title: Staurosporine is chemoprotective by inducing G1 arrest in a Chk1- and pRb-dependent manner.
abstract: Chemotherapeutic agents have been the mainstay of cancer therapy for years. However, their effectiveness has been limited by toxicities they impart on normal cells. Staurosporine (ST) has been shown to arrest normal, but not breast cancer, cells in G1. Therefore, ST may become a chemoprotective agent, arresting normal cells while allowing tumor cells to enter cell cycle phases where they are sensitive to chemotherapeutic agents. Understanding the mechanism of ST-mediated G1 arrest may allow for a beneficial chemoprotective treatment strategy for patients. We utilized 76NE6 (pRb+/p53-), 76NF2V (pRb+/p53+) and 76NE7 (pRb-/P53+) non-tumorigenic human mammary epithelial cell lines to understand the role of the Rb and p53 pathways in ST-directed G1 arrest. CDK4 was downregulated by ST in Rb+ cells, but its presence could not reverse the arrest, neither did its stable downregulation alter ST-mediated cellular response. ST-mediated G1 arrest required pRb, which in turn initiated a cascade of events leading to inhibition of CDK4. Further assessment of this pathway revealed that Chk1 expression and activity were required for the Rb-dependent arrest. For example, pRb+ cells with small interfering RNA to Chk1 had approximately 60% less cells in G1 phase compared with controls and pRb- cells do not arrest upon ST. Furthermore, Chk1 expression facilitates the release of the Rb+ cells from G1 arrest. Collectively, our data suggest that pRb cooperates with Chk1 to mediate a G1 arrest only in pRb+ cells. The elucidation of this pathway can help identify novel agents to protect cancer patients against the debilitating effects of chemotherapy.
authors.affiliation: Department of Systems Biology and.
authors.forename: Mollianne McGahren
authors.initials: MM
authors.lastname: Murray
authors.forename: Tuyen
authors.initials: T
authors.lastname: Bui
authors.forename: Michelle
authors.initials: M
authors.lastname: Smith
authors.forename: Rozita
authors.initials: R
authors.lastname: Bagheri-Yarmand
authors.forename: Hannah
authors.initials: H
authors.lastname: Wingate
authors.forename: Kelly K
authors.initials: KK
authors.lastname: Hunt
authors.forename: Khandan
authors.initials: K
authors.lastname: Keyomarsi
mesh_terms: D059447:Cell Cycle Checkpoints / Q000187:drug effects / Q000235:genetics; D002461:Cell Line, Transformed; D000071877:Checkpoint Kinase 1; D051358:Cyclin-Dependent Kinase 4 / Q000235:genetics / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D059585:G1 Phase Cell Cycle Checkpoints / Q000187:drug effects* / Q000502:physiology*; D015870:Gene Expression; D006801:Humans; D020011:Protective Agents / Q000494:pharmacology*; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D016160:Retinoblastoma Protein / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D019311:Staurosporine / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; D020011:Protective Agents; D016160:Retinoblastoma Protein; D011494:Protein Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1; D051358:Cyclin-Dependent Kinase 4; D019311:Staurosporine