). J. Neurooncol., 51, 111–119.

  • Kolesnick R and Fuks Z . (2003). Oncogene, 22, 5897–5906.

  • Kurzchalia TV and Parton RG . (1999). Curr. Opin. Cell Biol., 11, 424–431.

  • Lavie Y, Cao H, Bursten SL, Giuliano AE and Cabot MC . (1996). J. Biol. Chem., 271, 19530–19536.

  • Lavrentiadou SN, Chan C, Kawcak T, Ravid T, Tsaba A, Rasooly R and Goldkorn T . (2001). Am. J. Respir. Cell Mol. Biol., 25, 676–684.

  • Lee JU, Hosotani R, Wada M, Doi R, Kosiba T, Fujimoto K, Miyamoto Y, Tsuji S, Nakajima S, Nishimura Y and Imamura M . (1999). Eur. J. Cancer, 35, 1374–1380.

  • Li W, Yuan X, Nordgren G, Dalen H, Dubowchik GM, Firestone RA and Brunk UT . (2000). FEBS Lett., 470, 35–39.

  • Lin J, Page C, Jin X, Sethi AO, Patel R and Nunez G . (2001). Anticancer Res., 21, 831–839.

  • Liu B and Hannun YA . (1997). J. Biol. Chem., 272, 16281–16287.

  • Liu B, Andrieu-Abadie N, Levade T, Zhang P, Obeid LM and Hannun YA . (1998). J. Biol. Chem., 273, 11313–11320.

  • Liu YY, Han TY, Giuliano AE, Ichikawa S, Hirabayashi Y and Cabot MC . (1999). Exp. Cell Res., 252, 464–470.

  • Liu YY, Han TY, Giuliano AE, Hansen N and Cabot MC . (2000). J. Biol. Chem., 275, 7138–7143.

  • Lozano J, Menendez S, Morales A, Ehleiter D, Liao WC, Wagman R, Haimovitz-Friedman A, Fuks Z and Kolesnick R . (2001). J. Biol. Chem., 276, 442–448.

  • Lucci A, Cho WI, Han TY, Giuliano AE, Morton DL and Cabot MC . (1998). Anticancer Res., 18, 475–480.

  • Matsuo N, Nomura T and Imokawa G . (1992). Biochim. Biophys. Acta, 1116, 97–103.

  • McIlwrath AJ, Vasey PA, Ross GM and Brown R . (1994). Cancer Res., 54, 3718–3722.

  • Mendez AJ, Lin G, Wade DP, Lawn RM and Oram JF . (2001). J. Biol. Chem., 276, 3158–3166.

  • Michael JM, Lavin MF and Watters DJ . (1997). Cancer Res., 57, 3600–3605.

  • Neuschwander-Tetri BA and Roll FJ . (1989). Anal. Biochem., 179, 236–241.

  • Pena LA, Fuks Z and Kolesnick RN . (2000). Cancer Res., 60, 321–327.

  • Perry DK . (1999). Biochem. Soc. Trans., 27, 399–404.

  • Petit PX, Zamzami N, Vayssiere JL, Mignotte B, Kroemer G and Castedo M . (1997). Mol. Cell. Biochem., 174, 185–188.

  • Radin NS . (2003). Biochem. J., 371, 243–256.

  • Raisova M, Goltz G, Bektas M, Bielawska A, Riebeling C, Hossini AM, Eberle J, Hannun YA, Orfanos CE and Geilen CC . (2002). FEBS Lett., 516, 47–52.

  • Ramstedt B and Slotte JP . (2002). FEBS Lett., 531, 33–37.

  • Riboni L, Campanella R, Bassi R, Villani R, Gaini SM, Martinelli-Boneschi F, Viani P and Tettamanti G . (2002). Glia, 39, 105–113.

  • Rodriguez-Lafrasse C, Rousson R, Pentchev PG, Louisot P and Vanier MT . (1994). Biochim. Biophys. Acta, 1226, 138–144.

  • Rodriguez-Lafrasse C, Alphonse G, Broquet P, Aloy MT, Louisot P and Rousson R . (2001). Biochem. J., 357, 407–416.

  • Rodriguez-Lafrasse C, Alphonse G, Aloy MT, Ardail D, Gerard JP, Louisot P and Rousson R . (2002). Int. J. Cancer, 101, 589–598.

  • Rosenwald AG and Pagano RE . (1994). J. Lipid Res., 35, 1232–1240.

  • Santana P, Pena LA, Haimovitz-Friedman A, Martin S, Green D, McLoughlin M, Cordon-Cardo C, Schuchman EH, Fuks Z and Kolesnick R . (1996). Cell, 86, 189–199.

  • Sautin Y, Takamura N, Shklyaev S, Nagayama Y, Ohtsuru A, Namba H and Yamashita S . (2000). Thyroid, 10, 733–740.

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

  • Selzner M, Bielawska A, Morse MA, Rudiger HA, Sindram D, Hannun YA and Clavien PA . (2001). Cancer Res., 61, 1233–1240.

  • Shi YQ, Wuergler FE, Blattmann H and Crompton NE . (2001). Radiat. Environ. Biophys., 40, 301–308.

  • Shinoura N, Sakurai S, Asai A, Kirino T and Hamada H . (2001). Int. J. Cancer, 93, 252–261.

  • Sietsma H, Veldman RJ, Kolk D, Ausema B, Nijhof W, Kamps W, Vellenga E and Kok JW . (2000). Clin. Cancer Res., 6, 942–948.

  • Solary E, Plenchette S, Sordet O, Rebe C, Ducoroy P, Filomenko R, Bruey JM, Droin N and Corcos L . (2001). Therapie, 56, 511–518.

  • Stoka V, Turk B, Schendel SL, Kim TH, Cirman T, Snipas SJ, Ellerby LM, Bredesen D, Freeze H, Abrahamson M, Bromme D, Krajewski S, Reed JC, Yin XM, Turk V and Salvesen GS . (2001). J. Biol. Chem., 276, 3149–3157.

  • Sugiki H, Hozumi Y, Maeshima H, Katagata Y, Mitsuhashi Y and Kondo S . (2000). Br. J. Dermatol., 143, 1154–1163.

  • Tepper AD, de Vries E, van Blitterswijk WJ and Borst J . (1999). J. Clin. Invest., 103, 971–978.

  • Tepper AD, Diks SH, van Blitterswijk WJ and Borst J . (2000). J. Biol. Chem., 275, 34810–33487.

  • Thomas RL, Matsko CM, Lotze MT and Amoscato AA . (1999). J. Biol. Chem., 274, 30580–30588.

  • Vanier MT, Revol A and Fichet M . (1980). Clin. Chim. Acta, 106, 257–267.

  • Veldman RJ, Mita A, Cuvillier O, Garcia V, Klappe K, Medin JA, Campbell JD, Carpentier S, Kok JW and Levade T . (2003). FASEB J., 17, 1144–1146.

  • Yokogawa K, Ishizaki J, Ohkuma S and Miyamoto K . (2002). Methods Find. Exp. Clin. Pharmacol., 24, 81–93.

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Acknowledgements

This work was supported by the Institut National de la Santé et de la Recherche Médicale and by the French ETOILE project (Espace de Traitement Oncologique par les Ions Légers). We thank Colette Berthet and all the technicians from the Biochemical Laboratory of the Cardiovascular and Pneumological Hospital (Lyon, France) for ceramide and glutathione quantification, and also the technicians from the haematology laboratory (Lyon-Sud Hospital Center) for their help with flow cytometry. We are grateful to the personnel of the Radiotherapy Department (under Dr P. Romestaing) for allowing us access to the irradiators of the Lyon-Sud Hospital Center.

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  1. Department of Biochemistry, INSERM U189, Lyon-Sud Medical School, BP12, Oullins Cedex, 69921, France

    Gersende Alphonse, Clara Bionda, Marie-Thérèse Aloy, Dominique Ardail, Robert Rousson & Claire Rodriguez-Lafrasse

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Correspondence to Claire Rodriguez-Lafrasse.

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Alphonse, G., Bionda, C., Aloy, MT. et al. Overcoming resistance to γ-rays in squamous carcinoma cells by poly-drug elevation of ceramide levels. Oncogene 23, 2703–2715 (2004). https://doi.org/10.1038/sj.onc.1207357

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