Ethanol-TGFα-MEK Signaling Promotes Growth of Human Hepatocellular Carcinoma

Matthew Hennig, Michele Yip-Schneider, Patrick Klein, Sabrina Wentz, Jesus M. Matos, Courtney Doyle, Jennifer Choi, Huangbing Wu, Amanda O'Mara, Alex Menze, Stephen Noble, Iain H. McKillop, C. Schmidt

Research output: Contribution to journalArticle

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Abstract

Background: Chronic ethanol intake is a significant risk factor for the development of cirrhosis and hepatocellular carcinoma (HCC). The effects of ethanol on extracellular signal-regulated kinase (ERK) activation, transforming growth factor alpha (TGF-α), and HCC growth were examined in this study. Methods: HepG2, SKHep, Hep3B human HCC cells, or normal human hepatocytes were treated with ethanol (0-100 mM), exogenous TGF-α, TGF-α neutralization antibody or the MEK inhibitor U0126. TGF-α levels were quantified by ELISA. Growth was determined by trypan blue-excluded cell counts. Cell cycle phase distribution was determined by flow cytometry. Protein expression was determined by Western blot. Results: Ethanol treatment (10-40 mM) increased ERK activation in HepG2 and SKHep HCC cells but not in Hep3B or human hepatocyte cells. Growth increased in HepG2 (174 ± 29%, P < 0.05) and SKHep (149 ± 12%, P < 0.05) cells in response to ethanol treatment. Correspondingly, ethanol increased S phase distribution in these cells. U0126 suppressed ethanol-induced growth increases. Ethanol treatment for 24 h also raised TGF-α levels in HepG2 cells (118%-198%) and SKHep cells (112%-177%). Exogenous administration of recombinant TGF-α mimicked the ethanol-induced growth in HepG2 and SKHep cells; TGF-α neutralization antibody effectively abrogated this effect. The TGF-a neutralization antibody also prevented ERK activation by ethanol in HepG2 cells. Conclusions: These data demonstrate that clinically relevant doses of ethanol stimulate ERK-dependent proliferation of HCC cells. Ethanol up-regulates TGF-α levels in HCC cells and enhances growth through cell cycles changes, which appear to be mediated through TGF-α-MEK-ERK signaling. Ethanol-MEK signaling in normal hepatocytes is absent, suggesting that ethanol promotion of HCC growth may in part depend upon the acquisition of cancer-specific signaling by hepatocytes.

Original languageEnglish
Pages (from-to)187-195
Number of pages9
JournalJournal of Surgical Research
Volume154
Issue number2
DOIs
StatePublished - Jun 15 2009

Fingerprint

Transforming Growth Factor alpha
Mitogen-Activated Protein Kinase Kinases
Hepatocellular Carcinoma
Ethanol
Growth
Extracellular Signal-Regulated MAP Kinases
Hepatocytes
Hep G2 Cells
Antibodies
Cell Cycle
Trypan Blue
S Phase

Keywords

  • ERK
  • ethanol
  • HCC
  • hepatocellular carcinoma
  • MEK
  • TGF-α

ASJC Scopus subject areas

  • Surgery
  • Medicine(all)

Cite this

Ethanol-TGFα-MEK Signaling Promotes Growth of Human Hepatocellular Carcinoma. / Hennig, Matthew; Yip-Schneider, Michele; Klein, Patrick; Wentz, Sabrina; Matos, Jesus M.; Doyle, Courtney; Choi, Jennifer; Wu, Huangbing; O'Mara, Amanda; Menze, Alex; Noble, Stephen; McKillop, Iain H.; Schmidt, C.

In: Journal of Surgical Research, Vol. 154, No. 2, 15.06.2009, p. 187-195.

Research output: Contribution to journalArticle

Hennig, M, Yip-Schneider, M, Klein, P, Wentz, S, Matos, JM, Doyle, C, Choi, J, Wu, H, O'Mara, A, Menze, A, Noble, S, McKillop, IH & Schmidt, C 2009, 'Ethanol-TGFα-MEK Signaling Promotes Growth of Human Hepatocellular Carcinoma', Journal of Surgical Research, vol. 154, no. 2, pp. 187-195. https://doi.org/10.1016/j.jss.2008.11.836
Hennig, Matthew ; Yip-Schneider, Michele ; Klein, Patrick ; Wentz, Sabrina ; Matos, Jesus M. ; Doyle, Courtney ; Choi, Jennifer ; Wu, Huangbing ; O'Mara, Amanda ; Menze, Alex ; Noble, Stephen ; McKillop, Iain H. ; Schmidt, C. / Ethanol-TGFα-MEK Signaling Promotes Growth of Human Hepatocellular Carcinoma. In: Journal of Surgical Research. 2009 ; Vol. 154, No. 2. pp. 187-195.
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abstract = "Background: Chronic ethanol intake is a significant risk factor for the development of cirrhosis and hepatocellular carcinoma (HCC). The effects of ethanol on extracellular signal-regulated kinase (ERK) activation, transforming growth factor alpha (TGF-α), and HCC growth were examined in this study. Methods: HepG2, SKHep, Hep3B human HCC cells, or normal human hepatocytes were treated with ethanol (0-100 mM), exogenous TGF-α, TGF-α neutralization antibody or the MEK inhibitor U0126. TGF-α levels were quantified by ELISA. Growth was determined by trypan blue-excluded cell counts. Cell cycle phase distribution was determined by flow cytometry. Protein expression was determined by Western blot. Results: Ethanol treatment (10-40 mM) increased ERK activation in HepG2 and SKHep HCC cells but not in Hep3B or human hepatocyte cells. Growth increased in HepG2 (174 ± 29{\%}, P < 0.05) and SKHep (149 ± 12{\%}, P < 0.05) cells in response to ethanol treatment. Correspondingly, ethanol increased S phase distribution in these cells. U0126 suppressed ethanol-induced growth increases. Ethanol treatment for 24 h also raised TGF-α levels in HepG2 cells (118{\%}-198{\%}) and SKHep cells (112{\%}-177{\%}). Exogenous administration of recombinant TGF-α mimicked the ethanol-induced growth in HepG2 and SKHep cells; TGF-α neutralization antibody effectively abrogated this effect. The TGF-a neutralization antibody also prevented ERK activation by ethanol in HepG2 cells. Conclusions: These data demonstrate that clinically relevant doses of ethanol stimulate ERK-dependent proliferation of HCC cells. Ethanol up-regulates TGF-α levels in HCC cells and enhances growth through cell cycles changes, which appear to be mediated through TGF-α-MEK-ERK signaling. Ethanol-MEK signaling in normal hepatocytes is absent, suggesting that ethanol promotion of HCC growth may in part depend upon the acquisition of cancer-specific signaling by hepatocytes.",
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AU - Wentz, Sabrina

AU - Matos, Jesus M.

AU - Doyle, Courtney

AU - Choi, Jennifer

AU - Wu, Huangbing

AU - O'Mara, Amanda

AU - Menze, Alex

AU - Noble, Stephen

AU - McKillop, Iain H.

AU - Schmidt, C.

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N2 - Background: Chronic ethanol intake is a significant risk factor for the development of cirrhosis and hepatocellular carcinoma (HCC). The effects of ethanol on extracellular signal-regulated kinase (ERK) activation, transforming growth factor alpha (TGF-α), and HCC growth were examined in this study. Methods: HepG2, SKHep, Hep3B human HCC cells, or normal human hepatocytes were treated with ethanol (0-100 mM), exogenous TGF-α, TGF-α neutralization antibody or the MEK inhibitor U0126. TGF-α levels were quantified by ELISA. Growth was determined by trypan blue-excluded cell counts. Cell cycle phase distribution was determined by flow cytometry. Protein expression was determined by Western blot. Results: Ethanol treatment (10-40 mM) increased ERK activation in HepG2 and SKHep HCC cells but not in Hep3B or human hepatocyte cells. Growth increased in HepG2 (174 ± 29%, P < 0.05) and SKHep (149 ± 12%, P < 0.05) cells in response to ethanol treatment. Correspondingly, ethanol increased S phase distribution in these cells. U0126 suppressed ethanol-induced growth increases. Ethanol treatment for 24 h also raised TGF-α levels in HepG2 cells (118%-198%) and SKHep cells (112%-177%). Exogenous administration of recombinant TGF-α mimicked the ethanol-induced growth in HepG2 and SKHep cells; TGF-α neutralization antibody effectively abrogated this effect. The TGF-a neutralization antibody also prevented ERK activation by ethanol in HepG2 cells. Conclusions: These data demonstrate that clinically relevant doses of ethanol stimulate ERK-dependent proliferation of HCC cells. Ethanol up-regulates TGF-α levels in HCC cells and enhances growth through cell cycles changes, which appear to be mediated through TGF-α-MEK-ERK signaling. Ethanol-MEK signaling in normal hepatocytes is absent, suggesting that ethanol promotion of HCC growth may in part depend upon the acquisition of cancer-specific signaling by hepatocytes.

AB - Background: Chronic ethanol intake is a significant risk factor for the development of cirrhosis and hepatocellular carcinoma (HCC). The effects of ethanol on extracellular signal-regulated kinase (ERK) activation, transforming growth factor alpha (TGF-α), and HCC growth were examined in this study. Methods: HepG2, SKHep, Hep3B human HCC cells, or normal human hepatocytes were treated with ethanol (0-100 mM), exogenous TGF-α, TGF-α neutralization antibody or the MEK inhibitor U0126. TGF-α levels were quantified by ELISA. Growth was determined by trypan blue-excluded cell counts. Cell cycle phase distribution was determined by flow cytometry. Protein expression was determined by Western blot. Results: Ethanol treatment (10-40 mM) increased ERK activation in HepG2 and SKHep HCC cells but not in Hep3B or human hepatocyte cells. Growth increased in HepG2 (174 ± 29%, P < 0.05) and SKHep (149 ± 12%, P < 0.05) cells in response to ethanol treatment. Correspondingly, ethanol increased S phase distribution in these cells. U0126 suppressed ethanol-induced growth increases. Ethanol treatment for 24 h also raised TGF-α levels in HepG2 cells (118%-198%) and SKHep cells (112%-177%). Exogenous administration of recombinant TGF-α mimicked the ethanol-induced growth in HepG2 and SKHep cells; TGF-α neutralization antibody effectively abrogated this effect. The TGF-a neutralization antibody also prevented ERK activation by ethanol in HepG2 cells. Conclusions: These data demonstrate that clinically relevant doses of ethanol stimulate ERK-dependent proliferation of HCC cells. Ethanol up-regulates TGF-α levels in HCC cells and enhances growth through cell cycles changes, which appear to be mediated through TGF-α-MEK-ERK signaling. Ethanol-MEK signaling in normal hepatocytes is absent, suggesting that ethanol promotion of HCC growth may in part depend upon the acquisition of cancer-specific signaling by hepatocytes.

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