Journal: International Journal of Molecular Sciences

Article Title: Terconazole, an Azole Antifungal Drug, Increases Cytotoxicity in Antimitotic Drug-Treated Resistant Cancer Cells with Substrate-Specific P-gp Inhibitory Activity

doi: 10.3390/ijms232213809

Figure Lengend Snippet: Low treatment with TCZ, itraconazole, and posaconazole exert higher cytotoxic effects in P-gp-overexpressing cancer cells than that of other azole antifungal drugs. ( A ) KBV20C were treated with 5 nM VIC, the indicated μM concentrations of itraconazole (ITZ), TCZ, sertaconazole (STZ), sulconazole (SCZ), BTZ, econazole (ECZ), or posaconazole (PSZ) alone, or in combination with 5 nM VIC, or 0.1% DMSO (CON). After one day, all cells were observed using an inverted microscope at ×100 magnification. ( B , C ) KBV20C were treated with 5 nM VIC, 5 nM VIC with 2.5 μM itraconazole (VIC + ITZ), 5 nM VIC with 2.5 μM BTZ (VIC + BTZ), 5 nM VIC with 2.5 μM econazole (VIC + ECZ), 5 nM VIC with 2.5 μM sertaconazole (VIC + STZ), 5 nM VIC with 2.5 μM sulconazole (VIC + SCZ), 5 nM VIC with 2.5 μM ketoconazole (VIC + KTZ), 5 nM VIC with 2.5 μM oxiconazole (VIC + OXZ), or 0.1% DMSO (CON). After 24 h, annexin V analyses were performed as described in Materials and Methods. ( D , E ) KBV20C were treated with 5 nM VIC, 5 nM VIC with 2.5 μM itraconazole (VIC + ITZ), 5 nM VIC with 2.5 μM miconazole (VIC + MCZ), 5 nM VIC with 2.5 μM voriconazole (VIC + VCZ), 5 nM VIC with 2.5 μM fluconazole (VIC + FCZ), or 0.1% DMSO (CON). After one day, all cells were observed using an inverted microscope at ×100 magnification.

Article Snippet: TCZ, BTZ nitrate, itraconazole, posaconazole, econazole nitrate, ketoconazole, oxiconazole nitrate, sulconazole, sertaconazole, miconazole, voriconazole, fluconazole, aripiprazole, reserpine, and rifabutin were purchased from Selleckchem (Houston, TX, USA).

Techniques: Inverted Microscopy

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Sulconazole Induces PANoptosis by Triggering Oxidative Stress and Inhibiting Glycolysis to Increase Radiosensitivity in Esophageal Cancer

doi: 10.1016/j.mcpro.2023.100551

Figure Lengend Snippet: Sulconazole inhibits the viability of various cancer cell lines. A , molecular structure of sulconazole. B – D , viability of esophageal cancer cell lines (KYSE30 and KYSE150) and an esophageal epithelial cell line (SHEE) ( B ), liver cancer cell lines (HepG2 and Huh7) and a normal liver cell line (Chang liver) ( C ), gastric cancer cell lines (SGC7901 and HGC27), lung cancer cell line (A549), and breast cancer cell lines (MDA-MB-543 and MCF7) (D) were assessed after treatment with sulconazole at concentrations as indicated for 24 h. The data are representative of three independent experiments and presented as the mean ± SD.

Article Snippet: Sulconazole nitrate (HY-B1460A), Z-VAD-FMK (z-VAD) (HY-16658B), ferrostatin-1 (HY-100579), deferoxamine mesylate (HY-B0988), and necrostatin-1 (Nec) (HY-15760) were purchased from MedChemExpress and dissolved in dimethyl sulfoxide (DMSO) for storage, and the final concentration of DMSO used in all experiments did not exceed 0.1%.

Techniques:

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Sulconazole Induces PANoptosis by Triggering Oxidative Stress and Inhibiting Glycolysis to Increase Radiosensitivity in Esophageal Cancer

doi: 10.1016/j.mcpro.2023.100551

Figure Lengend Snippet: Sulconazole inhibits the proliferation and migration of esophageal cancer cell lines. A , cell clonogenic assay was used to verify the inhibition of proliferation of KYSE30 ( left panel ) and KYSE150 ( right panel ) cells after sulconazole treatment for 24 h. B , transwell assay was used to verify the inhibitory effect of sulconazole on migration of KYSE30 ( left panel ) and KYSE150 ( right panel) cells after sulconazole treatment for 24 h. The number of cells was counted in random fields. C and D , migration of KYSE30 and KYSE150 cells was detected by wound healing assay after sulconazole treatment for 24 h. Diagrams ( left panel ) were used for quantitative analyses of migration distance. All data are representative of three independent experiments. p -values were calculated by unpaired two-sided Student’s t tests. ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Sulconazole nitrate (HY-B1460A), Z-VAD-FMK (z-VAD) (HY-16658B), ferrostatin-1 (HY-100579), deferoxamine mesylate (HY-B0988), and necrostatin-1 (Nec) (HY-15760) were purchased from MedChemExpress and dissolved in dimethyl sulfoxide (DMSO) for storage, and the final concentration of DMSO used in all experiments did not exceed 0.1%.

Techniques: Migration, Clonogenic Assay, Inhibition, Transwell Assay, Wound Healing Assay

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Sulconazole Induces PANoptosis by Triggering Oxidative Stress and Inhibiting Glycolysis to Increase Radiosensitivity in Esophageal Cancer

doi: 10.1016/j.mcpro.2023.100551

Figure Lengend Snippet: Dysregulated proteins and pathways in esophageal cancer cells treated with sulconazole were analyzed by transcriptomics and proteomics. A , flowchart of the experimental process of transcriptomic sequencing and proteomic sequencing. B , Venn diagram of 1.5-fold change upregulated and downregulated genes from transcriptomic sequencing. C , HALLMARK enrichment analysis of dysregulated genes. D , KEGG enrichment analysis of dysregulated genes. E , volcano plot of 1.3-fold change upregulated and downregulated proteins from proteomic sequencing. F , KEGG enrichment analysis of dysregulated proteins. G , HALLMARK enrichment analysis of dysregulated proteins.

Article Snippet: Sulconazole nitrate (HY-B1460A), Z-VAD-FMK (z-VAD) (HY-16658B), ferrostatin-1 (HY-100579), deferoxamine mesylate (HY-B0988), and necrostatin-1 (Nec) (HY-15760) were purchased from MedChemExpress and dissolved in dimethyl sulfoxide (DMSO) for storage, and the final concentration of DMSO used in all experiments did not exceed 0.1%.

Techniques: Sequencing

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Sulconazole Induces PANoptosis by Triggering Oxidative Stress and Inhibiting Glycolysis to Increase Radiosensitivity in Esophageal Cancer

doi: 10.1016/j.mcpro.2023.100551

Figure Lengend Snippet: Sulconazole induces PANoptosis of esophageal cancer cells. A , Western blot analyses for the expression of BCL2, BAX, cleaved BAX, caspase3, cleaved caspase3, PARP, cleaved PARP, GSDME, GSDME-N, GSDMD, GSDMD-N, p-MLKL, MLKL, RIPK1, cleaved RIPK1, and β-Actin after sulconazole treatment for 12 h. B – D , flow cytometry ( B ) and quantification analysis ( C and D ) with Annexin V/PI staining evaluating the percentages of live cells in Q4 (Annexin V - /PI - ), early apoptotic cells in Q3 (Annexin V + /PI - ), and late apoptotic cells and necrotic cells in Q2 (Annexin V + /PI + ) among KYSE30 and KYSE150 treated with DMSO or sulconazole for 12 h. E , cell death in KYSE30 and KYSE150 cells treated with sulconazole for 12 h were detected by flow cytometry, and the PI-positive cells were calculated and shown in the diagrams. F , LDH release of KYSE30 and KYSE150 cells after sulconazole treatment for 12 h. G , heat map of ferroptosis-related genes that were differentially expressed in KYSE30 and KYSE150 cells with or without sulconazole treatment. H and I , the mRNA expression of ferroptosis-related genes by quantitative RT-PCR after sulconazole treatment for 24 h J – M , measurement of lipid peroxidation after sulconazole Ferr-1 (20 μM) and DFOM (100 μM) treatment for 12 h. Bar graph showing relative levels of lipid peroxidation by C11-BODIPY staining in KYSE30 and KYSE150 cells ( L and M ). N , representative images of cell death in KYSE30 and KYSE150 cells after sulconazole treatment for 12 h. O , diagrams were used for quantitative analyses of green dead cells (SYTOX Green-positive) in ( N ). The data in ( B – O ) are representative of three independent experiments and presented as the mean ± SD. p -values were calculated by two-way ANOVA. ∗∗ p < 0.01, ∗∗∗ p < 0.001. DFOM, deferoxamine mesylate; Ferr-1, ferrostatin-1; LDH, lactate dehydrogenase; MLKL, mixed lineage kinase-like domain; PI, propidium iodide.

Article Snippet: Sulconazole nitrate (HY-B1460A), Z-VAD-FMK (z-VAD) (HY-16658B), ferrostatin-1 (HY-100579), deferoxamine mesylate (HY-B0988), and necrostatin-1 (Nec) (HY-15760) were purchased from MedChemExpress and dissolved in dimethyl sulfoxide (DMSO) for storage, and the final concentration of DMSO used in all experiments did not exceed 0.1%.

Techniques: Western Blot, Expressing, Flow Cytometry, Staining, Quantitative RT-PCR

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Sulconazole Induces PANoptosis by Triggering Oxidative Stress and Inhibiting Glycolysis to Increase Radiosensitivity in Esophageal Cancer

doi: 10.1016/j.mcpro.2023.100551

Figure Lengend Snippet: Sulconazole triggers mitochondrial oxidative stress and inhibits glycolysis. A , transmission electron microscopy (TEM) images of KYSE30 cells subjected to the indicated treatments for 24 h. White arrows indicate mitochondria. Scale bars represent left, 1 μm; right, 500 nm. B ; PI, propidium iodide D , mitochondrial membrane potential analysis ( B ), ROS level analysis ( C ), and glucose uptake analysis ( D ) in KYSE30 and KYSE150 cells after sulconazole treatment for 24 h. E , heat map of 19 glycolysis-related enzymes that were differentially expressed in KYSE30 and KYSE150 cells with or without sulconazole treatment. F and G , the mRNA expression of glycolysis pathway enzymes by quantitative RT-PCR in KYSE30 ( F ) and KYSE150 ( G ) cells. H , Western blot analyses of glycolysis-related enzymes after sulconazole treatment for 24 h in KYSE30 ( left panel) and KYSE150 ( right panel ) cells. Quantification of the blots is shown below. I , Western blot analyses for the expression of p-AKT, AKT, p-MEK, MEK, p-ERK, ERK, p-STAT3, and STAT3 after sulconazole treatment for 24h in KYSE30 ( left panel ) and KYSE150 ( right panel) cells. The data in ( B – D , F and G ) are representative of three independent experiments and presented as the mean ± SD. p -values were calculated by two-way ANOVA. ∗∗ p < 0.01, ∗∗∗ p < 0.001. ROS, reactive oxygen species.

Article Snippet: Sulconazole nitrate (HY-B1460A), Z-VAD-FMK (z-VAD) (HY-16658B), ferrostatin-1 (HY-100579), deferoxamine mesylate (HY-B0988), and necrostatin-1 (Nec) (HY-15760) were purchased from MedChemExpress and dissolved in dimethyl sulfoxide (DMSO) for storage, and the final concentration of DMSO used in all experiments did not exceed 0.1%.

Techniques: Transmission Assay, Electron Microscopy, Membrane, Expressing, Quantitative RT-PCR, Western Blot

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Sulconazole Induces PANoptosis by Triggering Oxidative Stress and Inhibiting Glycolysis to Increase Radiosensitivity in Esophageal Cancer

doi: 10.1016/j.mcpro.2023.100551

Figure Lengend Snippet: Sulconazole increases radiosensitivity of esophageal cancer cells. A , flowchart of the clonogenic assay under sulconazole/IR chemoradiotherapy treatment. B , typical diagrams of radiation survival of KYSE30, KYSE150, and TE3 cells. C , D and E , statistical histogram of numbers of colonies formed. F , KYSE30 was treated with DMSO or sulconazole (20 μM) for 24 h, and γH2AX was detected at different time points after 4 Gy irradiation. G , ROS levels were detected after treatment with DMSO (control), sulconazole (20 μM), 4 Gy, or sulconazole (20 μM) combined with 4 Gy for 24 h in KYSE30 and KYSE150 cells. The data in ( C – E , G and H ) are representative of three independent experiments and presented as the mean ± SD. p -values were calculated by two-way ANOVA ( C – E ) and unpaired two-sided Student’s t test ( G and H ). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. IR, ionizing radiation; ROS, reactive oxygen species.

Article Snippet: Sulconazole nitrate (HY-B1460A), Z-VAD-FMK (z-VAD) (HY-16658B), ferrostatin-1 (HY-100579), deferoxamine mesylate (HY-B0988), and necrostatin-1 (Nec) (HY-15760) were purchased from MedChemExpress and dissolved in dimethyl sulfoxide (DMSO) for storage, and the final concentration of DMSO used in all experiments did not exceed 0.1%.

Techniques: Clonogenic Assay, Irradiation, Control

Journal: bioRxiv

Article Title: Deconvolution of polygenic risk score in single cells unravels cellular and molecular heterogeneity of complex human diseases

doi: 10.1101/2024.05.14.594252

Figure Lengend Snippet: a , Motif enrichment within HCM-CREs identified in two HCM-relevant cell types including cardiomyocyte and pericyte. Motif enrichment was measured by AUC. Row-wise standardization was performed. Only significant enrichment (adjusted P < 0.1, Bonferroni correction) is colored. CDM, cardiomyocyte; Peri, pericyte; AUC, the area under the receiver operating characteristic (ROC) curve. b , Bar plot of gene ontology (GO) enrichment for cardiomyocyte HCM risk genes. Significant GO terms (adjusted P < 0.1, BH correction) with odds ratio greater than five are shown. c , The network module M16 enriched with pericyte HCM risk genes. P -value by hypergeometric test. Edges between module genes are shown. d , Lollipop chart of GO enrichment (biological process) for M16 genes. Significant GO terms (adjusted P < 0.1, BH correction) are shown. e , Schematic of iPSC RNA-seq experiments. iPSC, induced pluripotent stem cell; Myk, Mavacamten; Omec, Omecamtiv mecarbil. f , Expression fold change comparison between HCM risk genes and the background transcriptome in cardiomyocytes across different conditions. The box plot center line, limits, and whiskers represent the median, quartiles, and 1.5x interquartile range (IQR), respectively. P -value by two-sided t -test. NS, not significant; stat, statistics. g , Expression fold change comparison between HCM risk genes and the background transcriptome in HCM-relevant cell types based on an HCM snRNA-seq study. P -value by two-sided t -test. h , Illustration of the genetic regulation of rs886125 in cardiomyocytes. Bar plot: “*” indicates scVINet-heart score percentage greater than 85%. Gene plot: differentially expressed target gene was mapped (in red). Bkg, background; Coaccess, coaccessibility. i , The uniform manifold approximation and projection (UMAP) plot of the left ventricle snRNA-seq dataset showing the expression of ZNF382 in individual cells. Expression was estimated by normalized gene count. Cardiomyocytes are highlighted in the dashed closed curve.

Article Snippet: Cells were treated with 250nM MYK-461 (Cayman Chemical, 19216-5mg), 400nM or 1uM omecamtiv mecarbil (Selleckchem via Fisher, NC1069600), or DMSO.

Techniques: RNA Sequencing Assay, Expressing, Comparison

Journal: bioRxiv

Article Title: Deconvolution of polygenic risk score in single cells unravels cellular and molecular heterogeneity of complex human diseases

doi: 10.1101/2024.05.14.594252

Figure Lengend Snippet: a , Lollipop chart of gene ontology (GO) enrichment (molecular function) for M16 genes. Significant GO terms (adjusted P < 0.1 by Benjamini-Hochberg (BH) correction) are shown. b , Expression fold change comparison between HCM pericyte genes and background transcriptome based on the HCM iPSC RNA-seq data. P -value by two-sided t -test. The box plot center line, limits, and whiskers represent the median, quartiles, and 1.5x interquartile range (IQR), respectively. CDM, cardiomyocyte; Myk, Mavacamten; Omec, Omecamtiv mecarbil; iPSC, induced pluripotent stem cell; NS, not significant. c , Expression analysis of HCM cardiomyocyte and pericyte genes based on the pericyte and cardiomyocyte transcriptome data, respectively. Peri, pericyte. d , The receiver operating characteristic curve (ROC; top) and the precision-recall curve (PRC; bottom) showing scVINet-heart test performance across various cell types in the human left ventricle. The red line and gray area represent the mean and 95% CI, respectively. The dashed gray line indicates the random prediction. AUC, the area under the curve; CI, confidence interval. e , Comparison of scVINet-heart prediction scores between eQTLs and other variants across different tissues and cell types. Comparison was performed using two-sided t -test. ×, adjusted P > 0.1 by BH correction. Rows were standardized. Mesothl, mesothelial cell; Adipo, adipose cell; CDM, cardiomyocyte; Fibro, fibroblast; LEC, lymphatic endothelial cell; Peri, pericyte; Schw, Schwann cell; SmMus, smooth muscle cell; VEC, vascular endothelial cell. f , Enrichment of transcription factor binding site (TFBS) disrupting variants within scVINet-heart-prioritized variants (various thresholds applied). Enrichment was quantified by t -statistics. ×, adjusted P > 0.1 by BH correction. g , Enrichment of scVINet-heart-prioritized HCM-associated variants within HCM-CREs. OR, odds ratio. OR and CI by two-sided Fisher’s exact test. OR is annotated by the solid line and 95% CI is represented by the shaded area. h-i , Summary statistics of prioritized HCM risk variants in cardiomyocytes ( h ) and pericytes ( i ) using different annotations.

Article Snippet: Cells were treated with 250nM MYK-461 (Cayman Chemical, 19216-5mg), 400nM or 1uM omecamtiv mecarbil (Selleckchem via Fisher, NC1069600), or DMSO.

Techniques: Expressing, Comparison, RNA Sequencing Assay, Binding Assay