Role of the stromal-derived factor-1 (SDF-1)-CXCR4 axis in the interaction between hepatic stellate cells and cholangiocarcinoma.
OBJECTIVE
Cholangiocarcinoma (CCA) is highly fatal because of early invasion, widespread metastasis, and lack of an effective therapy. Migration, invasion, and metastasis of CCA cells are modulated by signals received from stromal cells. The SDF-1-CXCR4 axis emerges as a pivotal regulator of migration and survival of different tumor cells. The aim of the present study was to characterize the interaction between CCA cells and human hepatic stellate cells (hHSC) focusing on the role of SDF-1.


METHODS
The intrahepatic CCA cell line HuCCT-1 and primary hHSC were used for this study. RNA expression was examined by RTQ-PCR and protein expression by Western blotting. Immunofluorescence microscopy and immunohistochemistry were also employed. Migration of CCA cells was assessed using modified Boyden chambers.


RESULTS
CXCR4 was clearly expressed in CCA cells of human CCA liver specimens. SDF-1 and hHSC conditioned medium (CM) promoted HuCCT-1 cell migration, which was abrogated by pre-incubation with AMD3100, a non-peptide antagonist of the CXCR4 receptor. In addition, HuCCT-1 cells silenced for CXCR4 did not migrate in presence of SDF-1. Both P-ERK and p-AKT were implicated in HuCCT-1 migration and showed a biphasic trend under stimulation of SDF-1. Finally, SDF-1 induced apoptotic rescue of HuCCT-1 cells by binding to CXCR4.


CONCLUSIONS
Our study demonstrates that CCA cells migration and survival are modulated by the crosstalk between SDF-1, released by hHSC, and HuCCT-1 cells bearing CXCR4.
Department of Internal Medicine, Center for Research, High Education and Transfer DENOThe, University of Florence, Italy, Largo Brambilla 3, 50134 Florence, Italy. a.gentilini@dmi.unifi.it
Alessandra
A
Gentilini
Krista
K
Rombouts
Sara
S
Galastri
Alessandra
A
Caligiuri
Eleonora
E
Mingarelli
Tommaso
T
Mello
Fabio
F
Marra
Stefano
S
Mantero
Massimo
M
Roncalli
Pietro
P
Invernizzi
Massimo
M
Pinzani
D000328:Adult; D000368:Aged; D017209:Apoptosis / Q000187:drug effects; D001596:Benzylamines; D001650:Bile Duct Neoplasms / Q000378:metabolism*; D001653:Bile Ducts, Intrahepatic; D002450:Cell Communication; D045744:Cell Line, Tumor; D054377:Chemokine CXCL12 / Q000378:metabolism* / Q000494:pharmacology; D002633:Chemotaxis / Q000187:drug effects / Q000235:genetics; D018281:Cholangiocarcinoma / Q000378:metabolism*; D000080027:Cyclams; D005260:Female; D020868:Gene Silencing; D055166:Hepatic Stellate Cells / Q000378:metabolism*; D006571:Heterocyclic Compounds / Q000494:pharmacology; D006801:Humans; D008099:Liver / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008263:Macrophage Migration-Inhibitory Factors / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D061251:Primary Cell Culture; D012313:RNA / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
D001596:Benzylamines; D054377:Chemokine CXCL12; D000080027:Cyclams; D006571:Heterocyclic Compounds; D008263:Macrophage Migration-Inhibitory Factors; D019869:Phosphatidylinositol 3-Kinases; D012313:RNA; D019718:Receptors, CXCR4; C515255:CXCL12 protein, human; C527276:CXCR4 protein, human; C088327:plerixafor