Journal: Molecular oncology

Article Title: The selenoenzyme type I iodothyronine deiodinase: a new tumor suppressor in ovarian cancer.

doi: 10.1002/1878-0261.13612

Figure Lengend Snippet: Fig. 1. Low selenoenzyme type I deiodinase (DIO1) levels in high-grade serous ovarian cancer (HGSOC) cell lines and xenografts. CHO-K1 (normal ovaries), FT282, FT109, ES-2, and Kuramochi cells were analyzed for the DIO1 by (A) flow cytometry and (B) Western blots. A representative analysis of three experimental repeats is presented. b-tubulin was used for protein loading. Average DIO1 protein expression SEM is presented. *P < 0.05 and **P < 0.05 by ANOVA for multiple comparisons. (C) Hematoxylin and eosin (H&E) and immunohistochemistry (IHC) staining for DIO1 and KI67 in formalin-fixed paraffin-embedded (FFPE) tumors from three mice. Scale bar, 20 lm. Matched isotype control is presented in Fig. S2A. 209 objective, Olympus microscopy (Olympus, Tokyo, Japan).

Article Snippet: Human HGSOC cells ES-2 (CVCL_3509) were purchased from the ATCC.

Techniques: Cytometry, Western Blot, Expressing, Immunohistochemistry, Control, Microscopy

Journal: Molecular oncology

Article Title: The selenoenzyme type I iodothyronine deiodinase: a new tumor suppressor in ovarian cancer.

doi: 10.1002/1878-0261.13612

Figure Lengend Snippet: Fig. 2. Selenoenzyme type I deiodinase (DIO1) correlates with overall survival in high-grade serous ovarian cancer (HGSOC) patients. DIO1 expression (high vs. low) was analyzed in (A) HGSOC patients or (B) patients treated with carboplatin/taxol, who underwent optimal (no residual tumor) or suboptimal debulking surgery. Data were retrieved from The Cancer Genome Atlas (TCGA) using the Kaplan–Meier Plotter tool (http://kmplot.com). HR, hazard ratio. (C) Boxplots of DIO1 expression in responders vs. non-responders, after therapy comple- tion using the ROC Plotter (https://rocplot.com/custom-data/index). *P value < 0.05 following Mann–Whitney test. (D) DIO1 immunohisto- chemistry (IHC) of formalin-fixed paraffin-embedded (FFPE) sections from human ovarian tumors (patients 1–10). Scale bar, 50 lm. Hematoxylin and eosin (H&E) and p53 staining were used to define tumor regions. p53 deleted cases (marked by an asterisk) underwent p16 staining. For each image, an enlarged inset of DIO1 expression is shown in the top right corner. Negative background stain was also confirmed using a matched isotype control antibody (Fig. S2B). 109 objective, Olympus microscopy (Olympus, Tokyo, Japan).

Article Snippet: Human HGSOC cells ES-2 (CVCL_3509) were purchased from the ATCC.

Techniques: Expressing, MANN-WHITNEY, Immunohistochemistry, Staining, Control, Microscopy

Journal: Molecular oncology

Article Title: The selenoenzyme type I iodothyronine deiodinase: a new tumor suppressor in ovarian cancer.

doi: 10.1002/1878-0261.13612

Figure Lengend Snippet: Fig. 3. Selenoenzyme type I deiodinase (DIO1) expression in normal ovary, fallopian tubes, and throughout high-grade serous ovarian cancer (HGSOC) evolution. Hematoxylin and eosin (H&E) and immunohistochemistry (IHC) analyses of p53, KI67, PAX8, and DIO1 in representative formalin-fixed paraffin-embedded (FFPE) tissues of (A) normal ovary and fallopian tubes and (B) serous tubal in-situ carcinoma (STIC), primary and colon metastasis from a representative HGSOC patient (patient #7). Scale bar, 50 lm. 109 objective, Olympus microscopy (Olympus, Tokyo, Japan).

Article Snippet: Human HGSOC cells ES-2 (CVCL_3509) were purchased from the ATCC.

Techniques: Expressing, Immunohistochemistry, In Situ, Microscopy

Journal: Molecular oncology

Article Title: The selenoenzyme type I iodothyronine deiodinase: a new tumor suppressor in ovarian cancer.

doi: 10.1002/1878-0261.13612

Figure Lengend Snippet: Fig. 4. Selenoenzyme type I deiodinase (DIO1) overexpression inhibits high-grade serous ovarian cancer (HGSOC) proliferation. ES-2 cells transiently transfected with empty PCDNA3 or DIO1-PCDNA3 expression vectors were analyzed after 3 days for (A) DIO1 protein levels in duplicates from control and DIO1 expression vectors are shown by Western blot (WB). b-tubulin was used for protein loading. Average DIO1 protein expression SEM is shown in the lower panel. (B) Light microscopy (X10 images, scale bar, 100 lm). Olympus microscopy images, using Cell^A software imaging. (C) Absolute cell counts, FC. (D) Annexin-PI histograms, FC. (E) Quantification of surviving cell population (Annexin/PI) and apoptotic cell population (Annexin+/PI+). Results (average SEM) were repeated twice, in triplicates. *P < 0.05 using Student’s unpaired t test.

Article Snippet: Human HGSOC cells ES-2 (CVCL_3509) were purchased from the ATCC.

Techniques: Over Expression, Transfection, Expressing, Control, Western Blot, Light Microscopy, Microscopy, Software, Imaging

Journal: Molecular oncology

Article Title: The selenoenzyme type I iodothyronine deiodinase: a new tumor suppressor in ovarian cancer.

doi: 10.1002/1878-0261.13612

Figure Lengend Snippet: Fig. 5. Selenoenzyme type I deiodinase (DIO1) silencing induces proliferation and alters protein expression in ovarian cancer. Analyses of DIO1 level in knockdown (KD) versus control ES-2 cells for (A) mRNA transcription (RQ-PCR). (B) Light microscopy of 1 9 105 control and DIO1-KD cells incubated in 24-well plates for 96 h (910 images, scale bar, 500 lm). Olympus microscopy images, using Cell^A software imaging. Isolated clones from DIO1-KD versus control cells were analyzed for (C) DIO1 protein level by Western blot. (D) Cell number, by flow cytometry. (E) pERK, p21, HK1, GAPDH, PKM2, PDHA1, ATP5A, and Trip11 levels by Western blot. b-tubulin was used for protein loading. (F) Proteomics analyses of differentially expressed proteins in DIO1-KD high-grade serous ovarian cancer (HGSOC). Interaction network, divided into three clusters, using STRING v. 10.5 (https://string-db.org), with default settings (minimum required interaction score: medium confidence 0.4). (G) Pie chart using PANTHER (http://pantherdb.org). Cellular and metabolic processes are shown. (H) Proteomaps using the bionic visualization tool (BionicVis). (I) Proteomics expression analyses for a collection of ATP5 synthase complex subunits in con- trol and DIO1-KD cells. Results (average SEM) are shown as % of control. Results were repeated twice in triplicates. *P < 0.05. **P < 0.005 by Student’s unpaired t test.

Article Snippet: Human HGSOC cells ES-2 (CVCL_3509) were purchased from the ATCC.

Techniques: Expressing, Knockdown, Control, Light Microscopy, Incubation, Microscopy, Software, Imaging, Isolation, Clone Assay, Western Blot, Cytometry

Journal: Molecular oncology

Article Title: The selenoenzyme type I iodothyronine deiodinase: a new tumor suppressor in ovarian cancer.

doi: 10.1002/1878-0261.13612

Figure Lengend Snippet: Fig. 6. Selenoenzyme type I deiodinase (DIO1) inhibition promotes ovarian cancer cell growth. ES-2 cells were seeded (1000 cells/96 wells), treated with PTU at increasing concentrations, and analyzed after 96 h for (A) cell density by light microscopy (910 images, scale bar, 100 lm). Olympus microscopy images, using Cell^A software imaging. (B) Cell proliferation (CyQUANT, ELISA). (C) Cell viability (PrestoBlue, ELISA). Next, normal ovary cells (CHO-K1), HGSOC cells (ES-2), and scrambled and DIO1-KD ES-2 cells were treated with 10 lM propylthiouracil (PTU) and analyzed by (D) Light microscopy (910 images, scale bar, 100 lm). Olympus microscopy images, using Cell^A software imaging. (E) Absolute cell counts (flow cytometry). (F) Cell viability (PrestoBlue, ELISA). Results (average SEM) were repeated twice in triplicates. *P < 0.05. **P < 0.005 using ANOVA.

Article Snippet: Human HGSOC cells ES-2 (CVCL_3509) were purchased from the ATCC.

Techniques: Inhibition, Light Microscopy, Microscopy, Software, Imaging, CyQUANT Assay, Enzyme-linked Immunosorbent Assay, Cytometry