title: The DNA-binding epidermal growth factor-receptor inhibitor PD153035 and other DNA-intercalating cytotoxic drugs reactivate the expression of the retinoic acid receptor-beta tumor-suppressor gene in breast cancer cells.
abstract: We have previously shown that the epidermal growth factor-receptor (EGFR) tyrosine kinase inhibitor PD153035 induces retinoic acid receptor-beta (RAR-beta) expression in malignant cells by mechanisms that are independent of its blocking activity on EGFR (ErbB1) or on any other ErbB receptor (ErbB2, ErbB3, ErbB4). RAR-beta2, one of three human RAR-beta isoforms (RAR-beta1, RAR-beta2, RAR-beta4), is silenced in many tumors and acts as a tumor suppressor. Forced expression of RAR-beta2 reverts the malignant phenotype of RAR-beta2-negative breast cancer cells and reconstitutes retinoid sensitivity in these cells. Here, we demonstrate that the EGFR inhibitor PD153035 specifically induces RAR-beta2, but not the other two isoforms (RAR-beta1, RAR-beta4) in MDA-MB-468 and MDA-MB-453 human breast cancer cells. Induction was seen at the mRNA (reverse transcription-polymerase chain reaction) and protein level (Western analysis). PD153035-mediated induction of RAR-beta2 was associated with synergistic growth inhibition in cells co-treated with PD153035 and all-trans retinoic acid (tRA). Most importantly, PD153035 restored retinoic acid sensitivity in retinoic acid-resistant cells. Our previous work also revealed that PD153035 directly intercalates into the DNA suggesting that changes in the chromatin structure contribute to the RAR-beta2-inducing effect of PD153035. This prompted us to examine the effect of DNA intercalating chemotherapeutic drugs such as doxorubicin, amsacrine, and mitoxantrone on the expression of RAR-beta. Vincristine was used for comparative reasons, because this drug does not target DNA. All four compounds caused dose-dependent growth inhibition in MDA-MB-468 and MDA-MB-453 cells. Interestingly, compounds that directly interact with the DNA (doxorubicin, amsacrine, mitoxantrone) caused a time-dependent up-regulation of the RAR-beta expression in all cell lines examined, whereas the negative control drug vincristine, which causes disruption of microtubule structures, did not stimulate RAR-beta expression. These data further support the notion that induction of the RAR-beta tumor-suppressor gene in cancer cells by PD153035 is mediated at least in part by its DNA intercalating activity.
authors.affiliation: Signaling Networks Program, Department of Medicine I, Division of Oncology, Medical University of Vienna, Vienna, Austria. thomas.grunt@meduniwien.ac.at
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Grunt
authors.forename: Katharina
authors.initials: K
authors.lastname: Tomek
authors.forename: Renate
authors.initials: R
authors.lastname: Wagner
authors.forename: Klaudia
authors.initials: K
authors.lastname: Puckmair
authors.forename: Christoph C
authors.initials: CC
authors.lastname: Zielinski
mesh_terms: D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000201:enzymology / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D066246:ErbB Receptors / Q000037:antagonists & inhibitors*; D016147:Genes, Tumor Suppressor / Q000187:drug effects*; D006801:Humans; D007364:Intercalating Agents / Q000378:metabolism / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D011799:Quinazolines / Q000378:metabolism / Q000494:pharmacology*; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D000970:Antineoplastic Agents; D007364:Intercalating Agents; D047428:Protein Kinase Inhibitors; D011799:Quinazolines; D018168:Receptors, Retinoic Acid; C082925:retinoic acid receptor beta; D004247:DNA; D066246:ErbB Receptors; C088860:4-((3-bromophenyl)amino)-6,7-dimethoxyquinazoline