Cholest-4,6-Dien-3-One Promote Epithelial-To-Mesenchymal Transition (EMT) in Biliary Tree Stem/Progenitor Cell Cultures In Vitro

Abstract

Human biliary tree stem/progenitor cells (hBTSCs), reside in peribiliary glands, are mainly stimulated by primary sclerosing cholangitis (PSC) and cholangiocarcinoma. In these pathologies, hBTSCs displayed epithelial-to-mesenchymal transition (EMT), senescence characteristics, and impaired differentiation. Here, we investigated the effects of cholest-4,6-dien-3-one, an oxysterol involved in cholangiopathies, on hBTSCs biology. hBTSCs were isolated from donor organs, cultured in self-renewal control conditions, differentiated in mature cholangiocytes by specifically tailored medium, or exposed for 10 days to concentration of cholest-4,6-dien-3-one (0.14 mM). Viability, proliferation, senescence, EMT genes expression, telomerase activity, interleukin 6 (IL6) secretion, differentiation capacity, and HDAC6 gene expression were analyzed. Although the effect of cholest-4,6-dien-3-one was not detected on hBTSCs viability, we found a significant increase in cell proliferation, senescence, and IL6 secretion. Interestingly, cholest-4.6-dien-3-one impaired differentiation in mature cholangiocytes and, simultaneously, induced the EMT markers, significantly reduced the telomerase activity, and induced HDAC6 gene expression. Moreover, cholest-4,6-dien-3-one enhanced bone morphogenic protein 4 (Bmp-4) and sonic hedgehog (Shh) pathways in hBTSCs. The same pathways activated by human recombinant proteins induced the expression of EMT markers in hBTSCs. In conclusion, we demonstrated that chronic exposition of cholest-4,6-dien-3-one induced cell proliferation, EMT markers, and senescence in hBTSC, and also impaired the differentiation in mature cholangiocytes.

The protease‐inhibitor SerpinB3 as a critical modulator of the stem‐like subset in human cholangiocarcinoma

“…indicating CSCs as a driving force of CCA initiation, dissemination and drug-resistance. [4][5][6][7][8][9][10][11][12][13][14][15][16] SerpinB3 (formerly known as squamous cell carcinoma antigen-1 or SCCA1) is a member of the serine-protease inhibitors family. 17 It is expressed in normal epithelial cells and overexpressed in cancer cells and damaged hepatocytes.…”

“…CSCs, also referred to as tumour‐initiating cells or tumour‐propagating cells, are tumorigenic, metastatic, resistant to chemo‐ and radiation therapy and are responsible for tumour recurrence. Recently, we and others have provided data indicating CSCs as a driving force of CCA initiation, dissemination and drug‐resistance 4‐16 …”

“…Recently, we and others have provided data indicating CSCs as a driving force of CCA initiation, dissemination and drug‐resistance. 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 …”

Impact of Oxysterols on Cell Death, Proliferation, and Differentiation Induction: Current Status

“…7β,27-dihydroxycholesterol and 7α,27-dihydroxycholesterol are RAR-related orphan receptor gamma t (RORγt) ligands in both murine and human primary CD4+ T, promoting differentiation of these cells into IL-17–producing CD4+ Th17 T cells in vitro [ 121 ]. Cholest-4,6-dien-3-one induces epithelial-to-mesenchymal transition on human biliary tree stem/progenitor cells, while it impairs the differentiation of these cells in mature cholangiocytes and reduces their telomerase activity [ 110 ].…”

Hepatocyte Smoothened Activity Controls Susceptibility to Insulin Resistance and Nonalcoholic Fatty Liver Disease

“…The Hedgehog pathway is a proven regulator of state transitions in many of the cells that participate in hepatic wound healing responses, including hepatic stellate cells, immune cells, sinusoidal endothelial cells and bile duct cells (cholangiocytes). 8 , 9 , 10 , 11 Each of these cell types not only responds to Hedgehog ligands, but also can produce them; thus, Hedgehog-sensitive autocrine and paracrine interactions are central to liver repair. 12 , 13 , 14 …”