primary bronchial epithelial cells becs  (PromoCell)

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

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: Palbociclib decreases the viability of CCA cells but has less effect on primary biliary epithelial cells. CCA cell lines (CCLP ( A ), RBE ( B ), KKU-M055 ( C ), or primary BECs ( D )) were plated at 1 × 10 4 cells per well in a 96-well plate and treated with increasing concentrations of Palbociclib for 96 h. Cell viability was then measured using an MTT assay and normalised to the vehicle control. Three biological repeats (five for CCLP cells) each performed in triplicate. Error bars represent SEM. When error bars cannot be seen, they are smaller than the symbols. ( E ) The graph shows the relative EC50 values calculated using GraphPad Prism. Error bars represent SEM (** = p < 0.01 *** = p < 0.001 unpaired t -test).

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: MTT Assay, Control

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: Niclosamide and Pablociclib act synergistically to reduce CCA cell viability. ( A ) CCLP cells were plated at 1 × 10 4 cells per well in a 96-well plate before treatment with Niclosamide (NIC), Palbociclib (PAL), or both drugs in combination (COM) at the concentrations shown and for 72 h. Cell viability was then measured using an MTT assay. N = 3 biological experiments each performed in triplicate. Error bars represent SEM (* = p < 0.05, ** = p < 0.01, ns = not significant) two-tailed paired t -test. ( B ) From the data shown in ( A ) combination index (CI), values were calculated and plotted against the corresponding response (Fa) values in a CI plot. CI < 1 indicates synergism. ( C ) CCLP cells were treated with 0.5 μM Niclosamide (NIC), 1 μM Palbociclib (PAL), or both drugs in combination (COM) for 24 h, then proteins were extracted for Western blot. ( D ) CCLP cells and BECs were grown as spheroids for 5 days before treatment with 0.5 μM Niclosamide, 1 μM Palbociclib, or both drugs in combination for a further 72 h. Images were taken using a Nikon brightfield microscope, and spheroid size was calculated using ImageJ. N = 3 biological repeats. Error bars represent SEM (* = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001) two-tailed paired t -test. ( E ) Cell viability was measured in the spheroids from ( D ) using a PrestoBlue assay. Original western blots are presented in .

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: MTT Assay, Two Tailed Test, Western Blot, Microscopy, Prestoblue Assay

Journal: bioRxiv

Article Title: The RNA binding protein Insulin Growth factor 2 binding Protein 3 (IGF2BP3) modulates IL-13/IL-4 signalling in human bronchial epithelial cells and is dysregulated in type 2 disease

doi: 10.1101/2025.07.19.665669

Figure Lengend Snippet: IGF2BP3 is increased in type 2 disease. A. Left: normalized expression levels of IGF2BP3 mRNA in severe asthma (SA) vs healthy control (HC) primary BECs at total and polyribosome-bound levels; centre: IGF2BP3 normalized microarray expression data comparing airway epithelial cells from healthy non-atopic (HNA) and atopic asthma (AA) children; right IGF2BP3 normalized microarray expression data comparing nasal lavage samples from children during Picornavirus-induced exacerbation (EX) vs 7-14 days post-exacerbation (post-EX). B. Normalized count levels comparing type 2 high vs type 2 low nasal brushings for genes that showed IGF2BP3-dependent IL-13-modulation in our Frac-seq data . C: Heatmap showing the normalized expression values of the common mRNA signature between differentially expressed genes in the GALA II cohort and our Frac-seq IGF2BP3 and IL-13-dependent DEGs. D. Model for IGF2BP3 effects on type 2 responses. Decreasing IGF2BP3 levels leads to increased stability of IL13RA1 and IL4R mRNA and increased surface expression of IL4R and IL13Rα1, leading to increased STAT6 phosphorylation and increased IL-13 (and possibly IL-4) transcriptional responses, contributing to an overall increased type 2 response. In disease, IGF2BP3 is upregulated potentially to supress excessive T2 responses.

Article Snippet: Primary bronchial epithelial cells (BECs) were commercially sourced (PromoCell), grown and expanded in growth factor-supplemented medium (PromoCell) on collagen-coated plates.

Techniques: Expressing, Control, Microarray, Phospho-proteomics

Journal: bioRxiv

Article Title: The RNA binding protein Insulin Growth factor 2 binding Protein 3 (IGF2BP3) modulates IL-13/IL-4 signalling in human bronchial epithelial cells and is dysregulated in type 2 disease

doi: 10.1101/2025.07.19.665669

Figure Lengend Snippet: Decreasing IGF2BP3 levels increases IL-13/IL-4-driven transcriptional responses. RT-qPCR results of primary BECs transfected with siRNAs against IGF2BP3 (siIGF2BP3) or scrambled control (siControl) and stimulated with IL-13 or IL-4. A: RT-qPCR results for IGF2BP3 knock down and the major signalling mediators of IL-13 and IL-4. B: RT-qPCR results for chemokines. C: RT-qPCR results for pro-inflammatory IL6 and IL8 mRNAs. Data (n=3) depicted as mean ±SEM. One-tailed ratio t-tests. * P ≤ 0.05; ** P ≤ 0.01.

Article Snippet: Primary bronchial epithelial cells (BECs) were commercially sourced (PromoCell), grown and expanded in growth factor-supplemented medium (PromoCell) on collagen-coated plates.

Techniques: Quantitative RT-PCR, Transfection, Control, Knockdown, One-tailed Test

Journal: Clinical and Experimental Allergy

Article Title: Endothelin‐1 mediates Aspergillus fumigatus ‐induced airway inflammation and remodelling

doi: 10.1111/cea.13367

Figure Lengend Snippet: ET‐1 is up‐regulated in human bronchial epithelial cells exposed to Aspergillus fumigatus germinating spores. A, Confocal microscopy of live, germinating spores seeded onto BEC monolayers and stained with calcofluor‐white at 0, 12 and 18 h. Note the progressive emergence of hyphal extensions (Scale bar = 100 μm). B, In response to A fumigatus germinating spore exposure for 12 h, BECs increased gene expression of EDN1 (*** P < 0.001, n = 6) and pro‐inflammatory mediator, IL‐6 (* P ≤ 0.05, n = 6) as assessed by qPCR. Gene expression of other pro‐fibrogenic mediators, TGF‐β1 and POSTN , was unchanged whilst TGF‐β2 expression was significantly reduced (* P < 0.05, n = 6) relative to control. C, In response to A fumigatus germinating spores, BECs significantly increased the production of ET‐1 after 24 h (*** P < 0.001, n = 6)

Article Snippet: Human primary bronchial epithelial cells (BECs) were purchased from Promocell (Heidelberg, Germany) and Lonza (Basel, Switzerland).

Techniques: Confocal Microscopy, Staining, Expressing