cis 4  (Gerstel GmbH)

Journal: Cancer Biology & Therapy

Article Title: Inhibition of WNT signaling sensitizes CRC cells to anti-tumor effects of retinoids: basis for new drug combinations in oncology

doi: 10.1080/15384047.2026.2732738

Figure Lengend Snippet: Evaluation of the drug combinations for anti-proliferative effects on different CRC cell lines. The response of drug combinations was further evaluated for antagonistic, additive, and synergistic responses on CRC cell proliferation. The combination effects were calculated as a ratio of the observed response divided by the predicted response based on the Bliss independence model. ⁵² Panel A shows that 9-cis-RA + Sulindac and Piroxicam, Liarozole + Piroxicam, and ATRA + Sulindac have additive effects in HT29 cells. Panel B shows that 9-cis-RA + Sulindac and Piroxicam, Liarozole + Piroxicam, and ATRA + Sulindac have synergistic effects in HCT116 cells. Panel C shows that Talarozole + Sulindac have synergistic effects in SW480 cells. The synergistic response is < 1, additive ~1, and antagonistic > 1.

Article Snippet: Drug sources are as follows: Sulindac, Piroxicam, and Talarozole (Sigma-Aldrich, St Louis, MO), 9-cis retinoic acid (Santa Cruz Biotechnology, Dallas, TX), Liarozole (R&D Chemical, Gonzales, LA), and all-trans retinoic acid (Stem Cell Technologies, Vancouver, BC).

Techniques:

Journal: Cancer Biology & Therapy

Article Title: Inhibition of WNT signaling sensitizes CRC cells to anti-tumor effects of retinoids: basis for new drug combinations in oncology

doi: 10.1080/15384047.2026.2732738

Figure Lengend Snippet: Drug combination effects on stem cells and neuroendocrine cells. The effects of retinoids and anti-WNT agents on ALDH + stem cells were evaluated based on the ALDEFLUOR assay. The results showed that treatment with the 9-cis RA + Sulindac drug combination substantially decreased the population of ALDH + cells in HCT116 cells (Panel B), but not in HT29 (Panel A) or SW480 (Panel C) cells. Notably, this decrease was seen in HCT116 cells at both time points (24 and 48 h). Neuroendocrine cells (NECs). The effects of retinoids and anti-WNT agents on NEC differentiation were evaluated using western blot analysis for expression of differentiation markers in HCT116 CRC cells (Panels D–F) and SW480 cells (Panels G–I). The results show that drug combinations increase NEC differentiation in HCT116 and SW480 CRC cells. GLP2R = glucagon-like protein two receptors, SSTR1 = somatostatin receptor 1, CHGA = chromogranin A, NSE = neuron-specific enolase. The experiments were done in triplicate, and the error bars represent ± the SEM. * p < 0.05.

Article Snippet: Drug sources are as follows: Sulindac, Piroxicam, and Talarozole (Sigma-Aldrich, St Louis, MO), 9-cis retinoic acid (Santa Cruz Biotechnology, Dallas, TX), Liarozole (R&D Chemical, Gonzales, LA), and all-trans retinoic acid (Stem Cell Technologies, Vancouver, BC).

Techniques: Western Blot, Analysis, Expressing

Journal: Cancer Biology & Therapy

Article Title: Inhibition of WNT signaling sensitizes CRC cells to anti-tumor effects of retinoids: basis for new drug combinations in oncology

doi: 10.1080/15384047.2026.2732738

Figure Lengend Snippet: The effect of drug combinations on changes in gene expression in CRC cells. We conducted NanoString Profiling of HCT116 and SW480 cells treated with 9-cis-RA and Liarozole in combination with Sulindac to study the effects of drug combinations on gene expression. The heat maps (Panels A & B) show that drug combination treatments involving an RA agent (9-cis RA or Liarozole) in combination with an anti-WNT agent (Sulindac) lead to dramatic differences in gene expression compared to treatment with single agents. Sulindac treatment leads to the downregulation of a substantial number of genes in HCT116 cells (Panel C) and SW480 cells (Panel D). This down-regulation is characterized by changes in the expression of various WNT target genes. Changes in the expression of genes with WNT-regulatory elements (WREs) in HCT116 (Panels A and C) and SW480 (Panels B and D) cells indicate that Sulindac targets WNT signaling. Genes with WNT-regulatory elements show up- or down-regulation of expression (>2X-fold) in response to Sulindac treatment in HCT116 (Panel C) and SW480 (Panel D) CRC cells. In HCT116 cells, decreased expression of the notable WNT target genes MET , MYC , AXIN2 , BMP4 , RNF43 , and TCF3 are shown (Panel E). While in SW480 cells, the expression of the WNT target genes AXIN2 , BMP4 , RNF43 , and CCND1 had decreased (Panel F).

Article Snippet: Drug sources are as follows: Sulindac, Piroxicam, and Talarozole (Sigma-Aldrich, St Louis, MO), 9-cis retinoic acid (Santa Cruz Biotechnology, Dallas, TX), Liarozole (R&D Chemical, Gonzales, LA), and all-trans retinoic acid (Stem Cell Technologies, Vancouver, BC).

Techniques: Gene Expression, Profiling, Expressing

Journal: Cancer Biology & Therapy

Article Title: Inhibition of WNT signaling sensitizes CRC cells to anti-tumor effects of retinoids: basis for new drug combinations in oncology

doi: 10.1080/15384047.2026.2732738

Figure Lengend Snippet: Analysis of Sulindac’s ability to sensitize CRC cells to the effect of retinoids based on gene expression. Further NanoString expression profiling was done on HCT116 and SW480 cells treated with 9-cis-RA or Liarozole in combination with Sulindac. The effect of RA agents on CRC cells . The Venn diagrams (Panel A) show that changes in the expression ( p < 0.05) of the same genes in both cell lines (HCT116 and SW480) occurred with 9-cis-RA treatment ( n = 38 genes) and Liarozole treatment ( n = 45 genes). The Venn diagrams (Panel B) also show that changes in the expression ( p < 0.05) of the same genes occur with both RA agents (9-cis-RA and Liarozole) in HCT116 ( n = 49 genes) and SW480 cells ( n = 65 genes). These results indicate that a common set of genes is targeted by both of the RA agents (9-cis-RA and Liarozole) in CRC cells. Indeed, 19 of the same genes changed in expression (7 upregulated and 12 down-regulated) with both RA agent treatments and in both CRC cell lines (Panel C). The effect of Sulindac on responses to RA agents . In both HCT116 (Panels D and E) and SW480 (Panels F and G) cells, one set of genes is down-regulated, and another set is up-regulated ( p < 0.05) by retinoid agents after Sulindac treatment. Additional analyses to illustrate the change in the expression of the specific genes are presented in waterfall plots (Figure S3) and in lists of the top 10 genes upregulated (Table S3) and downregulated (Table S4) by RA agents after Sulindac treatment. Interestingly, many of the same genes are upregulated or downregulated by Liarozole and 9-cis RA in both cell lines. Overall, these results indicate that Sulindac sensitizes HCT116 and SW480 CRC cells to the effect of retinoids on gene expression.

Article Snippet: Drug sources are as follows: Sulindac, Piroxicam, and Talarozole (Sigma-Aldrich, St Louis, MO), 9-cis retinoic acid (Santa Cruz Biotechnology, Dallas, TX), Liarozole (R&D Chemical, Gonzales, LA), and all-trans retinoic acid (Stem Cell Technologies, Vancouver, BC).

Techniques: Analysis, Gene Expression, Expressing, Profiling

Journal: Cancer Biology & Therapy

Article Title: Inhibition of WNT signaling sensitizes CRC cells to anti-tumor effects of retinoids: basis for new drug combinations in oncology

doi: 10.1080/15384047.2026.2732738

Figure Lengend Snippet: The two sets of retinoid-response genes (down-regulated vs. up-regulated) have different functional classifications. *

Article Snippet: Drug sources are as follows: Sulindac, Piroxicam, and Talarozole (Sigma-Aldrich, St Louis, MO), 9-cis retinoic acid (Santa Cruz Biotechnology, Dallas, TX), Liarozole (R&D Chemical, Gonzales, LA), and all-trans retinoic acid (Stem Cell Technologies, Vancouver, BC).

Techniques: Functional Assay, Transduction, Tissue, Cell Differentiation

Journal: Cancer Biology & Therapy

Article Title: Inhibition of WNT signaling sensitizes CRC cells to anti-tumor effects of retinoids: basis for new drug combinations in oncology

doi: 10.1080/15384047.2026.2732738

Figure Lengend Snippet: Measuring drug responses using organoid cultures of human CRC cells. The effect of therapeutic manipulation of WNT and RA signaling was evaluated in human patient-derived colon cancer organoids. Drug-response experiments were conducted using a CRC organoid model from the ATCC (HCM-CSHL-0257-C18). Treatment concentrations were chosen based on the effective IC50 doses as indicated: 9-cis-RA (90 µM), Sulindac (322.5, 430, 537.5 µM), and Liarozole (90 µM). Panels A & B show representative microscopy staining from cell viability analysis using CYTO 3D Live/Dead Assay Kit. Scale bars equal 50 µM. Each panel contains four rows of images. Brightfield shows the overall cell morphology under normal light. AO (acridine orange): green fluorescence indicating live cells. PI (propidium iodide): red fluorescence marks dead cells. Merged: A combination of AO and PI channels showing both live and dead cells simultaneously.Panels C and D give the quantification for cell death based on the PI (dead cells)/AO (live cells) ratio. The cells were treated with various concentrations of 9-cis-RA, Liarozole, Sulindac alone, or combinations of both compounds. The control represents untreated cells. Increasing ratios indicate higher proportions of dead cells than live cells, with combination treatments showing the most pronounced effects. This experiment was done in triplicate, and the error bars represent ± the SEM. Overall, treating CRC organoid cultures with anti-WNT & RA drug combinations causes a substantial increase in cell death compared to individual agents alone.

Article Snippet: Drug sources are as follows: Sulindac, Piroxicam, and Talarozole (Sigma-Aldrich, St Louis, MO), 9-cis retinoic acid (Santa Cruz Biotechnology, Dallas, TX), Liarozole (R&D Chemical, Gonzales, LA), and all-trans retinoic acid (Stem Cell Technologies, Vancouver, BC).

Techniques: Derivative Assay, Organoids, Microscopy, Staining, Analysis, Live Dead Assay, Brightfield, Morphology, Fluorescence, Control

Journal: Cancer Biology & Therapy

Article Title: Inhibition of WNT signaling sensitizes CRC cells to anti-tumor effects of retinoids: basis for new drug combinations in oncology

doi: 10.1080/15384047.2026.2732738

Figure Lengend Snippet: Drug combination responses in a human CRC PDX (CTG-2359) mouse model. Drug safety . Drug toxicity was initially evaluated for tolerability (Panel A), and during drug treatments for efficacy. Different colored lines indicate various treatment conditions, including control and combination treatments. Body weights remained mostly stable with no weight loss greater than 10%. This reveals that the drug treatments were reasonably well tolerated. Efficacy analysis . Panel B shows the time course results on tumor volume changes in the efficacy study over 28 d. The treatment groups included a placebo control and various Sulindac, Liarozole, and 9-cis RA combinations. The tumor volumes were measured in cubic millimeters and plotted against time (days). The error bars represent ± the SEM. All the treatment groups showed progressive tumor growth, with the combinations of Sulindac + Liarozole + 9-cis RA demonstrating the lowest tumor volume trajectory. The placebo group exhibited the highest tumor growth rate, reaching approximately 2000 mm 3 by day 25. Measurements were taken at 5-day intervals after treatment initiation at day 0. Panel C. The waterfall plot shows the percent decreased in tumor growth for all treatment groups. The corresponding values for anti-tumor activity are provided in Table 3 below.

Article Snippet: Drug sources are as follows: Sulindac, Piroxicam, and Talarozole (Sigma-Aldrich, St Louis, MO), 9-cis retinoic acid (Santa Cruz Biotechnology, Dallas, TX), Liarozole (R&D Chemical, Gonzales, LA), and all-trans retinoic acid (Stem Cell Technologies, Vancouver, BC).

Techniques: Toxicity, Control, Analysis, Activity Assay

Journal: Cancer Biology & Therapy

Article Title: Inhibition of WNT signaling sensitizes CRC cells to anti-tumor effects of retinoids: basis for new drug combinations in oncology

doi: 10.1080/15384047.2026.2732738

Figure Lengend Snippet: Anti-tumor activity for human CRC PDX (CTG-2359) mouse model.

Article Snippet: Drug sources are as follows: Sulindac, Piroxicam, and Talarozole (Sigma-Aldrich, St Louis, MO), 9-cis retinoic acid (Santa Cruz Biotechnology, Dallas, TX), Liarozole (R&D Chemical, Gonzales, LA), and all-trans retinoic acid (Stem Cell Technologies, Vancouver, BC).

Techniques: Activity Assay, Control