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Supplementary Tables S1-S4 and Supplementary Figures S1-S12 from Digitalis-like Compounds Facilitate Non-Medullary Thyroid Cancer Redifferentiation through Intracellular Ca2+, FOS, and Autophagy-Dependent Pathways

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posted on 2023-04-03, 15:48 authored by Marika H. Tesselaar, Thomas Crezee, Herman G. Swarts, Danny Gerrits, Otto C. Boerman, Jan B. Koenderink, Hendrik G. Stunnenberg, Mihai G. Netea, Johannes W.A. Smit, Romana T. Netea-Maier, Theo S. Plantinga

Table S1. Overview of selected autophagy activating compounds, their structure, chemical and pharmacological classification and the supplier; Table S2. Significantly upregulated genes in all cell lines at all time points (fold change log2 {greater than or equal to} 1); Table S3. Significantly downregulated genes in all cell lines at all time points (fold change log2 {less than or equal to} -1); Table S4. Physicochemical properties of digitalis-like compounds predicted by ChemBioOf�ce software; Figure S1: Visualization of whole transcriptome data of BC-PAP treated with 50 µM proscillaridin A for 24, 48 or 72 hours, comprising principal component analysis and Venn diagrams of overlapping genes, either up- or downregulated, between 24, 48 and 72 hours time points; Figure S2: Visualization of whole transcriptome data of FTC133 treated with 50 µM digoxin for 24, 48 or 72 hours, comprising principal component analysis and Venn diagrams of overlapping genes, either up- or downregulated, between 24, 48 and 72 hours time points; Figure S3: Visualization of whole transcriptome data of FTC133 treated with 50 µM digoxigenin for 24, 48 or 72 hours, comprising principal component analysis and Venn diagrams of overlapping genes, either up- or downregulated, between 24, 48 and 72 hours time points; Figure S4: Visualization of whole transcriptome data of FTC133 treated with 50 µM proscillaridin A for 24, 48 or 72 hours, comprising principal component analysis and Venn diagrams of overlapping genes, either up- or downregulated, between 24, 48 and 72 hours time points; Figure S5: Visualization of whole transcriptome data of FTC133 treated with 50 µM strophantin K for 24, 48 or 72 hours, comprising principal component analysis and Venn diagrams of overlapping genes, either up- or downregulated, between 24, 48 and 72 hours time points; Figure S6: Visualization of whole transcriptome data of TPC-1 treated with lanatoside C for 24, 48 or 72 hours, comprising principal component analysis and Venn diagrams of overlapping genes, either up- or downregulated, between 24, 48 and 72 hours time points; Figure S7: STRING protein interaction networks of significantly up- or downregulated genes by digitalis-like compounds in all three cell lines and at all three time points; Figure S8: Bonferroni corrected P-value enrichment scores of significantly upregulated pathways by digitalis-like compounds combined for all three cell lines and for all three time points. P-values are generated by using the Gene Ontology (GO) platform category Biological Processes; Figure S9: Expression of JUN after treatment for 48 and 72 hours of BC-PAP, FTC133 and TPC-1 with the indicated digitalis-like compounds. Data are obtained from three independent experiments. Data are means {plus minus} SD. Square boxes represent statistical output of Spearman's rho tests for degree of correlation between expression of JUN with hNIS expression at 48 and 72 hours time points; Figure S10: Gene expression of thyroid transcription factors TTF1, TTF2 and PAX8 in BC-PAP, FTC133 or TPC-1 after treatment with the indicated digitalis-like compounds for 48 or 72 hours. Data are obtained from three independent experiments. Data are means {plus minus} SD; Figure S11: Heatmaps depicting gene expression of ABC transporters and organic anion transporters in untreated and digitalis-like compound treated BC-PAP, FTC133 and TPC-1 (N=6); Figure S12: Full unedited Western blot pictures, cropped pictures are depicted in Figure 1B.

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NWO

Dutch Cancer Society

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ARTICLE ABSTRACT

Up to 20%–30% of patients with metastatic non-medullary thyroid cancer have persistent or recurrent disease resulting from tumor dedifferentiation. Tumor redifferentiation to restore sensitivity to radioactive iodide (RAI) therapy is considered a promising strategy to overcome RAI resistance. Autophagy has emerged as an important mechanism in cancer dedifferentiation. Here, we demonstrate the therapeutic potential of autophagy activators for redifferentiation of thyroid cancer cell lines. Five autophagy-activating compounds, all known as digitalis-like compounds, restored hNIS expression and iodide uptake in thyroid cancer cell lines. Upregulation of hNIS was mediated by intracellular Ca2+ and FOS activation. Cell proliferation was inhibited by downregulating AKT1 and by induction of autophagy and p21-dependent cell-cycle arrest. Digitalis-like compounds, also designated as cardiac glycosides for their well-characterized beneficial effects in the treatment of heart disease, could therefore represent a promising repositioned treatment modality for patients with RAI-refractory thyroid carcinoma. Mol Cancer Ther; 16(1); 169–81. ©2016 AACR.