human anaplastic thyroid cancer cell line 8505c  (Millipore)

Journal: bioRxiv

Article Title: CRISPR-Based Gene Dependency Screens reveal Mechanism of BRAF Inhibitor Resistance in Anaplastic Thyroid Cancer

doi: 10.1101/2025.06.26.661609

Figure Lengend Snippet: A. Human focused CRISPR/Cas9 KO library and CRISPR/dcas9 activation library containing number of guides were packed into lentiviral particles and transduced into Cas9+ or dCas9+ 8505c cells. The transduced cells were selected with puromycin. Newly generated mutant cell lines were cultured for 14 days with vehicle or dabrafenib (0.4 µM) for screening. Genomic DNA was extracted from the treated cells with vehicle or dabrafenib, sgRNA fragments were amplified by RT-PCR followed by next-generation sequencing. Genomic DNA from non-treated 8505c-transduced cell line was used as control. Sequencing data were analyzed using MAGeCKFlute. Genes were ranked by essentiality. B . Top 10 hits from the CRISPR/Cas9 KO at 14 days of dabrafenib treatment based on differential beta score . C. Top 50 “essential genes” from CRISPR/dCas9 activation screen. Red arrow pointing to HGF and blue arrow pointing to WWTR1. D . Venn Diagram showing the common essential genes from the CRISPR/Cas9 KO and activation screens’ top 50 hits. E. sgRNA targeting WWTR1 (TAZ) were depleted in dabrafenib treated cells and sgRNA activating WWTR1 were enriched in dabrafenib treated cells, suggesting that WWTR1 is an essential gene for survival during treatment. F . Cancer Dependency Map showing gene effect of WWTR1 using CHRONOS score in thyroid cancer cell lines . G . Gene expression Omnibus dataset (GSE33630) showed higher expression of WWTR1 in ATC compared to normal thyroid or PTC. H . WWTR1 (TAZ) essentiality in cancer cell lines based on BRAF mutation status using DepMap dataset. Essentiality score is defined by the effect of WWTR1 loss on cell viability. BRAFV600E-driven cancer cells are more sensitive to WWTR1 KO (TAZ KO) than non-BRAFV600E cells. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001

Article Snippet: The human anaplastic thyroid cancer cell line 8505c (Sigmal Millipore) was grown at 37°C with 5% CO 2 in Dulbecco’s Modified Eagle Medium (DMEM) (HyClone) supplemented with 10% Fetal Bovine Serum (Gemini Bio) and 1% Penicillin Streptomycin (Gibco).

Techniques: CRISPR, Activation Assay, Generated, Mutagenesis, Cell Culture, Amplification, Reverse Transcription Polymerase Chain Reaction, Next-Generation Sequencing, Control, Sequencing, Gene Expression, Expressing

Journal: bioRxiv

Article Title: CRISPR-Based Gene Dependency Screens reveal Mechanism of BRAF Inhibitor Resistance in Anaplastic Thyroid Cancer

doi: 10.1101/2025.06.26.661609

Figure Lengend Snippet: A. Depletion of TAZ using shRNA showed a mild effect on cell viability, However, it significantly reduced cell growth in the presence of dabrafenib B . Top and bottom panel, Colony formation assay showing that TAZ depletion sensitizes ATC cells to dabrafenib. C . TAZ-knock out markedly reduces colonies formation upon dabrafenib treatment . D. TAZ expression by western blot and RT PCR in shTAZ cells, and TAZ KO cells using CRISPR/Cas9 system and sgWWTR1-sg1 and sgWWTR1-sg2. E, F . TAZ-TEAD inhibitors, GNE-7933 and K-975 sensitized 8505c to dabrafenib. G . ATC patient-derived spheroids showed a significant reduction of cell viability in cell spheroids treated with the combination of GNE-7933 and dabrafenib compared to single agents. H . Immunoprecipitation assay showed a reduction of the binding of TAZ to TEAD with K-975 and GNE-7933. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: The human anaplastic thyroid cancer cell line 8505c (Sigmal Millipore) was grown at 37°C with 5% CO 2 in Dulbecco’s Modified Eagle Medium (DMEM) (HyClone) supplemented with 10% Fetal Bovine Serum (Gemini Bio) and 1% Penicillin Streptomycin (Gibco).

Techniques: shRNA, Colony Assay, Knock-Out, Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, CRISPR, Derivative Assay, Immunoprecipitation, Binding Assay

Journal: bioRxiv

Article Title: CRISPR-Based Gene Dependency Screens reveal Mechanism of BRAF Inhibitor Resistance in Anaplastic Thyroid Cancer

doi: 10.1101/2025.06.26.661609

Figure Lengend Snippet: A. Tumor growth curves of tumor xenografts in immunocompromised mice. 8505c-shCtrl or 8505c-shTAZ cells were implanted subcutaneously into the flanks. Treatment with vehicle or dabrafenib was initiated 2 weeks after tumor implantation. Dabrafenib was administered daily by gavage. Tumor diameters were measured weekly and used to calculate tumor volume. Data show mean±SEM ( n = 8 or 10). B . Tumor weight was recorded ex vivo. C . Animal weight recorded at the end of the experiment showing no weight loss due to dabrafenib treatment. D . Ex vivo images of the tumors harvested at the end point of the experiment E. Kaplan Meier survival curve after dabrafenib withdrawal. F . Growth curve over time of each tumor implanted for up to 3 weeks post drug withdrawal. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001

Article Snippet: The human anaplastic thyroid cancer cell line 8505c (Sigmal Millipore) was grown at 37°C with 5% CO 2 in Dulbecco’s Modified Eagle Medium (DMEM) (HyClone) supplemented with 10% Fetal Bovine Serum (Gemini Bio) and 1% Penicillin Streptomycin (Gibco).

Techniques: Tumor Implantation, Ex Vivo

Journal: bioRxiv

Article Title: CRISPR-Based Gene Dependency Screens reveal Mechanism of BRAF Inhibitor Resistance in Anaplastic Thyroid Cancer

doi: 10.1101/2025.06.26.661609

Figure Lengend Snippet: A. CBR-5884 sensitizes 8505c to dabrafenib and B . reduces PHGDH expression. C . GSK2606414 reduces ATF4 expression after 1 hour of treatment and sensitizes 8505c to dabrafenib. D, E . CBR-5884 and GSK2606414 sensitizes patient-derived spheroids (ATC-01) to dabrafenib after 10 days of treatment. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: The human anaplastic thyroid cancer cell line 8505c (Sigmal Millipore) was grown at 37°C with 5% CO 2 in Dulbecco’s Modified Eagle Medium (DMEM) (HyClone) supplemented with 10% Fetal Bovine Serum (Gemini Bio) and 1% Penicillin Streptomycin (Gibco).

Techniques: Expressing, Derivative Assay

Journal: Journal of the Endocrine Society

Article Title: Reevaluation of the Effect of Iodine on Thyroid Cell Survival and Function Using PCCL3 and Nthy-ori 3-1 Cells

doi: 10.1210/jendso/bvaa146

Figure Lengend Snippet: 131 Iodine ( 131 I) uptake in PCCL3, Nthy-ori 3-1, 8505C, and HepG2 cells, and thyrotropin receptor (TSHR) and sodium/iodine symporter (NIS) messenger RNA (mRNA) expression in PCCL3 and Nthy-ori 3-1 cells. A, Cells were cultured in the presence or absence of thyrotropin (TSH) and/or perchlorate and then incubated with 10 μCi/mL 131 I for 30 minutes. 131 I taken up into the cells was measured as described in “Materials and Methods.” Data are means ± SD (n = 3). B, Total RNA was extracted from PCCL3 and Nthy-ori 3-1 cells and subjected to RT-PCR for TSHR, NIS, and β-actin mRNAs as described in “Materials and Methods.” Data are means ± ranges (n = 2).

Article Snippet: Human anaplastic thyroid cancer cell line 8505C and human hepatocellular cancer cell line HepG2 were obtained from RIKEN Bioresource Center, cultured in RPMI 1640 with 10% FBS and antibiotics, and used as controls.

Techniques: Expressing, Cell Culture, Incubation, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of the Endocrine Society

Article Title: Reevaluation of the Effect of Iodine on Thyroid Cell Survival and Function Using PCCL3 and Nthy-ori 3-1 Cells

doi: 10.1210/jendso/bvaa146

Figure Lengend Snippet: Cytotoxicity of iodine in PCCL3, Nthy-ori 3-1, 8505C, and HepG2 cells. The cells, cultured in the presence or absence of perchlorate, were incubated with up to 5 × 10 –2 M iodine for 24 hours and cell viability was determined as described in “Materials and Methods.” Data are means ± SD (n = 3).

Article Snippet: Human anaplastic thyroid cancer cell line 8505C and human hepatocellular cancer cell line HepG2 were obtained from RIKEN Bioresource Center, cultured in RPMI 1640 with 10% FBS and antibiotics, and used as controls.

Techniques: Cell Culture, Incubation