Abstract
Overexpression of EGF receptors and constitutive cyclin D1 expression are frequently associated with human squamous carcinomas. We have now investigated whether these parameters influence susceptibility to okadaic acid induced cell death in EGF-receptor overexpressing mutant p53 A431 human carcinoma. Exposure of these cells to 20 nM okadaic acid induced apoptosis-associated caspase 3 activation, DNA fragmentation, cleavage of Poly ADP-Ribose Polymerase (PARP), p53-independent expression of pro-apoptotic bax, and loss of proliferation-promoting cyclin D1. All these alterations were antagonized by concurrent addition of exogenous EGF. Ectopic overexpression of the cyclin D1 gene in A431 carcinoma conferred resistance to 20 nM okadaic acid irrespective of exogenous EGF, associated with a parallel induction of anti-apoptotic bcl-2. Treatment with a subtoxic concentration of a bispecific bcl-2/bcl xL antisense oligonucleotide cooperated with okadaic acid to down-regulate bcl-2 and sensitize cyclin D1-overexpressing cells to okadaic acid. Although EGF protects EGF-R proficient epithelial cells from diverse apoptotic stimuli through Mcl-1, this is the first report demonstrating that cyclin D1 overexpression provides an EGF independent protection from okadaic acid-induced cell death through induction of bcl-2. We also show that this anti-apoptotic effect of cyclin D1 overexpression, can be partly antagonized with antisense strategies that down-regulate anti-apoptotic bcl-2 family members.
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Abbreviations
- EGF-R:
-
epidermal growth factor receptor
- EGF:
-
epidermal growth factor
- PARP:
-
Poly ADP-Ribose Polymerase
References
Hall M, Peters G. Genetic alterations of cyclins, cyclin-dependent kinases, and Cdk inhibitors in human cancer. Adv Cancer Res 1996; 68: 67–108.
Han EK, Lim JT, Arber N, Rubin MA, Xing WQ, Weinstein IB. Cyclin D1 expression in human Prostate carcinoma cell lines and primary tumors. Prostate 1998; 35(2): 95–101.
Perry JE, Grossmann ME, Tindall DJ. Epidermal growth factor induces cyclin D1 in a human Prostate cancer cell line. Prostate 1998; 35(2): 117–124.
Poch B, Gansauge F, Schwarz A, et al. Epidermal growth factor induces cyclin D1 in human pancreatic carcinoma: evidence for a cyclin D1-dependent cell cycle progression. Pancreas 2001; 23(3): 280–287.
Kornmann M, Ishiwata T, Itakura J, Tangvoranuntakul P, Beger HG, Korc M. Increased cyclin D1 in human pancreatic cancer is associated with decreased postoperative survival. Oncology 1998; 55(4): 363–369.
Chen Y, Martinez LA, LaCava M, Coghlan L, Conti CJ. Increased cell growth and tumorigenicity in human prostate LNCaP cells by overexpression to cyclin D1. Oncogene 1998; 16(15): 1913–1920.
Saikawa Y, Kubota T, Otani Y, Kitajima M, Modlin IM. Cyclin D1 antisense oligonucleotide inhibits cell growth stimulated by epidermal growth factor and induces apoptosis of gastric cancer cells. Jpn J Cancer Res 2001; 92(10): 1102–1109.
Kornmann M, Arber N, Korc M. Inhibition of basal and mitogen-stimulated pancreatic cancer cell growth by cyclin D1 antisense is associated with loss of tumorigenicity and potentiation of cytotoxicity to cisplatinum. J Clin Invest 1998; 101: 344–352.
Pagano M, Theodoras AM, Tam SW, Draetta GF. Cyclin D1-mediated inhibition of repair and replicative DNA synthesis in human fibroblasts Genes Dev 1994; 8: 1627–1639.
Han EK-H, Begemann M, Sgambato A, et al.. Increased expression of cyclin D1 in a murine mammary epithelial cell line induces p27kip1, inhibits growth, and enhances apoptosis. Cell Growth & Differ 1996; 7: 699–710.
Huang DCS, Strasser, S. BH3-Only proteins-essential initiators of apoptotic Cell death. Cell 2000; 103: 839–842.
Murphy KM, Streips UN, Lock RB. Bcl-2 Inhibits a Fas-induced Conformational Change in the Bax N Terminus and Bax Mitochondrial Translocation. J Biol Chem 2000; 275:17225–17228.
Leu CM, Chang C, Hu C. Epidermal growth factor suppresses staurosporine-induced apoptosis by inducing mcl-1 via the MAP kinase pathway. Oncogene 2000; 19: 1665–1673.
Henson ES, Gibson EM, Villanueva J, Bristow NA, Haney N, Gibson SB. Increased expression of Mcl-1 is responsible for the blockage of TRAIL-induced apoptosis mediated by EGF/ErbB1 signaling pathway. J Cell Biochem 2003; 89: 1177–1192.
Alt JR, Cleveland JL, Hannink M, Diehl JA. Phosphorylation-dependent regulation of cyclin D1 nuclear export and cyclin D1-dependent cellular transformation. Genes Dev 2000; 14: 3102–3114
Kwok TT, Mok CH, Menton-Brennan L. Up-regulation of a mutant form of p53 by doxorubicin in Human Squamous Carcinoma cells. Cancer Res 1994; 54:2834–2836.
Chen J, Martin BL, Brautigan DL. Regulation of protein serine-threonine phosphatase type-2A by tyrosine phosphorylation. Science 1992; 257: 1261–126.
Quijano VJ Jr, Sheffield LG. Prolactin decreases epidermal growth factor receptor kinase activity via a phosphorylation-dependent mechanism. J Biol Chem 1998; 273: 1200–1207
Barbier AJ, Poppleton HM, Yigzaw Y, et al. Transmodulation of epidermal growth factor receptor function by cyclic AMP-dependent protein kinase. J Biol Chem 1999; 274: 14067–14073.
Winston JT, Olashaw NE, Pledger WJ. Regulation of the transmodulated epidermal growth factor receptor by cholera toxin and the protein phosphatase inhibitor okadaic acid. J Cell Biochem 1991; 47: 79–89.
Strasberg Rieber M, Rieber M. Accesibility to DNA in Carcinoma chromatin is promoted by nanomolar okadaic acid: effect on A-T rich DNA binding proteins. Cancer Res 1992; 52: 6397–6399
Sheikh MS, Garcia M, Zhan Q, Liu Y, Fornace AJ Jr. Cell cycle-independent regulation of p21Waf1/Cip1 and Retinoblastoma protein during okadaic acid-induced apoptosis is coupled with induction of bax protein in human breast carcinoma cells. Cell Growth Differ 1996; 7: 1599–1607
Connell-Crowley L, Elledge SJ, Harper JW. G1 cyclin-dependent kinases are sufficient to initiate DNA synthesis in quiescent human fibroblasts. Curr Biol 1998; 8: 65–68.
Strasberg Rieber M, Rieber M. Sensitization to DNA damage by okadaic acid or bromodeoxyuridine involves unequal effects on melanoma cell adhesion and differentiation. DNA & Cell Biology 1997; 16: 121–125
Strasberg Rieber M, Zangemeister-Wittke U, Rieber M. p53 independent induction of apoptosis in human melanoma cells by a bcl-2/bcl XL bispecific antisense oligonucleotide. Clinical Cancer Research 2001; 7: 1446–1451
O'Brien JO, Wilson I, Orton T, Pognan F. Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cytotoxicity. Eur J Biochem 2000; 267: 5421–5426.
Simbulan-Rosenthal CM, Rosenthal DS, Iyer S, Boulares H, Smulson ME. Involvement of PARP and poly(ADP-ribosyl)ation in the early stages of apoptosis and DNA replication. Mol Cell Biochem 1999; 193: 137–148
Hu Y, Dragowska WH, Wallis A, Duronio V, Mayer L. Cytotoxicity induced by manipulation of signal transduction pathways is associated with down-regulation of Bcl-2 but not Mcl-1 in MCF-7 human breast cancer. Breast Cancer Res Treat 2001; 70: 11–20.
Benito A, Lerga A, Silva M, Leon J, Fernandez-Luna JL. Apoptosis of human myeloid Leukemia cells induced by an inhibitor of protein phosphatases (okadaic acid) is prevented by Bcl-2 and Bcl-X(L). Leukemia 1997; 11: 940–944
Riordan FA, Foroni L, Hoffbrand AV, Mehta AB, Wickremasinghe RG. Okadaic acid-induced apoptosis of HL60 leukemia cells is preceded by destabilization of bcl-2 mRNA and downregulation of bcl-2 protein. FEBS Lett 1998; 435: 195–198
Yasuda Y, Yoshinaga N, Murayama T, Nomura Y. Inhibition of hydrogen peroxide induced apoptosis but not arachidonic acid release in GH3 cells by EGF. Brain Res 1999; 850: 197–206
Diehl JA, Cheng M, Roussel MF, Sherr CJ. Glycogen synthase kinase-3 beta regulates cyclin D1 proteolysis and subcellular localization. Genes Dev 1998; 12: 3499–3511.
Tan A, Bitterman P, Sonenberg N, Peterson M, Polunovsky V. Inhibition of Myc-dependent apoptosis by eukaryotic translation initiation factor 4E requires cyclin D1. Oncogene 2000; 19: 1437–1447.
Yam CH, Siu WY, Lau A, Poon RC. degradation of cyclin A does not require its phosphorylation by cdc2 and cyclin-dependent kinase 2. J Biol Chem 2000; 275: 3158–3167.
Lin HM, Lee YJ, Li G, Pestell RG, Kim HR. Bcl-2 induces cyclin D1 promoter activity in human breast epithelial cells independent of cell anchorage. Cell Death Differ 2001; 8: 44–50.
Steeg P, Zhou Q. Cyclins and breast cancer. Breast Cancer Res Treat 1998; 52: 17.
Mueller A, Odze R, Jenkins TD, et al. A transgenic mouse model with cyclin D1 overexpression results in cell cycle, epidermal growth factor receptor,and p53 abnormalities. Cancer Res 1997; 57: 5542–5549.
Mineta H, Borg A, Dictor M, Wahlbert P, Wennerberg J. Correlation between p53 mutation and cyclin D1 amplification in head and neck squamous cell carcinoma. Oral Oncol 1997; 33: 42–46.
Rieber M, Strasberg Rieber M. Tumor suppression without differentiation of apoptosis by antisense cyclin D1 gene transfer in K1735 melanoma involves induction of p53, p21WAF1 and superoxide dismutase. Cell Death Diff 1999; 6: 1209–1215.
Milas L, Akimoto T, Hunter NR, et al. Relationship between cyclin D1 expression and poor radioresponse of murine carcinomas. Int J Radiat Oncol Biol Phys 2002; 52: 514–521.
Strasberg RM, Anzellotti A, Sanchez-Delgado R, Y Rieber M. Tumor apoptosis induced by ruthenium(II)-ketoconazole is enhanced in nonsusceptible carcinoma by monoclonal antibody to EGF receptor. Int J Cancer 2004; 112(3): 376–384.
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Rieber, M., Rieber, M.S. Cyclin D1 overexpression induces epidermal growth factor-independent resistance to apoptosis linked to BCL-2 in human A431 carcinoma. Apoptosis 11, 121–129 (2006). https://doi.org/10.1007/s10495-005-3084-4
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DOI: https://doi.org/10.1007/s10495-005-3084-4