TY - JOUR
T1 - Activated hedgehog pathway is a potential target for pharmacological intervention in biliary tract cancer
AU - Kiesslich, Tobias
AU - Mayr, Christian
AU - Wachter, Julia
AU - Bach, Doris
AU - Fuereder, Julia
AU - Wagner, Andrej
AU - Alinger, Beate
AU - Pichler, Martin
AU - Di Fazio, Pietro
AU - Ocker, Matthias
AU - Berr, Frieder
AU - Neureiter, Daniel
N1 - Funding Information:
Acknowledgments This study was supported by funds of the Oes-terreichische Nationalbank (Anniversary fund, project number: 14842), the research fund of the Paracelsus Medical University Salzburg (Grant No. A-12/02/006-KIE) and the ‘Wissenschaftlicher Verein‘of the Institute of Pathology Salzburg. The authors are grateful to the pharmacy at Salzburger Landeskliniken for providing cisplatin.
Publisher Copyright:
© 2014 Springer Science+Business Media New York.
PY - 2014/11/1
Y1 - 2014/11/1
N2 - Hedgehog (Hh) signalling contributes to carcinogenesis and represents a valid druggable target in human cancers, possibly also in biliary tract cancer (BTC). We analysed the expression of Hh components in BTC using eight heterogeneously differentiated cell lines, xenograft tumours and a human tissue microarray. The dose-, time- and cell line-dependent effects of two Hh inhibitors (cyclopamine and Gant-61) were analysed in vitro for survival, apoptosis, cell cycle distribution and possible synergism with conventional chemotherapeutic agents. In human BTC samples, the sonic Hh ligand and the Gli1 transcription factor showed increased expression in tumours compared to normal adjacent tissue and were significantly associated with high tumour grade and positive lymph node status. In BTC cell lines, we could confirm the Hh component expression at varying extent within the employed cell lines in vitro and in vivo indicating non-canonical signalling. Both Hh inhibitors showed dose-dependent cytotoxicity above 5 μM with a stronger effect for Gant-61 inducing apoptosis whereas cyclopamine rather inhibited proliferation. Cytotoxicity was associated with low cytokeratin expression and higher mesenchymal marker expression such as vimentin. Additionally, drug combinations of Gant-61 with conventional chemotherapy (cisplatin) exerted synergistic effects. In conclusion, Hh pathway is significantly activated in human BTC tissue compared to normal adjacent tissue. The current data demonstrate for the first time an effective anticancer activity of especially Gant-61 in BTC and suggest second generation Hh pathway inhibitors as a potential novel treatment strategy in BTC.
AB - Hedgehog (Hh) signalling contributes to carcinogenesis and represents a valid druggable target in human cancers, possibly also in biliary tract cancer (BTC). We analysed the expression of Hh components in BTC using eight heterogeneously differentiated cell lines, xenograft tumours and a human tissue microarray. The dose-, time- and cell line-dependent effects of two Hh inhibitors (cyclopamine and Gant-61) were analysed in vitro for survival, apoptosis, cell cycle distribution and possible synergism with conventional chemotherapeutic agents. In human BTC samples, the sonic Hh ligand and the Gli1 transcription factor showed increased expression in tumours compared to normal adjacent tissue and were significantly associated with high tumour grade and positive lymph node status. In BTC cell lines, we could confirm the Hh component expression at varying extent within the employed cell lines in vitro and in vivo indicating non-canonical signalling. Both Hh inhibitors showed dose-dependent cytotoxicity above 5 μM with a stronger effect for Gant-61 inducing apoptosis whereas cyclopamine rather inhibited proliferation. Cytotoxicity was associated with low cytokeratin expression and higher mesenchymal marker expression such as vimentin. Additionally, drug combinations of Gant-61 with conventional chemotherapy (cisplatin) exerted synergistic effects. In conclusion, Hh pathway is significantly activated in human BTC tissue compared to normal adjacent tissue. The current data demonstrate for the first time an effective anticancer activity of especially Gant-61 in BTC and suggest second generation Hh pathway inhibitors as a potential novel treatment strategy in BTC.
KW - Biliary tract cancer
KW - Cyclopamine
KW - Gant-61
KW - Hedgehog
KW - Oncogenic signalling
KW - Pharmacological inhibition
UR - http://www.scopus.com/inward/record.url?scp=84907592318&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84907592318&partnerID=8YFLogxK
U2 - 10.1007/s11010-014-2161-9
DO - 10.1007/s11010-014-2161-9
M3 - Article
C2 - 25064451
AN - SCOPUS:84907592318
SN - 0300-8177
VL - 396
SP - 257
EP - 268
JO - Molecular and Cellular Biochemistry
JF - Molecular and Cellular Biochemistry
IS - 1-2
ER -