Journal: Cancer letters

Article Title: Synergistic efficacy of irinotecan and sunitinib combination in preclinical models of anaplastic thyroid cancer

doi: 10.1016/j.canlet.2017.09.032

Figure Lengend Snippet: Antiproliferative effects of sunitinib (SU) and SN-38 in vitro on 8305C (A and C, respectively), and FB3 (B and D, respectively) cell lines. The antiproliferative effects of the drugs were studied after 72 h of exposure. The data are presented as percentage of vehicle-treated cells. The concentrations of drug that reduced cell proliferation by 50% (IC50) vs controls were calculated by a nonlinear regression fit of the mean values of the data obtained in triplicate experiments (i.e. at least 9 wells for each concentration). Columns and bars, mean values ± S.E., respectively. *, P < 0.001 vs. control.

Article Snippet: The human ATC cell line 8305C ( BRAF V600E mutated) – established from undifferentiated thyroid carcinomas of a 67 year-old-female patient – was from DSMZ (Braunschweig, Germany, DSMZ no.: ACC 133) [ 18 ], whereas the human ATC cell line FB3 ( HRas Q61R mutated) was obtained from Prof. Fulvio Basolo of the University of Pisa, Pisa, Italy [ 6 ].

Techniques: In Vitro, Concentration Assay

Journal: Cancer letters

Article Title: Synergistic efficacy of irinotecan and sunitinib combination in preclinical models of anaplastic thyroid cancer

doi: 10.1016/j.canlet.2017.09.032

Figure Lengend Snippet: CI (Combination Index) and DRI (Dose Reduction Index) values for the drug combinations at 25%, 50%, and 75% levels of inhibition of 8305C and FB3 cell proliferation. SN-38, active metabolite of irinotecan; SU, sunitinib.

Article Snippet: The human ATC cell line 8305C ( BRAF V600E mutated) – established from undifferentiated thyroid carcinomas of a 67 year-old-female patient – was from DSMZ (Braunschweig, Germany, DSMZ no.: ACC 133) [ 18 ], whereas the human ATC cell line FB3 ( HRas Q61R mutated) was obtained from Prof. Fulvio Basolo of the University of Pisa, Pisa, Italy [ 6 ].

Techniques: Inhibition

Journal: Cancer letters

Article Title: Synergistic efficacy of irinotecan and sunitinib combination in preclinical models of anaplastic thyroid cancer

doi: 10.1016/j.canlet.2017.09.032

Figure Lengend Snippet: Accumulation of SN-38 (ng · mg−1 protein) in 8305C (A) and FB3 (B) cell lines after exposure to 1 μM SN-38 alone and in combination with sunitinib (SU). Columns and bars indicate the mean percentage values (±S.D.) vs. treated cells with SN-38 alone. ABCG2 gene expression (2−ΔΔCt) and ABCG2 (ng · mg−1 protein) protein levels in 8305C (C and E, respectively) and FB3 (D and F, respectively) cells exposed to sunitinib or with vehicle alone for 72 h. Data are expressed as percentage of vehicle-treated cells. Columns and bars, mean values ± S.D., respectively. *P < 0.05 vs. vehicle-treated controls. The quantitation of gene expression was performed using the ΔΔCt calculation, where Ct is the threshold cycle; the amount of target, normalized to the endogenous control, glyceraldehyde 3-phosphate dehydrogenase, and relative to the calibrator (vehicle treated control cells), is given as 2−ΔΔCt. The quantitation of protein levels was performed by ELISA. The optical density was determined using a Multiskan Spectrum microplate reader set to 450 nm. The results were expressed as nanograms of ABCG2 per milligram of total protein. All experiments were repeated, independently, three times.

Article Snippet: The human ATC cell line 8305C ( BRAF V600E mutated) – established from undifferentiated thyroid carcinomas of a 67 year-old-female patient – was from DSMZ (Braunschweig, Germany, DSMZ no.: ACC 133) [ 18 ], whereas the human ATC cell line FB3 ( HRas Q61R mutated) was obtained from Prof. Fulvio Basolo of the University of Pisa, Pisa, Italy [ 6 ].

Techniques: Expressing, Quantitation Assay, Enzyme-linked Immunosorbent Assay

Journal: Cancer letters

Article Title: Synergistic efficacy of irinotecan and sunitinib combination in preclinical models of anaplastic thyroid cancer

doi: 10.1016/j.canlet.2017.09.032

Figure Lengend Snippet: VEGF (A) and CSF-1 (B) gene expression (2−ΔΔCt) in 8305C and FB3 cells and (C) CSF-1 (ng · mg−1 protein) protein levels in FB3 cells exposed to sunitinib (SU) or with vehicle alone for 72 h. Columns and bars, mean values ± S.D., respectively. *P < 0.05 vs. vehicle-treated controls. Amplifications were normalized to glyceraldehyde 3-phosphate dehydrogenase, and the quantitation of gene expression was performed using the ΔΔCt calculation, where Ct is the threshold cycle; the amount of target, normalized to the endogenous control and relative to the calibrator (vehicle-treated control cells), is given as 2−ΔΔCt. The quantitation of protein levels was performed by ELISA. The optical density was determined using a Multiskan Spectrum microplate reader set to 450 nm. The results were expressed as nanograms of CSF-1 per milligram of total protein. All experiments were repeated, independently, three times.

Article Snippet: The human ATC cell line 8305C ( BRAF V600E mutated) – established from undifferentiated thyroid carcinomas of a 67 year-old-female patient – was from DSMZ (Braunschweig, Germany, DSMZ no.: ACC 133) [ 18 ], whereas the human ATC cell line FB3 ( HRas Q61R mutated) was obtained from Prof. Fulvio Basolo of the University of Pisa, Pisa, Italy [ 6 ].

Techniques: Expressing, Quantitation Assay, Enzyme-linked Immunosorbent Assay

Journal: Cancer letters

Article Title: Synergistic efficacy of irinotecan and sunitinib combination in preclinical models of anaplastic thyroid cancer

doi: 10.1016/j.canlet.2017.09.032

Figure Lengend Snippet: In vivo antitumor effects of the single drugs and three different combination schedules of sunitinib (SU) and irinotecan (CPT-11) on 8305C tumors xenotransplanted in mice (A); Immunohistochemistry quantification of CD31 (B) and Capase-3 positive cells (C) in 8305C tumor xenografts administered with vehicle, SU at 25 mg/kg every 3 days, CPT-11 100 mg/kg every week through i.p. injection, and their combinations. Symbols/columns and bars, mean values ± S.D., respectively.*P < 0.001 vs. vehicle-treated controls. #P < 0.001 vs. sunitinib-treated group.

Article Snippet: The human ATC cell line 8305C ( BRAF V600E mutated) – established from undifferentiated thyroid carcinomas of a 67 year-old-female patient – was from DSMZ (Braunschweig, Germany, DSMZ no.: ACC 133) [ 18 ], whereas the human ATC cell line FB3 ( HRas Q61R mutated) was obtained from Prof. Fulvio Basolo of the University of Pisa, Pisa, Italy [ 6 ].

Techniques: In Vivo, Immunohistochemistry, Injection

Journal: Asian Pacific Journal of Cancer Prevention

Article Title: Targeting the BRAF Signaling Pathway in CD133pos Cancer Stem Cells of Anaplastic Thyroid Carcinoma

doi: 10.31557/apjcp.2019.20.5.1353

Figure Lengend Snippet: Figure 1. Results of Flow Cytometry Method. Flow cytometry assay confirmed the purity and sorting of the CD133pos cells derived from ATC cell lines (C643, SW1736 and 8305C, respectively) after using MACS method (D-F) compared to before isolation (A-C).

Article Snippet: Anaplastic Thyroid Carcinoma cell lines and cell culture The ATC cell line (8305C) was bought from the National Cell Bank of Iran (Pasteur Institute of Iran, Tehran, Iran).

Techniques: Flow Cytometry, Derivative Assay, Isolation

Journal: Asian Pacific Journal of Cancer Prevention

Article Title: Targeting the BRAF Signaling Pathway in CD133pos Cancer Stem Cells of Anaplastic Thyroid Carcinoma

doi: 10.31557/apjcp.2019.20.5.1353

Figure Lengend Snippet: Figure 2. Evaluate the BRAF Gene Expression in Human Normal Thyrocytes, ATC Cell Lines and the CD133pos Cells Isolated from them. Data (Mean ± SEM, n=3) showed that despite this gene has not any level expression in human normal thyrocytes (A), has a huge expression level in the CD133pos cells isolated from C643(C, 1705.99 ± 55.55), SW1736 (E, 7265.85 ± 511.91), and 8305C (G, 6543.95 ± 136.15) cell lines. In C643, SW1736, and 8305C cell lines, expression levels of this gene were 509.87 ± 52.28 (B), 2461.43 ± 310.91 (D), and 1303.56 ± 148.15 (F), respectively. GAPDH housekeeping gene was used for mRNAs levels normalization. ⃰ P<0.05 by One-way ANOVA test.

Article Snippet: Anaplastic Thyroid Carcinoma cell lines and cell culture The ATC cell line (8305C) was bought from the National Cell Bank of Iran (Pasteur Institute of Iran, Tehran, Iran).

Techniques: Gene Expression, Isolation, Expressing

Journal: Asian Pacific Journal of Cancer Prevention

Article Title: Targeting the BRAF Signaling Pathway in CD133pos Cancer Stem Cells of Anaplastic Thyroid Carcinoma

doi: 10.31557/apjcp.2019.20.5.1353

Figure Lengend Snippet: Figure 3. Evaluate the NIS Gene Expression in Human Normal Thyrocytes, ATC Cell Lines and the CD133pos Cells Isolated from them. Data (Mean ± SEM, n=3) showed that NIS mRNA levels are very low in C643 (B, 0.29 ± 0.04), SW1736 (D, 0.29 ± 0.11), and 8305C (F, 0.35 ± 0.03) cell lines compared to human normal thyrocytes (A). Also it has a tiny level in CD133pos cells segregated from C643(C, 0.0003 ± 2.71), SW1736 (E, 0.0002 ± 0.71), and 8305C (G, 0.0008 ± 5.43) cell lines, respectively. GAPDH gene was used for mRNAs levels normalization. ⃰ P<0.05 by One-way ANOVA test.

Article Snippet: Anaplastic Thyroid Carcinoma cell lines and cell culture The ATC cell line (8305C) was bought from the National Cell Bank of Iran (Pasteur Institute of Iran, Tehran, Iran).

Techniques: Gene Expression, Isolation

Journal: Asian Pacific Journal of Cancer Prevention

Article Title: Targeting the BRAF Signaling Pathway in CD133pos Cancer Stem Cells of Anaplastic Thyroid Carcinoma

doi: 10.31557/apjcp.2019.20.5.1353

Figure Lengend Snippet: Figure 4. Results of BRAF and NIS Genes Expression in CD133pos Cells Segregated from ATC Cell Lines after Treatment with Selumetinib. BRAF mRNA levels down-regulated in CD133pos cells separated from C643 (I), SW1736 (III), and 8305C (V) cell lines, after incubation with selumetinib (10, 15, 20, 25 nM), for 24 and 48 hours, respectively. Also after treatment with these inhibitor concentrations, NIS mRNA levels up-regulated in CD133pos cells isolated from C643 (II), SW1736 (IV), 8305C (VI) cell lines after 24 and 48 hours, respectively. GAPDH gene primer was used for mRNAs levels normalization. ⃰ P<0.05 by One-way ANOVA test.

Article Snippet: Anaplastic Thyroid Carcinoma cell lines and cell culture The ATC cell line (8305C) was bought from the National Cell Bank of Iran (Pasteur Institute of Iran, Tehran, Iran).

Techniques: Expressing, Incubation, Isolation

Journal: Asian Pacific Journal of Cancer Prevention

Article Title: Targeting the BRAF Signaling Pathway in CD133pos Cancer Stem Cells of Anaplastic Thyroid Carcinoma

doi: 10.31557/apjcp.2019.20.5.1353

Figure Lengend Snippet: Figure 6. ICC Results of the CD133pos Cells which Separated from ATC Cell Lines. Left panel: CD133pos cells of C643, SW1736 and 8305C cell lines without treatment with selumetinib as well as negative control (A-C). The results showed that the NIS protein was presented in cytoplasm and plasma membrane of some CD133pos cells after treatment with Selumetinib in different concentrations and times (20 nM, 24h, D-F) and (10 nM, 48h, G-I). Magnification, × 400.

Article Snippet: Anaplastic Thyroid Carcinoma cell lines and cell culture The ATC cell line (8305C) was bought from the National Cell Bank of Iran (Pasteur Institute of Iran, Tehran, Iran).

Techniques: Negative Control, Clinical Proteomics, Membrane