2,4,4′-Trichlorobiphenyl increases STAT5 transcriptional activity

Greg G. Oakley, Amy L. Roe, Robert A. Blouin, Timothy P. Twaroski, Tanmoy C. Ganguly, Mary Vore, Hans Joachim Lehmler, Larry W. Robertson

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The promoting effects of polychlorinated biphenyls (PCBs) have been studied extensively in a variety of two-stage carcinogenesis models. However, the molecular mechanisms responsible for the promotion effects of PCBs have not been elucidated. We measured the effect of PCBs on DNA-binding proteins involved in cell proliferation and transformation. Male Sprague-Dawley rats were injected intraperitoneally with mono-, di-, tri-, tetra-, or hexachlorobiphenyls (300 μmol/kg/d) each day for 4 d and killed 4 h after the last injection. To detect alterations in nuclear proteins that could explain the tumor-promoter activity of PCBs, liver nuclear extracts were analyzed by electrophoretic mobility shift assays. Electrophoretic mobility shift assay analysis of signal transducers and activators of transcription (STAT)-binding activity to a consensus 7-interferon-activated sequence (GAS) element was compared in liver nuclear extracts from treated rats. STAT-binding activity was eightfold to tenfold higher in nuclear extracts from animals treated with 2,4,4′-trichloro- (PCB 28) and 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB 153). Analysis of the protein complex binding to the GAS element, with antibodies specific for STAT3, STAT5, and STAT6, indicated that the protein complex was made up of STAT5 and STAT6 proteins. HepG2 cells transiently transfected with a luciferase reporter gene construct containing many STAT5 binding sites were treated with PCB 28 and PCB 153. PCB 28 stimulated a greater than 25-fold increase in luciferase activity at the highest concentration tested, 1.0 μg/mL. However, enhanced luciferase activity did not occur with PCB 153 treatment. 4-Chlorobiphenyl (PCB 3), PCB 28, and PCB 153 treatment of Sprague-Dawley rats resulted in a large increase in protein binding to a consensus activated protein-1 (AP-l) element. However, 3,4-dichlorobiphenyl (PCB 12) and 3,3′,4,4′-tetrachlorobiphenyl (PCB 77) treatments did not increase AP-1 transcription activity. Further analysis of the proteins binding to the AP-1 consensus sequence with antibodies specific for c-fos, junD, and junB indicated that the protein composition consists of junD proteins. These data showed functional differences between noncoplanar and coplanar PCBs with respect to STAT activation and AP-1-DNA binding.

Original languageEnglish (US)
Pages (from-to)199-208
Number of pages10
JournalMolecular Carcinogenesis
Volume30
Issue number4
DOIs
StatePublished - Jan 1 2001

Fingerprint

Polychlorinated Biphenyls
Transcription Factor AP-1
STAT6 Transcription Factor
Luciferases
Transducers
Protein Binding
Liver Extracts
2,4,5,2',4',5'-hexachlorobiphenyl
Electrophoretic Mobility Shift Assay
Sprague Dawley Rats
STAT5 Transcription Factor
Proteins
Antibodies
Consensus Sequence
Hep G2 Cells
DNA-Binding Proteins
Nuclear Proteins
Reporter Genes
Carcinogens
Interferons

Keywords

  • DNA binding
  • Nuclear proteins
  • Tumor promotion

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research

Cite this

Oakley, G. G., Roe, A. L., Blouin, R. A., Twaroski, T. P., Ganguly, T. C., Vore, M., ... Robertson, L. W. (2001). 2,4,4′-Trichlorobiphenyl increases STAT5 transcriptional activity. Molecular Carcinogenesis, 30(4), 199-208. https://doi.org/10.1002/mc.1029

2,4,4′-Trichlorobiphenyl increases STAT5 transcriptional activity. / Oakley, Greg G.; Roe, Amy L.; Blouin, Robert A.; Twaroski, Timothy P.; Ganguly, Tanmoy C.; Vore, Mary; Lehmler, Hans Joachim; Robertson, Larry W.

In: Molecular Carcinogenesis, Vol. 30, No. 4, 01.01.2001, p. 199-208.

Research output: Contribution to journalArticle

Oakley, GG, Roe, AL, Blouin, RA, Twaroski, TP, Ganguly, TC, Vore, M, Lehmler, HJ & Robertson, LW 2001, '2,4,4′-Trichlorobiphenyl increases STAT5 transcriptional activity', Molecular Carcinogenesis, vol. 30, no. 4, pp. 199-208. https://doi.org/10.1002/mc.1029
Oakley GG, Roe AL, Blouin RA, Twaroski TP, Ganguly TC, Vore M et al. 2,4,4′-Trichlorobiphenyl increases STAT5 transcriptional activity. Molecular Carcinogenesis. 2001 Jan 1;30(4):199-208. https://doi.org/10.1002/mc.1029
Oakley, Greg G. ; Roe, Amy L. ; Blouin, Robert A. ; Twaroski, Timothy P. ; Ganguly, Tanmoy C. ; Vore, Mary ; Lehmler, Hans Joachim ; Robertson, Larry W. / 2,4,4′-Trichlorobiphenyl increases STAT5 transcriptional activity. In: Molecular Carcinogenesis. 2001 ; Vol. 30, No. 4. pp. 199-208.
@article{bd265e17d46746c5917a81dd62091fef,
title = "2,4,4′-Trichlorobiphenyl increases STAT5 transcriptional activity",
abstract = "The promoting effects of polychlorinated biphenyls (PCBs) have been studied extensively in a variety of two-stage carcinogenesis models. However, the molecular mechanisms responsible for the promotion effects of PCBs have not been elucidated. We measured the effect of PCBs on DNA-binding proteins involved in cell proliferation and transformation. Male Sprague-Dawley rats were injected intraperitoneally with mono-, di-, tri-, tetra-, or hexachlorobiphenyls (300 μmol/kg/d) each day for 4 d and killed 4 h after the last injection. To detect alterations in nuclear proteins that could explain the tumor-promoter activity of PCBs, liver nuclear extracts were analyzed by electrophoretic mobility shift assays. Electrophoretic mobility shift assay analysis of signal transducers and activators of transcription (STAT)-binding activity to a consensus 7-interferon-activated sequence (GAS) element was compared in liver nuclear extracts from treated rats. STAT-binding activity was eightfold to tenfold higher in nuclear extracts from animals treated with 2,4,4′-trichloro- (PCB 28) and 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB 153). Analysis of the protein complex binding to the GAS element, with antibodies specific for STAT3, STAT5, and STAT6, indicated that the protein complex was made up of STAT5 and STAT6 proteins. HepG2 cells transiently transfected with a luciferase reporter gene construct containing many STAT5 binding sites were treated with PCB 28 and PCB 153. PCB 28 stimulated a greater than 25-fold increase in luciferase activity at the highest concentration tested, 1.0 μg/mL. However, enhanced luciferase activity did not occur with PCB 153 treatment. 4-Chlorobiphenyl (PCB 3), PCB 28, and PCB 153 treatment of Sprague-Dawley rats resulted in a large increase in protein binding to a consensus activated protein-1 (AP-l) element. However, 3,4-dichlorobiphenyl (PCB 12) and 3,3′,4,4′-tetrachlorobiphenyl (PCB 77) treatments did not increase AP-1 transcription activity. Further analysis of the proteins binding to the AP-1 consensus sequence with antibodies specific for c-fos, junD, and junB indicated that the protein composition consists of junD proteins. These data showed functional differences between noncoplanar and coplanar PCBs with respect to STAT activation and AP-1-DNA binding.",
keywords = "DNA binding, Nuclear proteins, Tumor promotion",
author = "Oakley, {Greg G.} and Roe, {Amy L.} and Blouin, {Robert A.} and Twaroski, {Timothy P.} and Ganguly, {Tanmoy C.} and Mary Vore and Lehmler, {Hans Joachim} and Robertson, {Larry W.}",
year = "2001",
month = "1",
day = "1",
doi = "10.1002/mc.1029",
language = "English (US)",
volume = "30",
pages = "199--208",
journal = "Molecular Carcinogenesis",
issn = "0899-1987",
publisher = "Wiley-Liss Inc.",
number = "4",

}

TY - JOUR

T1 - 2,4,4′-Trichlorobiphenyl increases STAT5 transcriptional activity

AU - Oakley, Greg G.

AU - Roe, Amy L.

AU - Blouin, Robert A.

AU - Twaroski, Timothy P.

AU - Ganguly, Tanmoy C.

AU - Vore, Mary

AU - Lehmler, Hans Joachim

AU - Robertson, Larry W.

PY - 2001/1/1

Y1 - 2001/1/1

N2 - The promoting effects of polychlorinated biphenyls (PCBs) have been studied extensively in a variety of two-stage carcinogenesis models. However, the molecular mechanisms responsible for the promotion effects of PCBs have not been elucidated. We measured the effect of PCBs on DNA-binding proteins involved in cell proliferation and transformation. Male Sprague-Dawley rats were injected intraperitoneally with mono-, di-, tri-, tetra-, or hexachlorobiphenyls (300 μmol/kg/d) each day for 4 d and killed 4 h after the last injection. To detect alterations in nuclear proteins that could explain the tumor-promoter activity of PCBs, liver nuclear extracts were analyzed by electrophoretic mobility shift assays. Electrophoretic mobility shift assay analysis of signal transducers and activators of transcription (STAT)-binding activity to a consensus 7-interferon-activated sequence (GAS) element was compared in liver nuclear extracts from treated rats. STAT-binding activity was eightfold to tenfold higher in nuclear extracts from animals treated with 2,4,4′-trichloro- (PCB 28) and 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB 153). Analysis of the protein complex binding to the GAS element, with antibodies specific for STAT3, STAT5, and STAT6, indicated that the protein complex was made up of STAT5 and STAT6 proteins. HepG2 cells transiently transfected with a luciferase reporter gene construct containing many STAT5 binding sites were treated with PCB 28 and PCB 153. PCB 28 stimulated a greater than 25-fold increase in luciferase activity at the highest concentration tested, 1.0 μg/mL. However, enhanced luciferase activity did not occur with PCB 153 treatment. 4-Chlorobiphenyl (PCB 3), PCB 28, and PCB 153 treatment of Sprague-Dawley rats resulted in a large increase in protein binding to a consensus activated protein-1 (AP-l) element. However, 3,4-dichlorobiphenyl (PCB 12) and 3,3′,4,4′-tetrachlorobiphenyl (PCB 77) treatments did not increase AP-1 transcription activity. Further analysis of the proteins binding to the AP-1 consensus sequence with antibodies specific for c-fos, junD, and junB indicated that the protein composition consists of junD proteins. These data showed functional differences between noncoplanar and coplanar PCBs with respect to STAT activation and AP-1-DNA binding.

AB - The promoting effects of polychlorinated biphenyls (PCBs) have been studied extensively in a variety of two-stage carcinogenesis models. However, the molecular mechanisms responsible for the promotion effects of PCBs have not been elucidated. We measured the effect of PCBs on DNA-binding proteins involved in cell proliferation and transformation. Male Sprague-Dawley rats were injected intraperitoneally with mono-, di-, tri-, tetra-, or hexachlorobiphenyls (300 μmol/kg/d) each day for 4 d and killed 4 h after the last injection. To detect alterations in nuclear proteins that could explain the tumor-promoter activity of PCBs, liver nuclear extracts were analyzed by electrophoretic mobility shift assays. Electrophoretic mobility shift assay analysis of signal transducers and activators of transcription (STAT)-binding activity to a consensus 7-interferon-activated sequence (GAS) element was compared in liver nuclear extracts from treated rats. STAT-binding activity was eightfold to tenfold higher in nuclear extracts from animals treated with 2,4,4′-trichloro- (PCB 28) and 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB 153). Analysis of the protein complex binding to the GAS element, with antibodies specific for STAT3, STAT5, and STAT6, indicated that the protein complex was made up of STAT5 and STAT6 proteins. HepG2 cells transiently transfected with a luciferase reporter gene construct containing many STAT5 binding sites were treated with PCB 28 and PCB 153. PCB 28 stimulated a greater than 25-fold increase in luciferase activity at the highest concentration tested, 1.0 μg/mL. However, enhanced luciferase activity did not occur with PCB 153 treatment. 4-Chlorobiphenyl (PCB 3), PCB 28, and PCB 153 treatment of Sprague-Dawley rats resulted in a large increase in protein binding to a consensus activated protein-1 (AP-l) element. However, 3,4-dichlorobiphenyl (PCB 12) and 3,3′,4,4′-tetrachlorobiphenyl (PCB 77) treatments did not increase AP-1 transcription activity. Further analysis of the proteins binding to the AP-1 consensus sequence with antibodies specific for c-fos, junD, and junB indicated that the protein composition consists of junD proteins. These data showed functional differences between noncoplanar and coplanar PCBs with respect to STAT activation and AP-1-DNA binding.

KW - DNA binding

KW - Nuclear proteins

KW - Tumor promotion

UR - http://www.scopus.com/inward/record.url?scp=0035034622&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035034622&partnerID=8YFLogxK

U2 - 10.1002/mc.1029

DO - 10.1002/mc.1029

M3 - Article

C2 - 11346882

AN - SCOPUS:0035034622

VL - 30

SP - 199

EP - 208

JO - Molecular Carcinogenesis

JF - Molecular Carcinogenesis

SN - 0899-1987

IS - 4

ER -