type wt braf wt  (Addgene inc)

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Anaplastic thyroid cancer spheroids as preclinical models to test therapeutics

doi: 10.1186/s13046-024-03009-8

Figure Lengend Snippet: Comparison of gene expression between ATC cells from monolayer culture, spheroids culture, and parental tumor sample. ( A ) Heatmap showing gene expressions in ATC tumors and the matched monolayers and 3D spheroids. Genes with expressions that ( a ) have no significant difference between ATC tumors and matched 3D spheroids and ( b ) have a significant difference when comparing ATC tumors/matched 3D spheroids versus matched monolayers were selected. Red color represents higher expression while blue represents lower expression. ( B ) Heatmap showing upregulated gene expressions from comparison of 3D spheroids versus monolayers across 8505 C ATC cell line, C643 ATC cell line, and patient-derived ATC01 cells. ( C ) The RNA expression level of CDH1 and CDH2 in ATC01 cells. ( D ) The RNA expression level of CDH1 and CDH2 in 8505 C and C643 cells. ( E ) Pathway enrichment analysis for upregulated genes from ( A ). ( F ) Pathway enrichment analysis for upregulated genes from ( B )

Article Snippet: BRAF Wild Type (WT) cell line C643 was purchased from Cell Lines Service GmbH (Eppelheim, Germany) and THJ-16T derived from a patient with ATC was a kind gift from Dr. John A. Copland (Mayo Clinic, Jacksonville, FL).

Techniques: Comparison, Expressing, Derivative Assay, RNA Expression

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Anaplastic thyroid cancer spheroids as preclinical models to test therapeutics

doi: 10.1186/s13046-024-03009-8

Figure Lengend Snippet: Drug responses in anaplastic thyroid cancer (ATC) spheroid lines. ( A ) Workflow for evaluating cell viability in ATC spheroids using CellTiter-Glo 3D cell viability assay. ( B ) Representative images of ATC01 spheroids and monolayers in response to different drug treatments. ( C ) Drug-response analysis with dabrafenib, trametinib, and combination of both in ATC01 spheroids and monolayer cultured cells. ( D ) Representative images of 8505 C spheroids and monolayers in response to different drug treatments. ( E ) Drug-response analysis with dabrafenib, trametinib, and combination of both in 8505 C spheroids and monolayer cultured cells. ( F ) Representative images of C643 spheroids and monolayers in response to different drug treatments. ( G ) Drug-response analysis with dabrafenib, trametinib, and combination of both in C643 spheroids and monolayer cultured cells. Cell viability was measured by CellTiter-Glo assay after five days of drug treatment, and results were normalized to dimethyl sulfoxide-treated control cells. Each data point represents the mean ± SD of three technical duplicates. * P < 0.05 is the cell viability in the spheroid models compared to the cell viability in monolayer cultured cells, ns = nonsignificant. S = spheroids; M = monolayers; DAB = dabrafenib; TRA = trametinib; D + T = dabrafenib + trametinib, the combination ratio is 150:1, the dose-response curve of the combination group was plotted based on the doses of trametinib used in the treatment. The doses for trametinib in the combination group were 0.0085 µM, 0.017 µM, 0.033 µM, 0.067 µM, 0.133 µM. Based on the combination ratio (dabrafenib:trametinib = 150:1), the doses of dabrafenib in the combination group were 1.25 µM, 2.5 µM, 5 µM, 10 µM and 20 µM, respectively

Article Snippet: BRAF Wild Type (WT) cell line C643 was purchased from Cell Lines Service GmbH (Eppelheim, Germany) and THJ-16T derived from a patient with ATC was a kind gift from Dr. John A. Copland (Mayo Clinic, Jacksonville, FL).

Techniques: Viability Assay, Cell Culture, Glo Assay

Journal: Oncology Letters

Article Title: BRAF V600E -induced KRT19 expression in thyroid cancer promotes lymph node metastasis via EMT

doi: 10.3892/ol.2019.10360

Figure Lengend Snippet: The TCGA_THCA dataset was used for the following analysis. (A) Compared with the BRAF wild type group, the expression of KRT19 was increased in BRAF V600E group, however, not in BRAF wild type group. The (B) wild type BRAF or (C) V600E mutated BRAF overexpression plasmids were transfected into 8505C cells, and may be overexpressed significantly compared with their empty vectors, BRAF WT /BRAF V600E . (D) mRNA and (E) protein expression levels of the BRAF V600E mutation group exhibited higher expression levels of KRT19 in 8505C cells when compared with BRAF wild type group. BRAF, B-Raf Proto-Oncogene, Serine/Threonine Kinase; TCGA, The Cancer Genome Atlas Network; THCA, thyroid cancer; KRT19, keratin 19; WT, wild type.

Article Snippet: The wild type (WT) BRAF WT (pcDNA3.1, cat. no. 40775; Addgene, Inc.) with its empty vector were transfected to the cells followed by Lipofectamine 2000 instruction and mutant BRAF V600E (pBabe, cat. no. 15269; Addgene, Inc.) with its empty vector were transfected into retrovirus packaging phoenix cells (with an VSV.G envelope plasmid; cat. no. 14888; Addgene, Inc.), after 48 h culturing the virus supernatant was harvested and filtered.

Techniques: Expressing, Over Expression, Transfection, Mutagenesis