Abstract
p53 tumour-suppressor gene is involved in cell growth control, arrest and apoptosis. Nevertheless cell cycle arrest and apoptosis induction can be observed in p53-defective cells after exposure to DNA-damaging agents such as 5-fluorouracil (5-FU) suggesting the importance of alternative pathways via p53-independent mechanisms. In order to establish relationship between p53 status, cell cycle arrest, Bcl-2/Bax regulation and 5-FU sensitivity, we examined p53 mRNA and protein expression and p53 protein functionality in wild-type (wt) and mutant (mt) p53 cell lines. p53 mRNA and p53 protein expression were determined before and after exposure to equitoxic 5-FU concentration in six human carcinoma cell lines differing in p53 status and displaying marked differences in 5-FU sensitivity, with IC50values ranging from 0.2–22.6 mM. 5-FU induced a rise in p53 mRNA expression in mt p53 cell lines and in human papilloma virus positive wt p53 cell line, whereas significant decrease in p53 mRNA expression was found in wt p53 cell line. Whatever p53 status, 5-FU altered p53 transcriptional and translational regulation leading to up-regulation of p53 protein. In relation with p53 functionality, but independently of p53 mutational status, after exposure to 5-FU equitoxic concentration, all cell lines were able to arrest in G1. No relationship was evidenced between G1 accumulation ability and 5-FU sensitivity. Moreover, after 5-FU exposure, Bax and Bcl-2 proteins regulation was under p53 protein control and a statistically significant relationship (r= 0.880,P= 0.0097) was observed between Bcl-2/Bax ratio and 5-FU sensitivity. In conclusion, whatever p53 status, Bcl-2 or Bax induction and Bcl-2/Bax protein ratio were correlated to 5-FU sensitivity. © 2000 Cancer Research Campaign
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Aguilar Santelises M, Rottenberg ME, Lewin N, Mellstedt H and Jondal M (1996) Bcl-2, Bax and p53 expression in B-CLL in relation to in vitro survival and clinical progression. Int J Cancer 69: 114–119
Ara S, Lee PS, Hansen MF and Saya H (1990) Codon 72 polymorphism of the TP53 gene. Nucleic Acids Res 18: 4961
Arrowsmith CH and Morin P (1996) New insights into p53 function from structural studies. Oncogene 12: 1379–1385
Barak Y, Juven T, Haffner R and Oren M (1993) mdm2 expression is induced by wild type p53 activity. EMBO J 12: 461–468
Barak Y, Gottlieb E, Juven Gershon T and Oren M (1994) Regulation of mdm2 expression by p53: alternative promoters produce transcripts with nonidentical translation potential. Genes Dev 8: 1739–1749
Barberi-Heyob M, Griffon G, Merlin JL and Weber B (1993) Sequence-dependent growth-inhibitory effects of the in vitro combination of fluorouracil, cisplatin and dipyridamole. Cancer Chemother Pharmacol 33: 163–170
Beck A, Etienne MC, Cheradame S, Fischel JL, Formento P, Renee N and Milano G (1994) A role for dihydropyrimidine dehydrogenase and thymidylate synthase in tumour sensitivity to fluorouracil. Eur J Cancer 30a: 1517–1522
Boshart M, Gissmann L, Ikenberg H, Kleinheinz A, Scheurlen W and zur Hausen H (1984) A new type of papillomavirus DNA, its presence in genital cancer biopsies and in cell lines derived from cervical cancer. EMBO J 3: 1151–1157
Cho Y, Gorina S, Jeffrey PD and Pavletich NP (1994) Crystal structure of a p53 tumor suppressor – DNA complex: understanding tumorigenic mutations [see comments]. Science 265: 346–355
Chou TC and Talalay P (1987) Application of the median-effect principle for the assessment of low dose risk of carcinogens and for the quantitation of synergism and antagonism of chemotherapeutic agents. In New Avenues in Developmental Cancer Chemotherapy, Harrap KR, Connors TA (eds) pp. 37–64, Academic Press: New York
Crook T, Tidy JA and Vousden KH (1991) Degradation of p53 can be targeted by HPV E6 sequences distinct from those required for p53 binding and trans-activation. Cell 67: 547–556
Deffie A, Wu H, Reinke V and Lozano G (1993) The tumor suppressor p53 regulates its own transcription. Mol Cell Biol 13: 3415–3423
Dou QP, An B and Will PL (1995) Induction of a retinoblastoma phosphatase activity by anticancer drugs accompanies p53-independent G1 arrest and apoptosis. Proc Natl Acad Sci USA 92: 9019–9023
Elbendary AA, Cirisano FD, Evans AC Jr, Davis PL, Iglehart JD, Marks JR and Berchuck A (1996) Relationship between p21 expression and mutation of the p53 tumor suppressor gene in normal and malignant ovarian epithelial cells. Clin Cancer Res 2: 1571–1575
el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW and Vogelstein B (1993) WAF1, a potential mediator of p53 tumor suppression. Cell 75: 817–825
el-Deiry WS, Harper JW, O'Connor PM, Velculescu VE, Canman CE, Jackman J, Pietenpol JA, Burrell M, Hill DE and Wang Y (1994) WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res 54: 1169–1174
Etienne MC, Pivot X, Formento JL, Bensadoun RJ, Formento P, Dassonville O, Francoual M, Poissonnet G, Fontana X, Schneider M, Demard F and Milano G (1999) A multifactorial approach including tumoural epidermal growth factor receptor, p53, thymidylate synthase and dihydropyrimidine dehydrogenase to predict treatment outcome in head and neck cancer patients receiving 5-fluorouracil. Br J Cancer 79: 1864–1869
Ewen ME and Miller SJ (1996) p53 and translational control. Biochim Biophys Acta 1242: 181–184
Fisher TC, Milner AE, Gregory CD, Jackman AL, Aherne GW, Hartley JA, Dive C and Hickman JA (1993) bcl-2 modulation of apoptosis induced by anticancer drugs: resistance to thymidylate stress is independent of classical resistance pathways. Cancer-Res 53: 3321–3326
Fu L, Minden MD and Benchimol S (1996) Translational regulation of human p53 gene expression. EMBO J 15: 4392–4401
Gorgoulis VG, Zacharatos PV, Manolis E, Ikonomopoulos JA, Damalas A, Lamprinopoulos C, Rassidakis GZ, Zoumpourlis V, Kotsinas A, Rassidakis AN, Halazonetis TD and Kittas C (1998) Effects of p53 mutants derived from lung carcinomas on the p53-responsive element (p53RE) of the MDM2 gene. Br J Cancer 77: 374–384
Guillouf C, Grana X, Selvakumaran M, De Luca A, Giordano A, Hoffman B and Liebermann DA (1995) Dissection of the genetic programs of p53-mediated G1 growth arrest and apoptosis: blocking p53-induced apoptosis unmasks G1 arrest. Blood 85: 2691–2698
Gussow D, Rein R, Ginjaar I, Hochstenbach F, Seemann G, Kottman A and Ploegh HL (1987) The human beta 2-microglobulin gene. Primary structure and definition of the transcriptional unit. J Immunol 139: 3132–3138
Harris CC (1996) Structure and function of the p53 tumor suppressor gene: clues for rational cancer therapeutic strategies. J Natl Cancer Inst 88: 1442–1455
Hudson JM, Frade R and Bar Eli M (1995) Wild-type p53 regulates its own transcription in a cell-type specific manner. DNA Cell Biol 14: 759–766
Huibregtse JM, Scheffner M and Howley PM (1993) Cloning and expression of the cDNA for E6-AP, a protein that mediates the interaction of the human papillomavirus E6 oncoprotein with p53. Mol Cell Biol 13: 775–784
Kastan MB, Onyekwere O, Sidransky D, Vogelstein B and Craig RW (1991) Participation of p53 protein in the cellular response to DNA damage. Cancer Res 51: 6304–6311
Kirihara Y, Yamamoto W, Toge T and Nishiyama M (1999) Dihydropyrimidine dehydrogenase, multidrug resistance-associated protein, and thymidylate synthase gene expression levels can predict 5-fluorouracil resistance in human gastrointestinal cancer cells. Int J Oncol 14: 551–556
Koshiji M, Adachi Y, Taketani S, Takeuchi K, Hioki K and Ikehara S (1997) Mechanisms underlying apoptosis induced by combination of 5-fluorouracil and interferon-gamma. Biochem Biophys Res Commun 240: 376–381
Kren BT, Trembley JH and Steer CJ (1996) Alterations in mRNA stability during rat liver regeneration. Am J Physiol 270: G763–777
Kubbutat MH, Jones SN and Vousden KH (1997) Regulation of p53 stability by Mdm2. Nature 387: 299–303
Kuerbitz SJ, Plunkett BS, Walsh WV and Kastan MB (1992) Wild-type p53 is a cell cycle checkpoint determinant following irradiation. Proc Natl Acad Sci USA 89: 7491–7495
Loignon M, Fetni R, Gordon AJ and Drobetsky EA (1997) A p53-independent pathway for induction of p21 waflcip1 and concomitant G1 arrest in UV-irradiated human skin fibroblasts. Cancer Res 57: 3390–3394
Lowe SW, Ruley HE, Jacks T and Housman DE (1993) p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell 74: 957–967
Macleod KF, Sherry N, Hannon G, Beach D, Tokino T, Kinzler K, Vogelstein B and Jacks T (1995) p53-dependent and independent expression of p21 during cell growth, differentiation, and DNA damage. Genes Dev 9: 935–944
Massimi P and Banks L (1997) Repression of p53 transcriptional activity by the HPV E7 proteins. Virology 227: 255–259
Matlashewski GJ, Tuck S, Pim D, Lamb P, Schneider J and Crawford LV (1987) Primary structure polymorphism at amino acid residue 72 of human p53. Mol Cell Biol 7: 961–963
Moll UM, Ostermeyer AG, Haladay R, Winkfield B, Frazier M and Zambetti G (1996) Cytoplasmic sequestration of wild-type p53 protein impairs the G1 checkpoint after DNA damage. Mol Cell Biol 16: 1126–1137
Momand J, Zambetti GP, Olson DC, George D and Levine AJ (1992) The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation. Cell 69: 1237–1245
Mosner J, Mummenbrauer T, Bauer C, Sczakiel G, Grosse F and Deppert W (1995) Negative feedback regulation of wild-type p53 biosynthesis. EMBO J 14: 4442–4449
Nabeya Y, Loganzo F Jr, Maslak P, Lai L, de Oliveira AR, Schwartz GK, Blundell ML, Altorki NK, Kelsen DP and Albino AP (1995) The mutational status of p53 protein in gastric and esophageal adenocarcinoma cell lines predicts sensitivity to chemotherapeutic agents. Int J Cancer 64: 37–46
Nita ME, Nagawa H, Tominaga O, Tsuno N, Fujii S, Sasaki S, Fu CG, Takenoue T, Tsuruo T and Muto T (1998a) 5-Fluorouracil induces apoptosis in human colon cancer cell lines with modulation of Bcl-2 family proteins. Br J Cancer 78: 986–992
Nita ME, Tominaga O, Nagawa H, Tsuruo T and Muto T (1998b) Dihydropyrimidine dehydrogenase but not thymidylate synthase expression is associated with resistance to 5-fluorouracil in colorectal cancer. Hepatogastroenterology 45: 2117–2122
O'Connor PM, Jackman J, Jondle D, Bhatia K, Magrath I and Kohn KW (1993) Role of the p53 tumor suppressor gene in cell cycle arrest and radiosensitivity of Burkitt's lymphoma cell lines. Cancer Res 53: 4776–4780
Oltvai ZN, Milliman CL and Korsmeyer SJ (1993) Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74: 609–619
Oren M (1994) Relationship of p53 to the control of apoptotic cell death. Semin Cancer Biol 5: 221–227
Ory K, Legros Y, Auguin C and Soussi T (1994) Analysis of the most representative tumour-derived p53 mutants reveals that changes in protein conformation are not correlated with loss of transactivation or inhibition of cell proliferation. EMBO J 13: 3496–3504
Palmer DG, Paraskeva C and Williams AC (1997) Modulation of p53 expression in cultured colonic adenoma cell lines by the naturally occurring lumenal factors butyrate and deoxycholate. Int J Cancer 73: 702–706
Peters GJ, van der Wilt CL, van Triest B, Codacci-Pisanelli G, Johnston PG, van Groeningen CJ and Pinedo HM (1995) Thymidylate synthase and drug resistance. Eur J Cancer 31A: 1299–1305
Pinedo HM and Peters GF (1988) Fluorouracil: biochemistry and pharmacology. J Clin Oncol 6: 1653–1664
Reed JC (1994) Bcl-2 and the regulation of programmed cell death. J Cell Biol 124: 1–6
Scheffner M, Werness BA, Huibregtse JM, Levine AJ and Howley PM (1990) The E6 oncoprotein encoded by human papillomavirus types 16 and 18 promotes the degradation of p53. Cell 63: 1129–1136
Selvakumaran M, Lin HK, Miyashita T, Wang HG, Krajewski S, Reed JC, Hoffman B and Liebermann D (1994) Immediate early up-regulation of bax expression by p53 but not TGF beta 1: a paradigm for distinct apoptotic pathways. Oncogene 9: 1791–1798
Simonian PL, Grillot DA and Nunez G (1997) Bcl-2 and Bcl-XL can differentially block chemotherapy-induced cell death. Blood 90: 1208–1216
Spears CP, Gustavsson BG, Berne M, Frösing R, Bernstein L and Hayes AA (1988) Mechanisms of innate resistance to thymidylate synthase inhibition after 5-fluorouracil. Cancer Res 48: 5894–5900
Stewart N, Hicks GG, Paraskevas F and Mowat M (1995) Evidence for a second cell cycle block at G2/M by p53. Oncogene 10: 109–115
Strobel T, Swanson L, Korsmeyer S and Cannistra SA (1996) BAX enhances paclitaxel-induced apoptosis through a p53-independent pathway. Proc Natl Acad Sci USA 93: 14094–14099
Vogelstein B and Kinzler KW (1992) p53 function and dysfunction. Cell 70: 523–526
Wouters BG, Denko NC, Giaccia AJ and Brown JM (1999) A p53 and apoptotic-independent role for p21 waf1 in tumour response to radiation therapy. Oncogene 18: 6540–6545
Zauberman A, Flusberg D, Haupt Y, Barak Y and Oren M (1995) A functional p53-responsive intronic promoter is contained within the human mdm2 gene. Nucleic Acids Res 23: 2584–2592
Zhan Q, Fan S, Bae I, Guillouf C, Liebermann DA, O'Connor PM and Fornace AJ Jr (1994) Induction of bax by genotoxic stress in human cells correlates with normal p53 status and apoptosis (published erratum appears in Oncogene 1995; 10 (6): 1259). Oncogene 9: 3743–3751
Zhang ZG, Harstrick A and Rustum YM (1992) Mechanisms of resistance to fluoropyrimidines. Semin Oncol 19: 4–9
Author information
Authors and Affiliations
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Mirjolet, JF., Barberi-Heyob, M., Didelot, C. et al. Bcl-2/Bax protein ratio predicts 5-fluorouracil sensitivity independently of p53 status. Br J Cancer 83, 1380–1386 (2000). https://doi.org/10.1054/bjoc.2000.1455
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1054/bjoc.2000.1455
Keywords
This article is cited by
-
Re-sensitization of 5-FU resistance by SPARC through negative regulation of glucose metabolism in hepatocellular carcinoma
Tumor Biology (2015)
-
A Double-modulation Strategy in Cancer Treatment With a Chemotherapeutic Agent and siRNA
Molecular Therapy (2011)
-
Quantitative detection of ING1 mRNA under different gene regulation based on molecular beacon
Chinese Science Bulletin (2006)
-
Genetic factors influencing Pyrimidine-antagonist chemotherapy
The Pharmacogenomics Journal (2005)
-
Targeting BCL-2 overexpression in various human malignancies through NF-κB inhibition by the proteasome inhibitor bortezomib
Cancer Chemotherapy and Pharmacology (2005)