title: Aberrant cell cycle checkpoint function in transformed hepatocytes and WB-F344 hepatic epithelial stem-like cells.
abstract: Cell cycle checkpoints are barriers to carcinogenesis as they function to maintain genomic integrity. Attenuation or ablation of checkpoint function may enhance tumor formation by permitting outgrowth of unstable cells with damaged DNA. To examine the function of cell cycle checkpoints in rat hepatocarcinogenesis, we analyzed the responses of the G (1), G (2) and mitotic spindle assembly checkpoints in normal rat hepatocytes, hepatic epithelial stem-like cells (WB-F344) and transformed derivatives of both. Normal rat hepatocytes (NRH) displayed a 73% reduction in the fraction of nuclei in early S-phase 6-8 h following 8 Gy of ionizing radiation (IR) as a quantitative measure of G (1) checkpoint function. Chemically and virally transformed hepatocyte lines displayed significant attenuation of G (1) checkpoint function, ranging from partial to complete ablation. WB-F344 rat hepatic epithelial cell lines at low, mid and high passage levels expressed G (1) checkpoint function comparable with NRH. Only one of four malignantly transformed WB-F344 cell lines displayed significant attenuation of G (1) checkpoint function. Attenuation of G (1) checkpoint function in transformed hepatocytes and WB-F344 cells was associated with alterations in p53, ablated/attenuated induction of p21 (Waf1) by IR, as well as aberrant function of the spindle assembly checkpoint. NRH displayed 93% inhibition of mitosis 2 h after 1 Gy IR as a quantitative measure of G (2) checkpoint function. All transformed hepatocyte and WB-F344 cell lines displayed significant attenuation of the G (2) checkpoint. Moreover, the parental WB-F344 line displayed significant age-related attenuation of G (2) checkpoint function. Abnormalities in the function of cell cycle checkpoints were detected in transformed hepatocytes and WB-F344 cells at stages of hepatocarcinogenesis preceding tumorigenicity, sustaining a hypothesis that aberrant checkpoint function contributes to carcinogenesis.
authors.affiliation: Departments of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7295, USA. bill_kaufmann@med.unc.edu
authors.forename: W K
authors.initials: WK
authors.lastname: Kaufmann
authors.forename: C I
authors.initials: CI
authors.lastname: Behe
authors.forename: V M
authors.initials: VM
authors.lastname: Golubovskaya
authors.forename: L L
authors.initials: LL
authors.lastname: Byrd
authors.forename: C D
authors.initials: CD
authors.lastname: Albright
authors.forename: K M
authors.initials: KM
authors.lastname: Borchet
authors.forename: S C
authors.initials: SC
authors.lastname: Presnell
authors.forename: W B
authors.initials: WB
authors.lastname: Coleman
authors.forename: J W
authors.initials: JW
authors.lastname: Grisham
authors.forename: G J
authors.initials: GJ
authors.lastname: Smith
mesh_terms: D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle; D002461:Cell Line, Transformed; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000096:biosynthesis; D017931:DNA Primers; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008297:Male; D018807:Polymorphism, Single-Stranded Conformational; D051381:Rats; D011916:Rats, Inbred F344; D008941:Spindle Apparatus; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
chemical_list: C493958:Cdkn1a protein, rat; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D017931:DNA Primers; D016159:Tumor Suppressor Protein p53