Cholangiocarcinoma(CCA) is enriched of cancer stem cells associated with aggressiveness and drug resistance. FXR, involved in neoplastic transformation of stem cells and/or cholangiocytes, is down-regulated in human CCA cells lines. Our AIM was to evaluate, in primary cultures of human (hiCCA) the effects of the FXR agonist, obeticholic acid (OCA), on the cancerogenic potential of human CCA cells. Primary cell cultures were prepared from mucin- or mixed-hiCCA subtypes from patients. The increasing concentrations (0-5µM) of OCA were added to culture media and, after 3-10 days, their proliferation (MTS assay, population doubling time), apoptosis (AnnexinV-FITC/PI), cell migration and invasion (wound healing and Matrigel® invasion assay) and cancerogenic potential (spheroid formation, clonogenic assay, colony formation capacity) were evaluated. FXR was downregulated (RT-qPCR) in iCCA vs normal hBTSCs (p<0.001) and in mucin-iCCA vs mixed-iCCA (p<0.05). OCA significantly (p<0.05) inhibited proliferation of both mucin-iCCA and mixed-iCCA as low as the concentration of 0.05µM (IC50=0.38µM in mixed-iCCA; 2.1µM in mucin-iCCA). CDCA inhibited cell proliferation, but lower than OCA, consistent with the different potency in FXR activation. The impairment of colony and spheroid formation capacity and delayed wound healing and Matrigel®¬ invasion demonstrated that OCA significantly induced apoptosis of both the subtypes and decreased the in vitro cancerogenic potential of iCCA cells. In general, these effects were more evident against mixed- than mucin-iCCA cell. When administered together with gemcitabine and cisplatin, OCA potentiated the anti-proliferative and pro-apoptotic effects of these chemotherapeutics, but mainly on mixed-iCCA and abolished the capacity of both iCCA-subtypes to form colonies. FXR is down-regulated in iCCA cells, but its activation by OCA results in in vitro anti-cancerogenic effects against both mucin and mixed-iCCA human primary cell cultures. The effects of OCA predominate against mixed-iCCA, consistent with the lower aggressiveness and the higher FXR expression in this CCA subtype.
The FXR Agonist, Obeticholic Acid, Inhibits the Cancerogenic Potential of Primary Human Cholangiocarcinoma Cells: a Study on Primary Human Cell Cultures / Di Matteo, S; Nevi, L; Costantini, D; Colantonio, M; Giulitti, F; Napoletano, C; Safarikia, S; Manzi, E; Derose, Am; Meandro, F; Bragazzi, Mc; Grazi, G; Berloco, Pb; Giuliante, F; Carpino, G; Cardinale, V; Gaudio, E; Alvaro, D. - 50:2(2018), pp. E87-E87. (Intervento presentato al convegno 24th National Congress of Digestive Diseases: Italian Federation of Societies of Digestive Diseases - FISMAD tenutosi a Rome) [10.1016/S1590-8658(18)30310-4].
The FXR Agonist, Obeticholic Acid, Inhibits the Cancerogenic Potential of Primary Human Cholangiocarcinoma Cells: a Study on Primary Human Cell Cultures
Di Matteo, S;Nevi, L;Costantini, D;Giulitti, F;Napoletano, C;Safarikia, S;Bragazzi, MC;Grazi, G;Berloco, PB;Carpino, G;Cardinale, V;Gaudio, E;Alvaro, D
2018
Abstract
Cholangiocarcinoma(CCA) is enriched of cancer stem cells associated with aggressiveness and drug resistance. FXR, involved in neoplastic transformation of stem cells and/or cholangiocytes, is down-regulated in human CCA cells lines. Our AIM was to evaluate, in primary cultures of human (hiCCA) the effects of the FXR agonist, obeticholic acid (OCA), on the cancerogenic potential of human CCA cells. Primary cell cultures were prepared from mucin- or mixed-hiCCA subtypes from patients. The increasing concentrations (0-5µM) of OCA were added to culture media and, after 3-10 days, their proliferation (MTS assay, population doubling time), apoptosis (AnnexinV-FITC/PI), cell migration and invasion (wound healing and Matrigel® invasion assay) and cancerogenic potential (spheroid formation, clonogenic assay, colony formation capacity) were evaluated. FXR was downregulated (RT-qPCR) in iCCA vs normal hBTSCs (p<0.001) and in mucin-iCCA vs mixed-iCCA (p<0.05). OCA significantly (p<0.05) inhibited proliferation of both mucin-iCCA and mixed-iCCA as low as the concentration of 0.05µM (IC50=0.38µM in mixed-iCCA; 2.1µM in mucin-iCCA). CDCA inhibited cell proliferation, but lower than OCA, consistent with the different potency in FXR activation. The impairment of colony and spheroid formation capacity and delayed wound healing and Matrigel®¬ invasion demonstrated that OCA significantly induced apoptosis of both the subtypes and decreased the in vitro cancerogenic potential of iCCA cells. In general, these effects were more evident against mixed- than mucin-iCCA cell. When administered together with gemcitabine and cisplatin, OCA potentiated the anti-proliferative and pro-apoptotic effects of these chemotherapeutics, but mainly on mixed-iCCA and abolished the capacity of both iCCA-subtypes to form colonies. FXR is down-regulated in iCCA cells, but its activation by OCA results in in vitro anti-cancerogenic effects against both mucin and mixed-iCCA human primary cell cultures. The effects of OCA predominate against mixed-iCCA, consistent with the lower aggressiveness and the higher FXR expression in this CCA subtype.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
