PMID- 10590214
OWN - NLM
STAT- MEDLINE
DCOM- 20000211
LR  - 20190513
IS  - 0143-3334 (Print)
IS  - 0143-3334 (Linking)
VI  - 20
IP  - 12
DP  - 1999 Dec
TI  - Effects of the rodent peroxisome proliferator and hepatocarcinogen, 
      perfluorooctanoic acid, on apoptosis in human hepatoma HepG2 cells.
PG  - 2237-46
AB  - The effects of perfluorooctanoic acid (PFOA), a potent hepatocarcinogen and 
      peroxisome proliferator in rodents, on human cells have not yet been examined. In 
      the present study we demonstrate that treatment of human hepatoblastoma HepG2 
      cells with PFOA induces apoptosis, as well as perturbs the cell cycle. This 
      apoptosis was characterized by electron microscopy, which revealed typical 
      nucleosomal fragmentation (also observed as a 'DNA ladder' upon electrophoresis 
      on agarose) and was quantitated using propidium iodide staining of cellular DNA 
      and the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling 
      (TUNEL) assay. This process was dose- and time-dependent: apoptosis became 
      manifest with 200 microM and maximal (45% of the cells) upon exposure to 450 
      microM PFOA for 24 h. Electrophoresis of the DNA from HepG2 cells exposed to 500 
      microM PFOA for 24 h or to 400 microM PFOA for 48 h revealed a smear typical of 
      non-specific degradation. These findings indicate that in the presence of high 
      concentrations of PFOA for long times, HepG2 cells undergo primary and secondary 
      necrosis. Quantitation of trypan blue exclusion supported this conclusion. Flow 
      cytometric analysis revealed that the cell cycle of HepG2 cells was perturbed by 
      exposure to 50-150 microM PFOA. A 50 microM concentration resulted in a 
      significant increase in the proportion of G(2)/M cells and, simultaneously, a 
      decrease in the number of cells in the S phase, whereas treatment with 100 or 150 
      microM PFOA increased the proportion of cells in the G(0)/G(1) phase and 
      decreased the number of cells in the G(2)/M and S phases. Simultaneous flow 
      cytometric analysis of apoptosis-associated DNA strand breaks using the TUNEL 
      procedure and of propidium iodide staining of cellular DNA revealed DNA breaks in 
      HepG2 cells exposed to 150 microM PFOA, prior to nuclear fragmentation.
FAU - Shabalina, I G
AU  - Shabalina IG
AD  - Unit of Biochemical Toxicology, Department of Biochemistry, Wallenberg 
      Laboratory, Stockholm University, S-106 91 Stockholm, Sweden.
FAU - Panaretakis, T
AU  - Panaretakis T
FAU - Bergstrand, A
AU  - Bergstrand A
FAU - DePierre, J W
AU  - DePierre JW
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Carcinogenesis
JT  - Carcinogenesis
JID - 8008055
RN  - 0 (Caprylates)
RN  - 0 (Carcinogens)
RN  - 0 (Fluorocarbons)
RN  - 0 (Peroxisome Proliferators)
RN  - 947VD76D3L (perfluorooctanoic acid)
SB  - IM
MH  - Apoptosis/*drug effects
MH  - Caprylates/*pharmacology
MH  - Carcinogens/*pharmacology
MH  - Carcinoma, Hepatocellular/*pathology/ultrastructure
MH  - Cell Survival/drug effects
MH  - Dose-Response Relationship, Drug
MH  - Electrophoresis, Agar Gel
MH  - Fluorocarbons/*pharmacology
MH  - Humans
MH  - In Situ Nick-End Labeling
MH  - Liver Neoplasms/*pathology/ultrastructure
MH  - Microscopy, Electron
MH  - Peroxisome Proliferators/*pharmacology
MH  - Tumor Cells, Cultured
EDAT- 1999/12/11 09:00
MHDA- 2000/02/19 09:00
CRDT- 1999/12/11 09:00
PHST- 1999/12/11 09:00 [pubmed]
PHST- 2000/02/19 09:00 [medline]
PHST- 1999/12/11 09:00 [entrez]
AID - 10.1093/carcin/20.12.2237 [doi]
PST - ppublish
SO  - Carcinogenesis. 1999 Dec;20(12):2237-46. doi: 10.1093/carcin/20.12.2237.