Non-canonical Hedgehog signaling contributes to chemotaxis in cholangiocarcinoma

Nataliya Razumilava, Sergio A. Gradilone, Rory L. Smoot, Joachim C. Mertens, Steven F. Bronk, Alphonse E. Sirica, Gregory James Gores

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Background & Aims The Hedgehog signaling pathway contributes to cholangiocarcinoma biology. However, canonical Hedgehog signaling requires cilia, and cholangiocarcinoma cells often do not express cilia. To resolve this paradox, we examined non-canonical (G-protein coupled, pertussis toxin sensitive) Hedgehog signaling in cholangiocarcinoma cells. Methods Human [non-malignant (H69), malignant (HuCC-T1 and Mz-ChA-1)] and rat [non-malignant (BDE1 and NRC), and malignant (BDEneu)] cell lines were employed for this study. A BDEΔLoop2 cell line with the dominant-negative receptor Patched-1 was generated with the Sleeping Beauty transposon transfection system. Results Cilia expression was readily identified in non-malignant, but not in malignant cholangiocarcinoma cell lines. Although the canonical Hh signaling pathway was markedly attenuated in cholangiocarcinoma cells, they were chemotactic to purmorphamine, a small-molecule direct Smoothened agonist. Purmorphamine also induced remodeling of the actin cytoskeleton with formation of filopodia and lamellipodia-like protrusions. All these biological features of cell migration were pertussis toxin sensitive, a feature of G-protein coupled (Gis) receptors. To further test the role of Hedgehog signaling in vivo, we employed a syngeneic orthotopic rat model of cholangiocarcinoma. In vivo, genetic inhibition of the Hedgehog signaling pathway employing BDE ΔLoop2 cells or pharmacological inhibition with a small-molecule antagonist of Smoothened, vismodegib, was tumor and metastasis suppressive. Conclusions Cholangiocarcinoma cells exhibit non-canonical Hedgehog signaling with chemotaxis despite impaired cilia expression. This non-canonical Hedgehog signaling pathway appears to contribute to cholangiocarcinoma progression, thereby, supporting a role for Hedgehog pathway inhibition in human cholangiocarcinoma.

Original languageEnglish (US)
Pages (from-to)599-605
Number of pages7
JournalJournal of Hepatology
Volume60
Issue number3
DOIs
StatePublished - Mar 2014

Fingerprint

Cholangiocarcinoma
Hedgehogs
Chemotaxis
Cilia
Pseudopodia
Pertussis Toxin
HhAntag691
Cell Line
Beauty
G-Protein-Coupled Receptors
Actin Cytoskeleton
GTP-Binding Proteins
Cell Movement
Transfection
Pharmacology
Neoplasm Metastasis

Keywords

  • Biliary tract cancer
  • Dominant-negative Ptch1
  • G-protein coupled receptor
  • Patched-1
  • Smoothened

ASJC Scopus subject areas

  • Hepatology
  • Medicine(all)

Cite this

Razumilava, N., Gradilone, S. A., Smoot, R. L., Mertens, J. C., Bronk, S. F., Sirica, A. E., & Gores, G. J. (2014). Non-canonical Hedgehog signaling contributes to chemotaxis in cholangiocarcinoma. Journal of Hepatology, 60(3), 599-605. https://doi.org/10.1016/j.jhep.2013.11.005

Non-canonical Hedgehog signaling contributes to chemotaxis in cholangiocarcinoma. / Razumilava, Nataliya; Gradilone, Sergio A.; Smoot, Rory L.; Mertens, Joachim C.; Bronk, Steven F.; Sirica, Alphonse E.; Gores, Gregory James.

In: Journal of Hepatology, Vol. 60, No. 3, 03.2014, p. 599-605.

Research output: Contribution to journalArticle

Razumilava, N, Gradilone, SA, Smoot, RL, Mertens, JC, Bronk, SF, Sirica, AE & Gores, GJ 2014, 'Non-canonical Hedgehog signaling contributes to chemotaxis in cholangiocarcinoma', Journal of Hepatology, vol. 60, no. 3, pp. 599-605. https://doi.org/10.1016/j.jhep.2013.11.005
Razumilava N, Gradilone SA, Smoot RL, Mertens JC, Bronk SF, Sirica AE et al. Non-canonical Hedgehog signaling contributes to chemotaxis in cholangiocarcinoma. Journal of Hepatology. 2014 Mar;60(3):599-605. https://doi.org/10.1016/j.jhep.2013.11.005
Razumilava, Nataliya ; Gradilone, Sergio A. ; Smoot, Rory L. ; Mertens, Joachim C. ; Bronk, Steven F. ; Sirica, Alphonse E. ; Gores, Gregory James. / Non-canonical Hedgehog signaling contributes to chemotaxis in cholangiocarcinoma. In: Journal of Hepatology. 2014 ; Vol. 60, No. 3. pp. 599-605.
@article{033cc153316940e0bb17ef9a2cc49f20,
title = "Non-canonical Hedgehog signaling contributes to chemotaxis in cholangiocarcinoma",
abstract = "Background & Aims The Hedgehog signaling pathway contributes to cholangiocarcinoma biology. However, canonical Hedgehog signaling requires cilia, and cholangiocarcinoma cells often do not express cilia. To resolve this paradox, we examined non-canonical (G-protein coupled, pertussis toxin sensitive) Hedgehog signaling in cholangiocarcinoma cells. Methods Human [non-malignant (H69), malignant (HuCC-T1 and Mz-ChA-1)] and rat [non-malignant (BDE1 and NRC), and malignant (BDEneu)] cell lines were employed for this study. A BDEΔLoop2 cell line with the dominant-negative receptor Patched-1 was generated with the Sleeping Beauty transposon transfection system. Results Cilia expression was readily identified in non-malignant, but not in malignant cholangiocarcinoma cell lines. Although the canonical Hh signaling pathway was markedly attenuated in cholangiocarcinoma cells, they were chemotactic to purmorphamine, a small-molecule direct Smoothened agonist. Purmorphamine also induced remodeling of the actin cytoskeleton with formation of filopodia and lamellipodia-like protrusions. All these biological features of cell migration were pertussis toxin sensitive, a feature of G-protein coupled (Gis) receptors. To further test the role of Hedgehog signaling in vivo, we employed a syngeneic orthotopic rat model of cholangiocarcinoma. In vivo, genetic inhibition of the Hedgehog signaling pathway employing BDE ΔLoop2 cells or pharmacological inhibition with a small-molecule antagonist of Smoothened, vismodegib, was tumor and metastasis suppressive. Conclusions Cholangiocarcinoma cells exhibit non-canonical Hedgehog signaling with chemotaxis despite impaired cilia expression. This non-canonical Hedgehog signaling pathway appears to contribute to cholangiocarcinoma progression, thereby, supporting a role for Hedgehog pathway inhibition in human cholangiocarcinoma.",
keywords = "Biliary tract cancer, Dominant-negative Ptch1, G-protein coupled receptor, Patched-1, Smoothened",
author = "Nataliya Razumilava and Gradilone, {Sergio A.} and Smoot, {Rory L.} and Mertens, {Joachim C.} and Bronk, {Steven F.} and Sirica, {Alphonse E.} and Gores, {Gregory James}",
year = "2014",
month = "3",
doi = "10.1016/j.jhep.2013.11.005",
language = "English (US)",
volume = "60",
pages = "599--605",
journal = "Journal of Hepatology",
issn = "0168-8278",
publisher = "Elsevier",
number = "3",

}

TY - JOUR

T1 - Non-canonical Hedgehog signaling contributes to chemotaxis in cholangiocarcinoma

AU - Razumilava, Nataliya

AU - Gradilone, Sergio A.

AU - Smoot, Rory L.

AU - Mertens, Joachim C.

AU - Bronk, Steven F.

AU - Sirica, Alphonse E.

AU - Gores, Gregory James

PY - 2014/3

Y1 - 2014/3

N2 - Background & Aims The Hedgehog signaling pathway contributes to cholangiocarcinoma biology. However, canonical Hedgehog signaling requires cilia, and cholangiocarcinoma cells often do not express cilia. To resolve this paradox, we examined non-canonical (G-protein coupled, pertussis toxin sensitive) Hedgehog signaling in cholangiocarcinoma cells. Methods Human [non-malignant (H69), malignant (HuCC-T1 and Mz-ChA-1)] and rat [non-malignant (BDE1 and NRC), and malignant (BDEneu)] cell lines were employed for this study. A BDEΔLoop2 cell line with the dominant-negative receptor Patched-1 was generated with the Sleeping Beauty transposon transfection system. Results Cilia expression was readily identified in non-malignant, but not in malignant cholangiocarcinoma cell lines. Although the canonical Hh signaling pathway was markedly attenuated in cholangiocarcinoma cells, they were chemotactic to purmorphamine, a small-molecule direct Smoothened agonist. Purmorphamine also induced remodeling of the actin cytoskeleton with formation of filopodia and lamellipodia-like protrusions. All these biological features of cell migration were pertussis toxin sensitive, a feature of G-protein coupled (Gis) receptors. To further test the role of Hedgehog signaling in vivo, we employed a syngeneic orthotopic rat model of cholangiocarcinoma. In vivo, genetic inhibition of the Hedgehog signaling pathway employing BDE ΔLoop2 cells or pharmacological inhibition with a small-molecule antagonist of Smoothened, vismodegib, was tumor and metastasis suppressive. Conclusions Cholangiocarcinoma cells exhibit non-canonical Hedgehog signaling with chemotaxis despite impaired cilia expression. This non-canonical Hedgehog signaling pathway appears to contribute to cholangiocarcinoma progression, thereby, supporting a role for Hedgehog pathway inhibition in human cholangiocarcinoma.

AB - Background & Aims The Hedgehog signaling pathway contributes to cholangiocarcinoma biology. However, canonical Hedgehog signaling requires cilia, and cholangiocarcinoma cells often do not express cilia. To resolve this paradox, we examined non-canonical (G-protein coupled, pertussis toxin sensitive) Hedgehog signaling in cholangiocarcinoma cells. Methods Human [non-malignant (H69), malignant (HuCC-T1 and Mz-ChA-1)] and rat [non-malignant (BDE1 and NRC), and malignant (BDEneu)] cell lines were employed for this study. A BDEΔLoop2 cell line with the dominant-negative receptor Patched-1 was generated with the Sleeping Beauty transposon transfection system. Results Cilia expression was readily identified in non-malignant, but not in malignant cholangiocarcinoma cell lines. Although the canonical Hh signaling pathway was markedly attenuated in cholangiocarcinoma cells, they were chemotactic to purmorphamine, a small-molecule direct Smoothened agonist. Purmorphamine also induced remodeling of the actin cytoskeleton with formation of filopodia and lamellipodia-like protrusions. All these biological features of cell migration were pertussis toxin sensitive, a feature of G-protein coupled (Gis) receptors. To further test the role of Hedgehog signaling in vivo, we employed a syngeneic orthotopic rat model of cholangiocarcinoma. In vivo, genetic inhibition of the Hedgehog signaling pathway employing BDE ΔLoop2 cells or pharmacological inhibition with a small-molecule antagonist of Smoothened, vismodegib, was tumor and metastasis suppressive. Conclusions Cholangiocarcinoma cells exhibit non-canonical Hedgehog signaling with chemotaxis despite impaired cilia expression. This non-canonical Hedgehog signaling pathway appears to contribute to cholangiocarcinoma progression, thereby, supporting a role for Hedgehog pathway inhibition in human cholangiocarcinoma.

KW - Biliary tract cancer

KW - Dominant-negative Ptch1

KW - G-protein coupled receptor

KW - Patched-1

KW - Smoothened

UR - http://www.scopus.com/inward/record.url?scp=84894106141&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84894106141&partnerID=8YFLogxK

U2 - 10.1016/j.jhep.2013.11.005

DO - 10.1016/j.jhep.2013.11.005

M3 - Article

C2 - 24239776

AN - SCOPUS:84894106141

VL - 60

SP - 599

EP - 605

JO - Journal of Hepatology

JF - Journal of Hepatology

SN - 0168-8278

IS - 3

ER -