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Flavonoid quercetin sensitizes a CD95-resistant cell line to apoptosis by activating protein kinase Cα

Oncogene volume 22, pages 3330–3342 (2003)Cite this article

Abstract

We previously demonstrated that quercetin, a naturally occurring flavonoid with strong antioxidant properties, was able to enhance programmed cell death in HPB-acute lymphoblastic leukemia (ALL) cell line, derived from a human tymoma, when associated with the agonistic anti-CD95 monoclonal antibody. Here, we report that HPB-ALL cells are normally resistant to CD95-mediated apoptosis, and quercetin is able to sensitize this cell line through a mechanism independent of its antioxidant properties. In fact, other compounds structurally and functionally similar to quercetin, when associated with anti-CD95 antibody did not induce any CD95-mediated apoptosis, still maintaining their antioxidant capacity. We found that quercetin effects are mediated by the activation of PKCα. Treatment of HPB-ALL cells with quercetin slightly decreased PKCα activity, but when the flavonoid was associated with anti-CD95, the kinase activity increased by 12-fold with respect to the treatment with quercetin. In addition, overexpression of PKCα induced programmed cell death in the absence of any additional stimulus, while a kinase-defective mutant of PKCα was ineffective. Our data confirm the involvement of specific PKC isoforms in CD95 signaling and suggest, for the first time, that quercetin targets this pathway increasing apoptogenic response in a cell line resistant to CD95-mediated apoptosis.

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Abbreviations

DCFDA:

2′-7′-dichlorofluorescin diacetate

DMSO:

dimethylsulfoxide

IETD-AFC:

Ile-Glu-Thr-Asp-7-amino-4-trifluoromethyl coumarin

LB:

lysis buffer

LEHD-AFC:

Leu-Glu-Hys-Asp-7-amino-4-trifluoromethyl coumarin

PBS:

phosphate-buffered saline

PCD:

programmed cell death

PKC:

protein kinase C

PI3K:

inositol 1,4,5-triphosphate kinase

PMA:

12-O-tetradecanoylphorbol-13-acetate

T-TBS:

Tween-20/Tris-buffered saline

ROS:

reactive oxygen species

Z-DEVD-AFC:

carbobenzoxy-Asp-Glu-Val-Asp-7-amino-4-trifluoromethyl coumarin

References

  • Agullo G, Gamet-Payrastre L, Manenti S, Viala C, Remesey C, Chap H and Payrastre B . (1997). Biochem. Pharm., 53, 1649–1657.

  • Alderson MR, Tough TW, Braddy S, Davis-Smith T, Roux E, Schooley K, Miller RE and Lynch DH . (1994). Int. Immunol., 6, 1799–1806.

  • Avila M, Cansado J, Harter K, Velasco J and VN . (1996). Adv. Exp. Med. Biol., 401, 101–110.

  • Avila MA, Velasco JA, Cansado J and Notario V . (1994). Cancer Res., 54, 2424–2428.

  • Bors W, Heller W, Michel C and Saran M . (1990). Methods Enzymol., 186, 343–355.

  • Bradford MM . (1976). Anal. Biochem., 72, 248–254.

  • Caltagirone S, Rossi C, Poggi A, Ranelletti F, Natali P, Brunetti M, Aiello F and MP . (2000). Int. J. Cancer, 87, 595–600.

  • Casagrande F and Darbon J . (2001). Biochem. Pharmacol., 61, 1205–1215.

  • Chemoprevention Working Group. (1999). Cancer Res., 59, 4743–4758.

  • Chinnaiyan AM, O’Rourke K, Tewari M and Dixit VM . (1995). Cell, 81, 505–512.

  • Clement MV and Stamenkovic I . (1996). EMBO J., 15, 216–225.

  • Cuvillier O, Rosenthal DS, Smulson ME and Spiegel S . (1998). J. Biol. Chem., 273, 2910–2916.

  • Davies SP, Reddy H, Caivano M and Cohen P . (2000). Biochem. J., 351, 95–105.

  • Debatin KM and Krammer P . (1995). Leukemia, 9, 815–820.

  • Emoto Y, Manome Y, Meinhardt G, Kisaki H, Kharbanda S, Robertson M, Ghayur T, Wong WW, Kamen R, Weichselbaum R and Kufe D . (1995). EMBO J., 14, 6148–6156.

  • Fautz R, Husein B and Hechenberger C . (1991). Mutat. Res., 253, 173–179.

  • Ferrandina G, Almadori G, Maggiano N, Lanza P, Ferlini C, Cattani P, Piantelli M, Scambia G and Ranelletti FO . (1998). Int. J. Cancer, 77, 747–754.

  • Fiers W, Beyaert R, Declercq W and Vandenabeele P . (1999). Oncogene, 18, 7719–7730.

  • Frutos S, Moscat J and Diaz-Meco MT . (1999). J. Biol. Chem., 274, 10765–10770.

  • Galati G, Teng S, Moridani MY, Chan TS and O-Brien PJ . (2000). Drug Metabol. Drug Interact., 17, 311–349.

  • Gamet-Payrastre L, Manenti S, Gratacap MP, Tulliez J, Chap H and Payrastre B . (1999). Gen. Pharmacol., 32, 279–286.

  • Gillis S and Watson J . (1980). J. Exp. Med., 152, 1709–1719.

  • Goldberg D, Tsang E, Karumanchiri A, Diamandis E, Soleas G and Ngn E . (1996). Anal. Chem., 68, 1688–1694.

  • Gomez-Angelats M, Bortner CD and Cidlowski JA . (2000). J. Biol. Chem., 275, 19609–19619.

  • Gomez-Angelats M and Cidlowski JA . (2001). J. Biol. Chem., 276, 44944–44952.

  • Gschwendt M, Dieterich S, Rennecke J, Kittstein W, Mueller HJ and Johannes FJ . (1996). FEBS Lett., 392, 77–80.

  • Hagiwara M, Inoue S, Tanaka T, Nunoki K, Ito M and Hidaka H . (1988). Biochem. Pharmacol., 37, 2987–2992.

  • Hangartner M . (2000). Nature, 407, 770–776.

  • Hausler P, Papoff G, Eramo A, Reif K, Cantrell DA and Ruberti G . (1998). Eur. J. Immunol., 28, 57–69.

  • Ishikawa Y and Kitamura M . (2000). Biochem. Biophys. Res. Commun., 279, 629–634.

  • Iwao K and Tsukamoto I . (1999). Biochim. Biophys. Acta, 1427, 112–120.

  • Jacobson M, Weil M and Raff M . (1997). Cell, 88, 347–354.

  • Jamieson L, Carpenter L, Biden TJ and Fields AP . (1999). J. Biol. Chem., 274, 3927–3930.

  • Jarvis WD, Turner AJ, Povirk LF, Traylor RS and Grant S . (1994). Cancer Res., 54, 1707–1714.

  • Kang T and Liang N . (1997). Biochem. Pharmacol., 54, 1013–1018.

  • Kang ZC, Tsai SJ and Lee H . (1999). Nutr. Cancer, 35, 175–179.

  • Klas C, Debatin KM, Jonker RR and Krammer PH . (1993). Int. Immunol., 5, 625–630.

  • Krammer P . (2000). Nature, 407, 789–795.

  • Krammer PH, Galle PR, Moller P and Debatin KM . (1998). Advances in Cancer Research, Vol. 65. Vande Woude GF and Klein, G. (eds). Academic Press, Inc.: San Diego, pp. 251–272.

    Google Scholar 

  • Laemmli UK . (1970). Nature, 227, 680–685.

  • Larocca LM, Ranelletti FO, Maggiano N, Rutella S, La Barbera EO, Rumi C, Serra F, Voso MT, Piantelli M, Teofili L and Leone G . (1997). Int. J. Cancer, 73, 75–83.

  • Le Good J, Ziegler W, Parekh D, Alessi D, Cohen P and Parker P . (1998). Science, 281, 2042–2045.

  • Le Marchand L, Murphy S, Hankin J, Wilkens L and LN K . (2000). J. Natl. Cancer Inst., 92, 154–160.

  • Li W, Zhang J, Flechner L, Hyun T, Yam A, Franke TF and Pierce JH . (1999). Oncogene, 18, 6564–6572.

  • Lowe S and Lin A . (2000). Carcinogenesis, 21, 485–495.

  • Mahmoud N, Carothers A, Grunberger D, Bilinski R, Churchill M, Martucci C, Newmark H and Bertagnolli M . (2000). Carcinogenesis, 21, 921–927.

  • Mandil R, Ashkenazi E, Blass M, Kronfeld I, Kazimirsky G, Rosenthal G, Umansky F, Lorenzo P, Blumberg P and Brodie C . (2001). Cancer Res., 61, 4612–4619.

  • Middleton EJ and Kandaswami C . (1993). The Flavonoid: Advance Research Since 1986. Hairbone JH and Liss AR (eds): New York, pp. 619–652.

    Book  Google Scholar 

  • Mizuno K, Noda K, Araki T, Imaoka T, Kobayashi Y, Akita Y, Shimonoka M, Kishi S and Ohno S . (1997). Eur. J. Biochem., 250, 7–18.

  • Morikawa S, Tatsumi E, Baba M, Harada T and Yasuhira K . (1978). Int. J. Cancer, 21, 166–170.

  • Mouria M, Gukovskaya AS, Jung Y, Buechler P, Hines OJ, Reber H and Pandol SJ . (2002). Int. J. Cancer, 98, 761–769.

  • 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 and Dixit VM . (1996). Cell, 85, 817–827.

  • Nagata S . (1997). Cell, 88, 355–365.

  • Noroozi M, Burns J, Crozier A, Kelly IE and Lean ME . (2000). Eur. J. Clin. Nutr., 54, 143–149.

  • Parekh D, Ziegler W and Parker P . (2000). EMBO J., 19, 496–503.

  • Plaumann B, Fritsche M, Rimpler H, Brandner G and Hess RD . (1996). Oncogene, 13, 1605–1614.

  • Ranelletti F, Maggiano N, Serra F, Ricci R, Larocca L, Lanza P, Scambia G, Fattorossi A, Capelli A and Piantelli M . (2000). Int. J. Cancer, 85, 438–445.

  • Reddy BS . (2000). Cancer Epidemiol. Biomarkers Prev., 9, 239–247.

  • Rong Y, Yang EB, Zhang K and Mack P . (2000). Anticancer Res., 20, 4339–4345.

  • Ruiz-Ruiz MC, Robledo G, Font J, Izquierdo M and Lopez-Rivas A . (1999). J. Immunol., 163, 4737–4746.

  • Russo M, Palumbo R, Tedesco I, Mazzarella G, Russo P, Iacomino G and Russo G . (1999). FEBS Lett., 462, 322–328.

  • 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.

  • Scaffidi C, Schmitz I, Korsmeyer S, Krammer PH and Peter ME . (1999). J. Biol. Chem., 274, 22532–22538.

  • Shao RG, Cao CX and Pommier Y . (1997). J. Biol. Chem., 272, 31321–31325.

  • Shimizu T, Cao CX, Shao RG and Pommier Y . (1998). J. Biol. Chem., 273, 8669–8674.

  • Stavric B . (1994). Clinical Biochem., 27, 319–332.

  • Toullec D, Pianetti P, Coste H, Bellevergue P, Grand-Perret T, Ajakane M, Baudet V, Boissin P, Boursier E and Loriolle F . (1991). J. Biol. Chem., 266, 15771–15781.

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

  • Walker E, Pacold M, Perisic O, Stephens L, Hawkins P, Wymann M and Williams R . (2000). Mol. Cell, 6, 909–919.

  • Wang I, Lin-Shiau S and Lin J . (1999). Eur. J. Cancer, 35, 1517–1525.

  • Weber G, Shen F, Yang H, Prajda N and Li W . (1999). Adv. Enzyme Regul., 39, 51–66.

  • Weisburger J . (2000). Am. J. Clin. Nutr., 71, 1710S–1714S.

  • Yang C, Landau J, Huang M and Newmark H . (2001). Annu. Rev. Nutr., 21, 381–406.

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Acknowledgements

We thank the following colleagues for providing us several reagents used in the present study: Dr Weiqun Li for PKCα expressing vector; Drs Jae-Won Soh and I Bernard Weinstein for pHACE-PKC-KR plasmid; Dr Mauro Rossi for Jurkat cell line; Dr Luigia Santella for GF109203X. We also thank Drs Giavincenzo Barba and Paola Russo for their help in statistical analysis. This work has been supported by a grant from Istituto Superiore di Sanità (Roma, Italy), research project: ‘Natural antioxidants and effects on chronic-degenerative diseases.’

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  1. Istituto di Scienze dell’Alimentazione, Consiglio Nazionale delle Ricerche, Avellino, 83100, Italy

    Maria Russo, Rosanna Palumbo, Annalisa Mupo, Giuseppe Iacomino, Annamaria Scognamiglio, Idolo Tedesco, Giovanni Galano & Gian Luigi Russo

  2. Stazione Zoologica ‘Anton Dohrn’, Napoli, 80121, Italy

    Mariarosaria Tosto

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Correspondence to Gian Luigi Russo.

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Russo, M., Palumbo, R., Mupo, A. et al. Flavonoid quercetin sensitizes a CD95-resistant cell line to apoptosis by activating protein kinase Cα. Oncogene 22, 3330–3342 (2003). https://doi.org/10.1038/sj.onc.1206493

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