title: Drug-resistant breast cancer cells frequently retain expression of a functional wild-type p53 protein.
abstract: Abnormalities in the p53 tumor suppressor gene have been shown to affect cellular processes related to cell cycle control and gene amplification. In this study we compare the status and function of wild-type p53 in MCF-7 breast cancer cells with sublines selected for resistance to chemotherapeutic agents having different mechanisms of action. Sublines that were resistant to melphalan, pyrazafurin, mitoxantrone, etoposide and PALA all retained expression of wild-type p53. Methotrexate-resistant MCF-7 cells were unusual heterozygotes that expressed a wild-type and dominant, in-frame p53 deletion mutant and the doxorubicin-resistant cells expressed only mutant p53. Analysis of the G1 checkpoint after treatment with ionizing radiation revealed that the pyrazafurin-, melphalan- and mitoxantrone-resistant cells arrested strongly in G1. The etoposide- and PALA-resistant cells had an intermediate G1 arrest phenotype and the methotrexate- and doxorubicin-resistant cells had a minimal G1 arrest phenotype. mRNA and protein analyses of downstream effector genes, including P21CIP1/Waf1, mdm2, Gadd 45 and the retinoblastoma protein, did not entirely differentiate sublines having a strong versus intermediate G1 arrest phenotype. Neither the p53 status nor the strength of the G1 arrest could be correlated with cell survival after ionizing radiation. When drug-sensitive MCF-7 cells were treated with the same chemotherapeutic agents, p53 and p21CIP1/Waf1 levels increased between 2- and 14-fold. Together these data suggest that other cellular factors likely play a role in overcoming the inhibitory effects of ionizing radiation on p53 in drug-resistant breast cancer cells.
authors.affiliation: Division of Cancer Treatment, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: J M
authors.initials: JM
authors.lastname: Gudas
authors.forename: H
authors.initials: H
authors.lastname: Nguyen
authors.forename: T
authors.initials: T
authors.lastname: Li
authors.forename: L
authors.initials: L
authors.lastname: Sadzewicz
authors.forename: R
authors.initials: R
authors.lastname: Robey
authors.forename: K
authors.initials: K
authors.lastname: Wosikowski
authors.forename: K H
authors.initials: KH
authors.lastname: Cowan
mesh_terms: D000577:Amides; D000970:Antineoplastic Agents / Q000633:toxicity*; D001224:Aspartic Acid / Q000031:analogs & derivatives / Q000633:toxicity; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics*; D002453:Cell Cycle; D002999:Clone Cells; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000032:analysis / Q000096:biosynthesis; D019008:Drug Resistance, Neoplasm; D004791:Enzyme Inhibitors / Q000032:analysis; D005047:Etoposide / Q000633:toxicity; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression; D016158:Genes, p53; D006801:Humans; D007150:Immunohistochemistry; D008558:Melphalan / Q000633:toxicity; D008942:Mitoxantrone / Q000633:toxicity; D009363:Neoplasm Proteins / Q000032:analysis / Q000096:biosynthesis; D009687:Nuclear Proteins; D010746:Phosphonoacetic Acid / Q000031:analogs & derivatives / Q000633:toxicity; D016133:Polymerase Chain Reaction; D011518:Proto-Oncogene Proteins / Q000032:analysis / Q000096:biosynthesis; D051736:Proto-Oncogene Proteins c-mdm2; D011720:Pyrazoles; D012263:Ribonucleosides / Q000633:toxicity; D012266:Ribose; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000096:biosynthesis*
chemical_list: D000577:Amides; D000970:Antineoplastic Agents; C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D004791:Enzyme Inhibitors; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D011720:Pyrazoles; D012263:Ribonucleosides; D016159:Tumor Suppressor Protein p53; D001224:Aspartic Acid; C002997:pyrazofurin; D012266:Ribose; D005047:Etoposide; C013195:sparfosic acid; D008942:Mitoxantrone; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; D010746:Phosphonoacetic Acid; D008558:Melphalan