Journal: iScience

Article Title: AURKB and PI3K/AKT/mTOR pathways converge to regulate TERT expression

doi: 10.1016/j.isci.2025.113194

Figure Lengend Snippet: Correlation and impact of AURKB expression on TERT levels in thyroid cancer subtypes (A) Thyroid cancer cells transfected with AURKB-specific shRNA plasmids were harvested following 4-day puromycin selection for assessment of AURKB levels by western blotting and TERT by RT-qPCR. Plotted values are the mean ± SD of three independent experiments, each performed in triplicate, expressed as fold-change in normalized expression relative to untransfected control. (B–F) Correlation between TERT and AURKB expression determined by RT-qPCR in TERTp mutated thyroid tumor cell lines: TPC1 (PTC), SW1736, 8505C, and C643 (ATC), alongside the non-malignant NThy-ori-3.1 cell line. Correlation between TERT and AURKB expression in (C) 220 PTC samples from the TCGA study, and (D) 37 advanced thyroid cancers from the Landa et al. study, using z-scores calculated from available transcriptomic data. Quantitative ChIP of AURKB, EZH2 and H3S10ph, H3K4Me3 and H3K27Me3 epigenetic marks at specific regions spanning the TERT transcriptional start site. Chromatin was prepared from (E) mutant C643 or TPC1, and (F) wild-type CAL62 and MDAT-T41 cells treated for 24 h with either 180 nM Hesperadin or DMSO. Plotted values are the mean average ± SD of three independent experiments. For all experiments, asterisks (∗) denote significant changes, determined at p < 0.05, using an unpaired two-tailed t -test with the assumption of equal variances.

Article Snippet: Human CAL62 anaplastic thyroid carcinoma cell line , CLS Cell Lines Service GmbH, Eppelheim, Germany , Cat#: 305114 RRID: CVCL_1112.

Techniques: Expressing, Transfection, shRNA, Selection, Western Blot, Quantitative RT-PCR, Control, Mutagenesis, Two Tailed Test

Journal: iScience

Article Title: AURKB and PI3K/AKT/mTOR pathways converge to regulate TERT expression

doi: 10.1016/j.isci.2025.113194

Figure Lengend Snippet: Impact of AURKB inhibition on TERT expression and telomerase activity (A–C) Dose response of TERT expression measured by RT-qPCR across thyroid cancer cell-lines treated with 0.1–10 μM Hesperadin. Plotted values are the mean ± SD of three independent experiments, each performed in triplicate, expressed as fold-change in normalized expression relative to DMSO control. Time-course analysis showing modulation of (B) TERT expression 24–72 h measured by RT-qPCR and (C) telomerase activity relative to MCF7 control 72–120 h measured by qTRAP, following treatment with IC50 dose of Hesperadin across various cancer cell lines. (D–F), Cell-cycle independent effect of Hesperadin on TERT expression was assessed by RT-qPCR at 0, 3, and 6 h post IC50 Hesperadin dose in SW1736 and CAL62 cells, which were arrested at the G2/M phase by Nocodazole. Flow cytometry was utilized to evaluate the cell cycle distribution of cells exposed to a 24 h Nocodazole vehicle, 24 h Nocodazole with a 6 h Hesperadin vehicle, or Nocodazole plus 6 h Hesperadin treatment. Data illustrates average cell percentages in G1, S, and G2/M phases (±SD). Values presented are mean ± SD from three independent experiments, each conducted in triplicate, expressed as fold changes in normalized expression/activity relative to 0 h baseline levels. For all experiments, asterisks (∗) denote significant changes, determined at p < 0.05, using an unpaired two-tailed t -test with the assumption of equal variances.

Article Snippet: Human CAL62 anaplastic thyroid carcinoma cell line , CLS Cell Lines Service GmbH, Eppelheim, Germany , Cat#: 305114 RRID: CVCL_1112.

Techniques: Inhibition, Expressing, Activity Assay, Quantitative RT-PCR, Control, Flow Cytometry, Two Tailed Test

Journal: iScience

Article Title: AURKB and PI3K/AKT/mTOR pathways converge to regulate TERT expression

doi: 10.1016/j.isci.2025.113194

Figure Lengend Snippet: mTORC1 inhibition leads to the dissociation of TRIM28-TRIM24 and AURKB from the mutant TERT promoter (A–F) Representative western blot images showing TRIM28 and TRIM24 expression across thyroid cell-lines. Co-immunoprecipitation of (B) endogenous TRIM24 in SW1736 and C643 cells; or (C) exogenous flag-tagged proteins overexpressed in SW1736 cells. Whole-cell lysates were incubated with anti-TRIM28 or IgG antibody for IP. (D, Left hand panel) Representative western blot images showing REST and expression across thyroid cell-lines. (D, Right hand panel) Co-immunoprecipitation of endogenous TRIM28 and AURKB by REST proteins in SW1736. Quantitative ChIP analysis was performed to measure TRIM28, TRIM24, AURKB, REST, EZH2 and H3S10ph levels at specific regions around the TERT transcriptional start site in (E) TPC1 and C643, and (F) CAL62 and MDAT-T41 cell following treatment with 2 nM Everolimus or DMSO for 6 h. The plotted values represent the mean ± SD derived from three independent experiments. In all experiments, asterisks (∗) indicate significant changes, determined at p < 0.05 using an unpaired two-tailed t -test with the assumption of equal variances.

Article Snippet: Human CAL62 anaplastic thyroid carcinoma cell line , CLS Cell Lines Service GmbH, Eppelheim, Germany , Cat#: 305114 RRID: CVCL_1112.

Techniques: Inhibition, Mutagenesis, Western Blot, Expressing, Immunoprecipitation, Incubation, Derivative Assay, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: CD47 and IFT57 Are Colinear Genes That Are Highly Coexpressed in Most Cancers and Exhibit Parallel Cancer-Specific Correlations with Survival

doi: 10.3390/ijms25168956

Figure Lengend Snippet: IFT57 and CD47 mRNA expression in normal and malignant thyroid epithelial cells and responses to transcriptional regulators. ( a – c ) MDA-T68 cells were treated with 10 μM JQ1 in DMSO or DMSO control for the indicated times. Expression of IFT57 and the long and short isoforms of CD47 mRNA were analyzed using real-time qPCR. ( d ) MDA-T68 cells were treated with 10 μM vemurafenib in DMSO or DMSO control for 24 h ( n = 3). Expression of IFT57 and the long and short isoforms of CD47 mRNA were analyzed using real-time qPCR. ( e ) Scatter plot showing linear regression analysis of CD47 and IFT57 mRNA levels in the 5 primary anaplastic thyroid carcinoma cell lines in CCLE.3.2. ( f – h ) in Nthy-ori 3-1 normal follicular thyroid epithelial cells, the follicular variant of papillary thyroid carcinoma cell line MDA-T68, and 8505C anaplastic thyroid carcinoma cell lines were analyzed for expression of IFT57 and long and short CD47 transcripts ( n = 3, (** = p < 0.01, *** = p < 0.001, **** = p < 0.0001, ns = not significant)) using default setting of GraphPad Prism software version 10.0.0..

Article Snippet: The 8505C undifferentiated anaplastic thyroid carcinoma cell line and Nthy-ori 3-1 normal human thyroid follicular epithelial cells were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Expressing, Control, Variant Assay, Software

Journal: International Journal of Molecular Sciences

Article Title: CD47 and IFT57 Are Colinear Genes That Are Highly Coexpressed in Most Cancers and Exhibit Parallel Cancer-Specific Correlations with Survival

doi: 10.3390/ijms25168956

Figure Lengend Snippet: Validation of CD47 and IFT57 mutants of the 8505C anaplastic thyroid carcinoma cell line. ( a ) Expression of IFT57 mRNA in WT 8505C and the sg2-B6 and sg1-F4 IFT57 knockdown mutants. ( b ) Expression of CD47 short mRNA in WT 8505C and the sg2-B6 and sg1-F4 IFT57 knockdown mutants. ( c ) Expression of IFT57 mRNA in WT 8505C and the CD47 knockdown mutant pool. ( d ) Flow cytometry analysis of cell surface CD47 expression in WT and mutant 8505C cell lines. ( e ) Western blot analysis of IFT57 and α-tubulin protein expression in WT and mutant 8505C cell lines. ( f ) Expression of PCNA mRNA in WT 8505C and the sg2-B6 and sg1-F4 IFT57 knockdown mutants. ( g ) Proliferation of WT 8505C cells and the sg2-B6 and sg1-F4 IFT57 knockdown mutants assessed using the MTS assay (mean ± SD, n = 6), (** = p ≤ 0.01, *** = p ≤ 0.001, **** = p ≤ 0.0001) using default setting of GraphPad Prism software version 10.0.0.

Article Snippet: The 8505C undifferentiated anaplastic thyroid carcinoma cell line and Nthy-ori 3-1 normal human thyroid follicular epithelial cells were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Expressing, Knockdown, Mutagenesis, Flow Cytometry, Western Blot, MTS Assay, Software

Journal: International Journal of Molecular Sciences

Article Title: CD47 and IFT57 Are Colinear Genes That Are Highly Coexpressed in Most Cancers and Exhibit Parallel Cancer-Specific Correlations with Survival

doi: 10.3390/ijms25168956

Figure Lengend Snippet: Analysis of differentially expressed genes identified by RNAseq in IFT57 and CD47 knockdown mutant 8505C cell lines and in TCGA papillary thyroid carcinomas. ( a ) Venn diagram showing overlap between differentially expressed genes with p < 0.05 in two IFT57 mutants versus WT (red) and the CD47 mutant pool versus WT 8505C cells (blue). ( b ) Of the 202 genes up-regulated or down-regulated >8-fold in 8505C IFT57 knockdown cells, 48 exhibited coexpression with IFT57 in TCGA papillary thyroid carcinomas with p < 10 −4 . Spearman’s correlations for coexpression of these genes with IFT57 and CD47 mRNA expression in TCGA papillary thyroid carcinomas are presented as a scatter plot. ( c – g ) Real-time qPCR validation of IFT57-dependent genes identified by RNSseq analysis. p -value ≤ 0.05 (*), ≤0.005 (**), ≤0.001 (***) was calculated using default settings of Bio-Rad CFX Maestro software 2.3. ( c ) Relative expression of RNF180 mRNA in parental 8505C cells (WT), the CD47 knockdown pool, and the indicated IFT57 knockdown clones. ( d ) Expression of nidogen-2 (NID2) mRNA in parental 8505C cells (WT), the CD47 knockdown pool, and the indicated IFT57 knockdown clones. ( e ) Expression of neuromedin-B (NMB) mRNA in parental 8505C cells (WT), the CD47 knockdown pool, and the indicated IFT57 knockdown clones. ( f ) Expression of CRACD mRNA in parental 8505C cells (WT), the CD47 knockdown pool, and the indicated IFT57 knockdown clones. ( g ) Expression of GAS6 mRNA in parental 8505C cells (WT), the CD47 knockdown pool, and the indicated IFT57 knockdown clones. ( h ) Intracellular flow cytometry of CRACD protein expression in parental 8505C cells (WT), the CD47 knockdown pool, and the indicated IFT57 knockdown clones.

Article Snippet: The 8505C undifferentiated anaplastic thyroid carcinoma cell line and Nthy-ori 3-1 normal human thyroid follicular epithelial cells were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Knockdown, Mutagenesis, Expressing, Software, Clone Assay, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: CD47 and IFT57 Are Colinear Genes That Are Highly Coexpressed in Most Cancers and Exhibit Parallel Cancer-Specific Correlations with Survival

doi: 10.3390/ijms25168956

Figure Lengend Snippet: Coexpression with IFT57 mRNA in papillary thyroid tumors for IFT57-dependent genes identified in 8505C cells. Genes with at least 8-fold changes in IFT57 knockdown cells were analyzed, and those with coexpression p -values < 10 −4 in papillary thyroid carcinomas are listed.

Article Snippet: The 8505C undifferentiated anaplastic thyroid carcinoma cell line and Nthy-ori 3-1 normal human thyroid follicular epithelial cells were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Knockdown, Expressing

Journal: Journal of Applied Genetics

Article Title: Occurrence of mammary gland tumours in male dogs and its weak association with development of testicular tumours: a review

doi: 10.1007/s13353-023-00818-z

Figure Lengend Snippet: Occurrence of mammary gland tumours (MGTs) in male dogs including benign neoplasms, carcinomas, and hyperplasia/dysplasia and accompanying testicular tumours (TTs) and/ or cryptorchidism with indicated immunohistological markers

Article Snippet: 66 , 5 × 4x3 , papillary , anaplastic mammary carcinoma , n.d , (Aslan et al. ) .

Techniques: Transformation Assay, In Situ