title: Multifactorial analysis of conditional reprogramming of human keratinocytes.
abstract: Co-culture of human primary epithelial cells with irradiated 3T3 fibroblast feeder cells (J2 cells) and the Rho kinase inhibitor Y-27632 (Y) allows for the unrestricted growth of cells of epithelial origin by the process termed conditional reprogramming. To better understand the nature of the signaling processes associated with conditionally reprogrammed cells, the effect of the two critical components of the co-culture conditions, J2 cells and Y, on the growth of human foreskin keratinocytes (HFKs) was evaluated by gene expression profiling, reverse-phase protein arrays and siRNA screening. J2 cells and Y acted cooperatively to down-regulate differentiation, and upregulate proliferation and cell adhesion, including increased pT308Akt and pERK, and reduced TGF-β pathway signaling. These findings establish a mechanistic basis for the unlimited growth potential of human epithelial cells that will be invaluable to assess the effect of genetic changes in pathologic tissues and their response to therapeutic agents.
authors.affiliation: Department of Pathology, Georgetown University, Washington, DC, 20007, United States of America.
authors.forename: Segni B
authors.initials: SB
authors.lastname: Ligaba
authors.affiliation: Department of Pathology, Georgetown University, Washington, DC, 20007, United States of America.
authors.forename: Anikita
authors.initials: A
authors.lastname: Khurana
authors.affiliation: Department of Oncology, Georgetown University, Washington, DC, 20007, United States of America.
authors.forename: Garrett
authors.initials: G
authors.lastname: Graham
authors.affiliation: Department of Pathology, Georgetown University, Washington, DC, 20007, United States of America.
authors.forename: Ewa
authors.initials: E
authors.lastname: Krawczyk
authors.affiliation: Department of Oncology, Georgetown University, Washington, DC, 20007, United States of America; Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, 20007, United States of America.
authors.forename: Sandra
authors.initials: S
authors.lastname: Jablonski
authors.affiliation: Department of Molecular and Microbiology, George Mason University, Manassas, Virginia, 20110, United States of America.
authors.forename: Emanuel F
authors.initials: EF
authors.lastname: Petricoin
authors.affiliation: Department of Oncology, Georgetown University, Washington, DC, 20007, United States of America; Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, 20007, United States of America.
authors.forename: Robert I
authors.initials: RI
authors.lastname: Glazer
authors.affiliation: Department of Pathology, Georgetown University, Washington, DC, 20007, United States of America; Department of Oncology, Georgetown University, Washington, DC, 20007, United States of America; Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC, 20007, United States of America.
authors.forename: Geeta
authors.initials: G
authors.lastname: Upadhyay
mesh_terms: D000577:Amides / Q000494:pharmacology; D000818:Animals; D002448:Cell Adhesion; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D016000:Cluster Analysis; D018920:Coculture Techniques; D061252:Feeder Cells; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D015603:Keratinocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011725:Pyridines / Q000494:pharmacology; D034741:RNA, Small Interfering
chemical_list: D000577:Amides; D011725:Pyridines; D034741:RNA, Small Interfering; C108830:Y 27632