title: Analyzing Ras-associated cell proliferation signaling.
abstract: Ras-dependent signaling is an important regulator of cell cycle progression, proliferation, senescence, and apoptosis. Several of the downstream effectors of Ras play dual roles in each of these processes. Under one set of conditions, they promote cell cycle progression and proliferation; yet, in a different paradigm, they drive cell cycle arrest and apoptosis. Furthermore, there is cross talk between certain downstream effectors of Ras including the PI3K-AKT and Raf-MEK-ERK pathways. Here we describe a series of experiments used to dissect the effect of different Ras-dependent signaling pathways on cell cycle progression, proliferation, senescence, and apoptosis. Furthermore, we highlight the importance of consistent growth conditions of cells in culture when studying Ras-dependent signaling as we show that the activation of downstream effectors of Ras changes with the confluency at which the cells are grown.
authors.affiliation: Department of Pharmacology and Physiology, Drexel University College of Medicine, 245 N. 15th Street, MS 488, Philadelphia, PA, 19102, USA.
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Stout
authors.forename: Edgar
authors.initials: E
authors.lastname: Asiimwe
authors.forename: James R
authors.initials: JR
authors.lastname: Birkenstamm
authors.forename: Su Yeon
authors.initials: SY
authors.lastname: Kim
authors.forename: Paul M
authors.initials: PM
authors.lastname: Campbell
mesh_terms: D000818:Animals; D023102:Anoikis; D001973:Bromodeoxyuridine / Q000032:analysis; D002453:Cell Cycle; D049109:Cell Proliferation; D016922:Cellular Senescence; D004586:Electrophoresis / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction; D013194:Staining and Labeling / Q000379:methods; D001616:beta-Galactosidase / Q000032:analysis* / Q000378:metabolism; D048490:raf Kinases / Q000378:metabolism; D018631:ras Proteins / Q000378:metabolism*
chemical_list: D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D048490:raf Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D001616:beta-Galactosidase; D018631:ras Proteins; D001973:Bromodeoxyuridine