Ganglioside composition correlates with the malignant phenotype of cholangiocarcinoma cells and modulates cell adhesion Background and aims: Gangliosides (GS), a family of sialic acid–containing glycosphingolipids, have been associated with malignant phenotype of several cancers and GD2 has been proposed as a novel CSC-marker. This study aims to characterize the GS profiles of human CCA stem-like subset and their parental cells. Method: Stem-like compartment was enriched by sphere culture (SPH) in two lines of established human intrahepatic CCA cells (HUCCT1, CCLP1). CCA GS compositions were obtained by chromatography. GD2 detection and its co-expression with CD133 CSC-marker was carried out by FACS and immunocytochemical analysis. The role of the GM3 GS was investigated using D-threo-1phenyl-2-palmitoylamino-3-N-morpholine-1-propanol (PPMP), a GM3 synthase inhibitor. Results: In contrast to parental cells grown as adherent monolayers (MON), CCA-SPH showed strong expression of GD2. The time course of SPH-growth revealed a remarkable GD2 expression along with CD133 levels after plating. Immunocytochemistry revealed a heterogeneous GD2-positivity in CCA-SPH. Overall, CCA-SPH profiles showed a GM1/GM2 reduction in parallel with an increase in GM3. Notably, a cell-line specific GS composition was revealed. MON-CCA exhibited high levels of simple GS such as GM3 and GM2 in HUCCT1 and in CCLP1, respectively. Moreover, HUCCT1 showed GM3-dependent adhesion on fibronectin. Remarkably, since GM2 and GD2 biosynthesis is driven by the enzyme beta-1,4 N-acetylgalactosaminyltransferase-1 (B4GALNT1) and GD3 by alpha-Nacetylneuraminide alpha-2,8-sialyltransferase (ST8SIA1), mRNA expression of both enzymes were analyzed by transcriptomic data from surgically resected CCA samples. Both B4GALNT1 and ST8SIA1 were significantly increased in tumor samples compared to paired non-tumoral liver tissue. Strikingly, B4GALNT1 expression considerably correlated with recurrence, perineural invasion and presence of satellite nodules in CCA patients. Conclusion: We show for the first time that the GS composition is modulated in the CCA stem-like subset. GD2 should be explored as a candidate biomarker for CCA. GS composition may affect pivotal characteristics of CCA cells such as adhesion. https://doi.org/10.1016/j.dld.2018.11.049
Ganglioside composition correlates with the malignant phenotype of cholangiocarcinoma cells and modulates cell adhesion / Mannini, A.; Raggi, C.; Correnti, M.; Rovida, E.; Andersen, J.B.; Coulouarn, C.; Marra, F.. - In: DIGESTIVE AND LIVER DISEASE. - ISSN 1590-8658. - ELETTRONICO. - 51:(2019), pp. e7-e7. (Intervento presentato al convegno 52nd Annual Meeting Association for the Study of the Liver - AISF (Associazione Italiana per lo Studio del Fegato) tenutosi a Roma (Italy) nel February, 22, 2019) [10.1016/j.dld.2018.11.049].
Ganglioside composition correlates with the malignant phenotype of cholangiocarcinoma cells and modulates cell adhesion
Mannini, A.
;Raggi, C.
;Rovida, E.
;Marra, F.
2019
Abstract
Ganglioside composition correlates with the malignant phenotype of cholangiocarcinoma cells and modulates cell adhesion Background and aims: Gangliosides (GS), a family of sialic acid–containing glycosphingolipids, have been associated with malignant phenotype of several cancers and GD2 has been proposed as a novel CSC-marker. This study aims to characterize the GS profiles of human CCA stem-like subset and their parental cells. Method: Stem-like compartment was enriched by sphere culture (SPH) in two lines of established human intrahepatic CCA cells (HUCCT1, CCLP1). CCA GS compositions were obtained by chromatography. GD2 detection and its co-expression with CD133 CSC-marker was carried out by FACS and immunocytochemical analysis. The role of the GM3 GS was investigated using D-threo-1phenyl-2-palmitoylamino-3-N-morpholine-1-propanol (PPMP), a GM3 synthase inhibitor. Results: In contrast to parental cells grown as adherent monolayers (MON), CCA-SPH showed strong expression of GD2. The time course of SPH-growth revealed a remarkable GD2 expression along with CD133 levels after plating. Immunocytochemistry revealed a heterogeneous GD2-positivity in CCA-SPH. Overall, CCA-SPH profiles showed a GM1/GM2 reduction in parallel with an increase in GM3. Notably, a cell-line specific GS composition was revealed. MON-CCA exhibited high levels of simple GS such as GM3 and GM2 in HUCCT1 and in CCLP1, respectively. Moreover, HUCCT1 showed GM3-dependent adhesion on fibronectin. Remarkably, since GM2 and GD2 biosynthesis is driven by the enzyme beta-1,4 N-acetylgalactosaminyltransferase-1 (B4GALNT1) and GD3 by alpha-Nacetylneuraminide alpha-2,8-sialyltransferase (ST8SIA1), mRNA expression of both enzymes were analyzed by transcriptomic data from surgically resected CCA samples. Both B4GALNT1 and ST8SIA1 were significantly increased in tumor samples compared to paired non-tumoral liver tissue. Strikingly, B4GALNT1 expression considerably correlated with recurrence, perineural invasion and presence of satellite nodules in CCA patients. Conclusion: We show for the first time that the GS composition is modulated in the CCA stem-like subset. GD2 should be explored as a candidate biomarker for CCA. GS composition may affect pivotal characteristics of CCA cells such as adhesion. https://doi.org/10.1016/j.dld.2018.11.049
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