Journal: Cancers
Article Title: Niclosamide and Palbociclib Act Synergistically to Reduce Cholangiocarcinoma Cell Viability In Vitro and Inhibit Tumour Growth in a Mouse Model
doi: 10.3390/cancers17223721
Figure Lengend Snippet: Drug screening and validation of Niclosamide using CCA cell lines and primary biliary epithelial cells. ( A ) A library of 104 off-patent drugs was screened using the CCLP CCA cell line. CCLP cells were plated at 1 × 10 4 cells per well in a 96 well plate and treated with each drug at its clinically relevant peak serum concentration for 96 h. Cell viability was then determined using an MTT assay and normalised to cells treated with the corresponding vehicle control for each drug. N = 3 biological repeats with a paired, two-tailed T-test followed by Benjamini–Hochberg multiple corrections. The inhibition of viability (%) is plotted against the reciprocal of the p value for each drug and Niclosamide is labelled. ( B – E ) Niclosamide dose–response curves for CCA cell lines (CCLP, RBE, and KKU-M055) and primary biliary epithelial cells (BECs) plated as in ( A ) and treated with Niclosamide [10 nM–100 μM] or vehicle control for 72 h. Cell viability was measured using an MTT assay and normalised to vehicle control. N = 3 biological repeats each performed in triplicate. Error bars represent SEM. When error bars cannot be seen they are smaller than the symbols. ( F ) The graph shows the relative EC50 values for each cell type calculated using GraphPad Prism. Error bars represent SEM (* = p < 0.05 unpaired t -test). The inserted Western blot shows PRH levels in the CCA cell lines prior to treatment and with β-actin as a loading control.
Article Snippet: Primary Biliary Epithelial Cells (BECs) obtained from healthy human liver tissue (Innoprot, Elexalde Derio, Spain) were grown in Epithelial Cell Media (Innoprot, P60106 , Elexalde Derio, Spain) supplemented with 2% FBS and 1% Epithelial Growth Supplement, as per the supplier’s protocol for a maximum of ten passages.
Techniques: Drug discovery, Biomarker Discovery, Concentration Assay, MTT Assay, Control, Two Tailed Test, Inhibition, Western Blot
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![Drug screening and validation of Niclosamide using CCA cell lines and primary biliary <t>epithelial</t> cells. ( A ) A library of 104 off-patent drugs was screened using the CCLP CCA cell line. CCLP cells were plated at 1 × 10 4 cells per well in a 96 well plate and treated with each drug at its clinically relevant peak serum concentration for 96 h. Cell viability was then determined using an MTT assay and normalised to cells treated with the corresponding vehicle control for each drug. N = 3 biological repeats with a paired, two-tailed T-test followed by Benjamini–Hochberg multiple corrections. The inhibition of viability (%) is plotted against the reciprocal of the p value for each drug and Niclosamide is labelled. ( B – E ) Niclosamide dose–response curves for CCA cell lines (CCLP, RBE, and KKU-M055) and primary biliary epithelial cells (BECs) plated as in ( A ) and treated with Niclosamide [10 nM–100 μM] or vehicle control for 72 h. Cell viability was measured using an MTT assay and normalised to vehicle control. N = 3 biological repeats each performed in triplicate. Error bars represent SEM. When error bars cannot be seen they are smaller than the symbols. ( F ) The graph shows the relative EC50 values for each cell type calculated using GraphPad Prism. Error bars represent SEM (* = p < 0.05 unpaired t -test). The inserted Western blot shows PRH levels in the CCA cell lines prior to treatment and with β-actin as a loading control.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_1616/pmc12651616/pmc12651616__cancers-17-03721-g001.jpg)
![SARS-CoV-2 virus infection in human airway epithelial cells in air:liquid interface, Vero E6, and endothelial cells. Human airway epithelial cells grown in an air:liquid interface (MucilAir) were infected with SARS-CoV-2 live virus (MOI = 0.1). Infectious virus released to the apical side of the epithelium was determined over time (6, 24, 48, and 72 h post-infection) ( A ). In separate studies, the levels of SARS-CoV-2 nucleocapsid or spike protein in Vero E6 and endothelial cells (treated with media only [untreated] or IL-1β [10 ng/mL; 3 h]) at 24 ( B ) and 72 ( C ) h post-infection with SARS-CoV-2 (MOI = 0.1) were determined using fluorescent imaging. Mock controls (media only) experiments were run simultaneously using each endothelial cell line. Data are shown as n = 3 (pooled donors) for Mucilair cells ( A ) and n = 3 (separate donors) for human aortic (AoEC), lung microvascular (HMVEC), and blood outgrowth endothelial cells (BOECs). Data are shown as mean ± SEM for (A) and representative images shown for (B) and (C) (scale bar = 25 µm).](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_4323/pmc12724323/pmc12724323__jvi.01205-25.f003.jpg)
