Wild-type p53 can induce p21 and apoptosis in neuroblastoma cells but the DNA damage-induced G1 checkpoint function is attenuated.

p53 is a tumor suppressor protein important in the regulation of apoptosis. Because p53 functions as a transcription factor, cellular responses depend upon activity of p53 localized in the nucleus. Cytoplasmic sequestration of p53 has been proposed as a mechanism by which the function of this protein can be suppressed, particularly in tumor types such as neuroblastoma in which the frequency of mutations of p53 is low. Data presented here demonstrate that nuclear p53 protein is expressed in a panel of neuroblastoma cell lines, and after exposure to DNA damage, transcriptionally active p53 expression can be induced. After exposure to both equitoxic IC80 and 10-Gy doses of ionizing radiation, both p53 and p21 were induced, but G1 cell cycle arrest was attenuated. To investigate whether the DNA damage signaling pathway was incapable of inducing sufficient p53 in these cells, we expressed additional wild-type p53 after adenoviral vector transduction. This exogenous p53 expression also resulted in p21 induction but was unable to enhance the G1 arrest, suggesting that the pathway downstream from p53 is nonfunctional. Although p53-mediated G1 arrest is attenuated in neuroblastoma cells, the ability of p53 to induce apoptosis appears functional, consistent with its chemosensitive phenotype. This work demonstrates that p53 is expressed in the nucleus of neuroblastoma cells and can mediate induction of p21. However, this cell type appears to have an attenuated ability to mediate a DNA damage-induced G1 cell cycle arrest.

[1]  D. Conrad,et al.  Dysregulation of the cyclin-dependent kinase inhibitor p21WAF1/CIP1/MDA6 increases the susceptibility of human leukemia cells (U937) to 1-beta-D-arabinofuranosylcytosine-mediated mitochondrial dysfunction and apoptosis. , 1999, Cancer research.

[2]  P. Fisher,et al.  Dysregulation of the Cyclin-dependent Kinase Inhibitor p21 the Susceptibility of Human Leukemia Cells (U937) to 1- b - D -Arabinofuranosylcytosine-mediated Mitochondrial Dysfunction and Apoptosis 1 , 1999 .

[3]  M. Strauss,et al.  p21WAF1/CIP1 mutants deficient in inhibiting cyclin-dependent kinases (CDKs) can promote assembly of active cyclin D/CDK4(6) complexes in human tumor cells. , 1998, Cancer research.

[4]  R. Muschel,et al.  Radiation and the G2 phase of the cell cycle. , 1998, Radiation research.

[5]  M. Danks,et al.  Differences in epitope accessibility of p53 monoclonal antibodies suggest at least three conformations or states of protein binding of p53 protein in human tumor cell lines , 1998, Cell Death and Differentiation.

[6]  J. Barrett,et al.  Differential subcellular p53 localization and function in N- and S-type neuroblastoma cell lines. , 1998, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[7]  J. Easton,et al.  Disruption of the cyclin D/cyclin-dependent kinase/INK4/retinoblastoma protein regulatory pathway in human neuroblastoma. , 1998, Cancer research.

[8]  M. Kasten,et al.  pRb and the Cdks in apoptosis and the cell cycle , 1998, Cell Death and Differentiation.

[9]  J. Houghton,et al.  Bax is an important determinant of chemosensitivity in pediatric tumor cell lines independent of Bcl-2 expression and p53 status. , 1998, Oncology research.

[10]  P. Houghton,et al.  Efficacy of systemic administration of irinotecan against neuroblastoma xenografts. , 1997, Clinical cancer research : an official journal of the American Association for Cancer Research.

[11]  G. Hannon,et al.  Functional Analysis of a p21WAF1,CIP1,SDI1 Mutant (Arg94 → Trp) Identified in a Human Breast Carcinoma , 1996, The Journal of Biological Chemistry.

[12]  Bert Vogelstein,et al.  Uncoupling of S phase and mitosis induced by anticancer agents in cells lacking p21 , 1996, Nature.

[13]  H. Shepard,et al.  Gene transfer into human bone marrow hematopoietic cells mediated by adenovirus vectors. , 1996, Blood.

[14]  R. Beijersbergen,et al.  Cell cycle regulation by the retinoblastoma family of growth inhibitory proteins. , 1996, Biochimica et biophysica acta.

[15]  M. Kastan,et al.  The p53 signal transduction pathway is intact in human neuroblastoma despite cytoplasmic localization. , 1996, The American journal of pathology.

[16]  M. Oren,et al.  Abrogation of wild-type p53 mediated growth-inhibition by nuclear exclusion. , 1996, Oncogene.

[17]  M. Danks,et al.  Subcellular redistribution of DNA topoisomerase I in anaplastic astrocytoma cells treated with topotecan. , 1996, Cancer research.

[18]  U. Moll,et al.  Cytoplasmic sequestration of wild-type p53 protein impairs the G1 checkpoint after DNA damage , 1996, Molecular and cellular biology.

[19]  D. Grignon,et al.  Somatic mutations of the WAF1/CIP1 gene in primary prostate cancer. , 1995, Oncogene.

[20]  G. Stark,et al.  p53 controls both the G2/M and the G1 cell cycle checkpoints and mediates reversible growth arrest in human fibroblasts. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Stephen J. Elledge,et al.  Mice Lacking p21 CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control , 1995, Cell.

[22]  C. Guillouf,et al.  p53 involvement in control of G2 exit of the cell cycle: role in DNA damage-induced apoptosis. , 1995, Oncogene.

[23]  K. Matthay.  Neuroblastoma: A clinical challenge and biologic puzzle , 1995, CA: a cancer journal for clinicians.

[24]  P. O'Connor,et al.  A mutant p21 cyclin-dependent kinase inhibitor isolated from a Burkitt's lymphoma. , 1995, Cancer research.

[25]  G. Hicks,et al.  Evidence for a second cell cycle block at G2/M by p53. , 1995, Oncogene.

[26]  K. Kohn,et al.  p53 gene mutations are associated with decreased sensitivity of human lymphoma cells to DNA damaging agents. , 1994, Cancer research.

[27]  K W Kohn,et al.  Cell cycle control and cancer chemotherapy , 1994, Journal of cellular biochemistry.

[28]  Stephen J. Elledge,et al.  p53-dependent inhibition of cyclin-dependent kinase activities in human fibroblasts during radiation-induced G1 arrest , 1994, Cell.

[29]  Yi-Song Wang,et al.  WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. , 1994, Cancer research.

[30]  R. Weinberg,et al.  Tumor spectrum analysis in p53-mutant mice , 1994, Current Biology.

[31]  J. Trent,et al.  WAF1, a potential mediator of p53 tumor suppression , 1993, Cell.

[32]  J. Pelletier,et al.  Absence of p53 gene mutations in primary neuroblastomas. , 1993, Cancer research.

[33]  K. Kohn,et al.  Role of the p53 tumor suppressor gene in cell cycle arrest and radiosensitivity of Burkitt's lymphoma cell lines. , 1993, Cancer research.

[34]  D. Housman,et al.  p53-dependent apoptosis modulates the cytotoxicity of anticancer agents , 1993, Cell.

[35]  A. Levine,et al.  A comparison of the biological activities of wild‐type and mutant p53 , 1993, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[36]  J. M. Lee,et al.  p53 mutations increase resistance to ionizing radiation. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[37]  Bert Vogelstein,et al.  Oncoprotein MDM2 conceals the activation domain of tumour suppressor p53 , 1993, Nature.

[38]  C. Purdie,et al.  Thymocyte apoptosis induced by p53-dependent and independent pathways , 1993, Nature.

[39]  P. Meltzer,et al.  Amplification of a gene encoding a p53-associated protein in human sarcomas , 1992, Nature.

[40]  D. Lane,et al.  p53, guardian of the genome , 1992, Nature.

[41]  A. Levine,et al.  The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation , 1992, Cell.

[42]  A. Davidoff,et al.  Expression of p53 in human neuroblastoma- and neuroepithelioma-derived cell lines. , 1992, Oncogene.

[43]  B. Vogelstein,et al.  Participation of p53 protein in the cellular response to DNA damage. , 1991, Cancer research.

[44]  A. Levine,et al.  The p53 tumour suppressor gene , 1991, Nature.

[45]  W. Crist,et al.  Common solid tumors of childhood. , 1991, The New England journal of medicine.

[46]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.