title: Leukemia inhibitory factor protects cholangiocarcinoma cells from drug-induced apoptosis via a PI3K/AKT-dependent Mcl-1 activation.
abstract: Cholangiocarcinoma is an aggressive, strongly chemoresistant liver malignancy. Leukemia inhibitory factor (LIF), an IL-6 family cytokine, promotes progression of various carcinomas. To investigate the role of LIF in cholangiocarcinoma, we evaluated the expression of LIF and its receptor (LIFR) in human samples. LIF secretion and LIFR expression were assessed in established and primary human cholangiocarcinoma cell lines. In cholangiocarcinoma cells, we tested LIF effects on proliferation, invasion, stem cell-like phenotype, chemotherapy-induced apoptosis (gemcitabine+cisplatin), expression levels of pro-apoptotic (Bax) and anti-apoptotic (Mcl-1) proteins, with/without PI3K inhibition, and of pSTAT3, pERK1/2, pAKT. LIF effect on chemotherapy-induced apoptosis was evaluated after LIFR silencing and Mcl-1 inactivation.Results show that LIF and LIFR expression were higher in neoplastic than in control cholangiocytes; LIF was also expressed by tumor stromal cells. LIF had no effects on cholangiocarcinoma cell proliferation, invasion, and stemness signatures, whilst it counteracted drug-induced apoptosis. Upon LIF stimulation, decreased apoptosis was associated with Mcl-1 and pAKT up-regulation and abolished by PI3K inhibition. LIFR silencing and Mcl-1 blockade restored drug-induced apoptosis.In conclusion, autocrine and paracrine LIF signaling promote chemoresistance in cholangiocarcinoma by up-regulating Mcl-1 via a novel STAT3- and MAPK-independent, PI3K/AKT-dependent pathway. Targeting LIF signaling may increase CCA responsiveness to chemotherapy.
authors: Department of Molecular Medicine, University of Padua, Padua, Italy.
authors: Stuart Duncan
authors: SD
authors: Morton
authors: Department of Molecular Medicine, University of Padua, Padua, Italy.|Department of Surgery & Translational Medicine, University of Milan-Bicocca, Milan, Italy.
authors: Massimiliano
authors: M
authors: Cadamuro
authors: Department of Surgery & Translational Medicine, University of Milan-Bicocca, Milan, Italy.
authors: Simone
authors: S
authors: Brivio
authors: Department of Molecular Medicine, University of Padua, Padua, Italy.|Department of Surgery & Translational Medicine, University of Milan-Bicocca, Milan, Italy.
authors: Marta
authors: M
authors: Vismara
authors: Fourth Surgery Division, Treviso Regional Hospital, Treviso, Italy.
authors: Tommaso
authors: T
authors: Stecca
authors: Fourth Surgery Division, Treviso Regional Hospital, Treviso, Italy.
authors: Marco
authors: M
authors: Massani
authors: Fourth Surgery Division, Treviso Regional Hospital, Treviso, Italy.|Department of Surgical, Oncological and Gastroenterological Sciences, University of Padua, Padua, Italy.
authors: Nicolò
authors: N
authors: Bassi
authors: Pathology Unit, Treviso Regional Hospital, Treviso, Italy.
authors: Alberto
authors: A
authors: Furlanetto
authors: School of Immunity and Infection, University of Birmingham, Birmingham, UK.
authors: Ruth Elizabeth
authors: RE
authors: Joplin
authors: Department of Surgical, Oncological and Gastroenterological Sciences, University of Padua, Padua, Italy.
authors: Annarosa
authors: A
authors: Floreani
authors: Department of Molecular Medicine, University of Padua, Padua, Italy.|Section of Digestive Diseases, Yale University School of Medicine, New Haven, CT, USA.
authors: Luca
authors: L
authors: Fabris
authors: Department of Surgery & Translational Medicine, University of Milan-Bicocca, Milan, Italy.|Section of Digestive Diseases, Yale University School of Medicine, New Haven, CT, USA.
authors: Mario
authors: M
authors: Strazzabosco
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D001650:Bile Duct Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D018281:Cholangiocarcinoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002945:Cisplatin / Q000494:pharmacology; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D053605:Leukemia Inhibitory Factor / Q000235:genetics / Q000378:metabolism*; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D000093542:Gemcitabine
chemical_list: D000970:Antineoplastic Agents; C507837:LIF protein, human; C507840:LIFR protein, human; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; C579095:MCL1 protein, human; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D003841:Deoxycytidine; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D002945:Cisplatin; D000093542:Gemcitabine
keywords: Mcl-1; chemoresistance; cholangiocarcinoma; leukemia inhibitory factor; phosphatidylinositol-3 kinase