Background and Aim Intrahepatic cholangiocarcinoma (iCCA) represents a rare and invasive cancer that develops in the biliary system and bears a fatal prognosis (5-year relative survival rate). At present, the issue is a worldwide health concern that demands immediate action. The World Health Organization (ICD-O-3.2) has approved the classification of iCCA into small and large bile duct types according to their molecular, histological, and clinical features. The Hedgehog (Hh) pathway is essential in the development and progression of iCCA. Certainly, it is essential in multiple iCCA characteristics like tumor development, survival, activation of cancer stem cells, and epithelial-mesenchymal transition. This study aims to evaluate how Glabrescione B (GlaB), a novel natural compound, impacts Gli1 inhibition in primary and established cell lines in vitro, focusing on the key transcriptional factor and ultimate effector in the Hh pathway. Materials and Methods The Trypan Blue Exclusion test was used to evaluate the reactivity of free GlaB and GlaB encapsulated in hyaluronic acid (HA-GlaB) nanoparticles. MTS assay was performed to determine the IC50 value. Western blot has been used to analyze the expression levels of the protein target. Wound-healing assay assesses cells' migration capability. A colony formation assay was performed to evaluate the capacity of cancer cells to generate colonies. Flow cytometry has been used to analyze cell death, including apoptosis and necrosis. The results were confirmed through at least three different experiments and the quantitative data was displayed as the mean ± SEM. The Student's t-test was used to compare differences in variances between two groups in related samples. A p-value below 0.05 was considered statistically significant (*p<0.05; **p<0.01; ***p<0.001). Results Our findings indicate that when iCCA cells are treated with either free GlaB or HA-GlaB, there is a notable reduction in cell growth, survival, migration, ability to form colonies, and levels of Gli1 protein in a manner that depends on the dosage and duration of treatment (0.05<0.001). Moreover, the drug is able to induce iCCA cell death after the administration, compared to the controls. Conclusions In conclusion, the new natural compound Glabrescione B can inhibit the growth, survival, invasiveness of iCCA cells, and induce their death. Together, this data provides insights into a new and potentially beneficial natural chemical compound for the treatment of iCCA in vivo.
Deciphering the anticancer activity of Glabrescione B in Intrahepatic Cholangiocarcinoma / Paradiso, S.; Carpino, G.; Di Marcotullio, L.; Infante, P.; Di Meo, C.; Quaglio, D.; Franchitto, M.; De Luca, T.; Botta, B.; Gaudio, E.; Alvaro, D.; Cardinale, V.. - (2025). (Intervento presentato al convegno 31° CONGRESSO NAZIONALE DELLE MALATTIE DIGESTIVE - FISMAD tenutosi a Rome, Italy).
Deciphering the anticancer activity of Glabrescione B in Intrahepatic Cholangiocarcinoma
Paradiso S.
Primo
Investigation
;Carpino G.;Di Marcotullio L.;Infante P.;Di Meo C.;Quaglio D.;Franchitto M.;De Luca T.;Botta B.;Gaudio E.Funding Acquisition
;Alvaro D.Penultimo
Funding Acquisition
;Cardinale V.
Ultimo
Supervision
2025
Abstract
Background and Aim Intrahepatic cholangiocarcinoma (iCCA) represents a rare and invasive cancer that develops in the biliary system and bears a fatal prognosis (5-year relative survival rate). At present, the issue is a worldwide health concern that demands immediate action. The World Health Organization (ICD-O-3.2) has approved the classification of iCCA into small and large bile duct types according to their molecular, histological, and clinical features. The Hedgehog (Hh) pathway is essential in the development and progression of iCCA. Certainly, it is essential in multiple iCCA characteristics like tumor development, survival, activation of cancer stem cells, and epithelial-mesenchymal transition. This study aims to evaluate how Glabrescione B (GlaB), a novel natural compound, impacts Gli1 inhibition in primary and established cell lines in vitro, focusing on the key transcriptional factor and ultimate effector in the Hh pathway. Materials and Methods The Trypan Blue Exclusion test was used to evaluate the reactivity of free GlaB and GlaB encapsulated in hyaluronic acid (HA-GlaB) nanoparticles. MTS assay was performed to determine the IC50 value. Western blot has been used to analyze the expression levels of the protein target. Wound-healing assay assesses cells' migration capability. A colony formation assay was performed to evaluate the capacity of cancer cells to generate colonies. Flow cytometry has been used to analyze cell death, including apoptosis and necrosis. The results were confirmed through at least three different experiments and the quantitative data was displayed as the mean ± SEM. The Student's t-test was used to compare differences in variances between two groups in related samples. A p-value below 0.05 was considered statistically significant (*p<0.05; **p<0.01; ***p<0.001). Results Our findings indicate that when iCCA cells are treated with either free GlaB or HA-GlaB, there is a notable reduction in cell growth, survival, migration, ability to form colonies, and levels of Gli1 protein in a manner that depends on the dosage and duration of treatment (0.05<0.001). Moreover, the drug is able to induce iCCA cell death after the administration, compared to the controls. Conclusions In conclusion, the new natural compound Glabrescione B can inhibit the growth, survival, invasiveness of iCCA cells, and induce their death. Together, this data provides insights into a new and potentially beneficial natural chemical compound for the treatment of iCCA in vivo.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
