Journal: Cancers

Article Title: Ras Pathway Activation and MEKi Resistance Scores Predict the Efficiency of MEKi and SRCi Combination to Induce Apoptosis in Colorectal Cancer

doi: 10.3390/cancers14061451

Figure Lengend Snippet: CellTiter-Glo analysis to compare cell dose response curve for Trametinib and Dasatinib for all 16 CRC cell lines used in this study. The assays were performed 96 h post drug treatment with drug concentrations ranging from 0.01 µM–1 µM. Response to both drugs increases as dosing increases. For most of the cell lines, response plateaus at 0.1 µM concentration (showing less than 50% viability), though there are few exceptions ( n = 3).

Article Snippet: All CRC cell lines were treated with Trametinib (HY-10999) or Dasatinib (HY-10181) sourced from Medchem Express.

Techniques: Concentration Assay

Journal: Cancers

Article Title: Ras Pathway Activation and MEKi Resistance Scores Predict the Efficiency of MEKi and SRCi Combination to Induce Apoptosis in Colorectal Cancer

doi: 10.3390/cancers14061451

Figure Lengend Snippet: CellTiter-Glo analysis for cell viability percentage of all cell lines, used in this study, treated with Trametinib alone, Dasatinib alone and combination (all 0.1 µM for 96 h). Student’s t -test (paired one-tailed) demonstrates a significant decrease in cell viability with use of the drug combination in comparison to using the drugs alone for the following cell lines: LIM2099, LS123, WiDr, SW480, SNU2CA, HCT116 and HCA-7. All other cell lines show nonsignificant decreases with the use of the combination compared to single use Trametinib ( n = 3).

Article Snippet: All CRC cell lines were treated with Trametinib (HY-10999) or Dasatinib (HY-10181) sourced from Medchem Express.

Techniques: One-tailed Test, Comparison

Journal: Cancers

Article Title: Ras Pathway Activation and MEKi Resistance Scores Predict the Efficiency of MEKi and SRCi Combination to Induce Apoptosis in Colorectal Cancer

doi: 10.3390/cancers14061451

Figure Lengend Snippet: Annexin V analysis for apoptosis of cell lines treated with Trametinib, Dasatinib, and combination (all 0.1 µM for 96 h). Student’s t -test demonstrates a significant increase in (singular drug) apoptosis with use of the drug combination for cell lines LIM2099, LS123, WiDr, SNU2CA, SW480, HCA-7, SW1417, and HCT116. All other cell lines prove to have nonsignificant increases with use of the combination ( n = 3).

Article Snippet: All CRC cell lines were treated with Trametinib (HY-10999) or Dasatinib (HY-10181) sourced from Medchem Express.

Techniques:

Journal: Cancers

Article Title: Ras Pathway Activation and MEKi Resistance Scores Predict the Efficiency of MEKi and SRCi Combination to Induce Apoptosis in Colorectal Cancer

doi: 10.3390/cancers14061451

Figure Lengend Snippet: Western Blot analysis of cell lines treated with Trametinib, Dasatinib or combination. Cleaved PARP, Phospho-SRC (Tyr416), SRC, Phospho-ERK1/2 (Thr202/Tyr204), and ERK were probed for. This analysis shows increased cleaved PARP in cells shown to be sensitive with the combination treatment in Annexin V analysis (fig4). Increases in cleaved PARP is shown in all the sensitive cell lines. Inhibitory effects of the drugs are also confirmed with decrease of Phospho-ERK and Phospho-SRC levels in treated cell lines. However, notably some resistant cell lines did not show significant decreases in P-SRC when Dasatinib was used. All findings were confirmed via band density readings featured in the histograms.

Article Snippet: All CRC cell lines were treated with Trametinib (HY-10999) or Dasatinib (HY-10181) sourced from Medchem Express.

Techniques: Western Blot

Journal: Cancers

Article Title: Ras Pathway Activation and MEKi Resistance Scores Predict the Efficiency of MEKi and SRCi Combination to Induce Apoptosis in Colorectal Cancer

doi: 10.3390/cancers14061451

Figure Lengend Snippet: The 18-gene RAS activation score and the 13-gene MEKi Resistance score for the 16 CRC cell lines used in this study were plotted with color indicators for their drug sensitivity. Each cell line is represented as a singular dot that is colored, indicating its apoptotic sensitivity to either Trametinib, Dasatinib, or the combination of both. Cells with the checkerboard pattern are cells with significantly enhanced apoptosis when the combination is used. Notably, 7 out of 9 of the CMS4 cell lines have increased cell death when the combination is used. These cells all have positive 18-gene and 13-gene scores (located in the righthand upper quadrant).

Article Snippet: All CRC cell lines were treated with Trametinib (HY-10999) or Dasatinib (HY-10181) sourced from Medchem Express.

Techniques: Activation Assay

Journal: Scientific reports

Article Title: Melanoma cells adopt features of both mesenchymal and amoeboid migration within confining channels.

doi: 10.1038/s41598-021-97348-7

Figure Lengend Snippet: Figure 3. Pharmacologically inhibiting focal adhesion formation increases the speed and directionality of blebbing cells. (A) Confirmation that a melanoma A375-M2 cell with the focal adhesion marker, EGFP- paxillin, within a fibronectin coated (10 µg/mL) microchannel does not form focal adhesions after treatment with Dasatinib (0.5 µM). Middle (left) and bottom (right) focal planes are shown. (B) Montage of a melanoma A375-M2 cell treated with Src family kinase inhibitor, Dasatinib (0.5 µM). Cells were visualized using a far red membrane dye. Zoom shows prominent blebs at the cell leading edge. (C) A comparison of instantaneous speeds for phenotype 1 (N = 28), 2 (N = 31), and Dasatinib (N = 75) treated cells. Statistical significance was determined by a Dunn’s multiple-comparison test post-hoc. (D) A comparison of directionality ratio over time for phenotype 1 (N = 28), 2 (N = 31), and Dasatinib (N = 75) treated cells. Error is SEM. (E) A comparison of Mean Square Displacement (MSD; square microns) over time for phenotype 1 (N = 28), 2 (N = 31), and Dasatinib (N = 75) treated cells. As noted on the right of the graph, a sharp decrease in the graph is the result of fast cells leaving the channels. All data are representative of at least three independent experiments. *—p ≤ 0.05, **—p ≤ 0.01, ***—p ≤ 0.001, and ****—p ≤ 0.0001.

Article Snippet: Dasatinib (cat no. 6793) and CK-666 (cat no. 3950) were purchased from Tocris Bioscience (Bristol, UK).

Techniques: Marker, Membrane, Comparison

Journal: Molecular biomedicine

Article Title: Integrin-Src-YAP1 signaling mediates the melanoma acquired resistance to MAPK and PI3K/mTOR dual targeted therapy.

doi: 10.1186/s43556-020-00013-0

Figure Lengend Snippet: Fig. 5 The inhibition of Src suppressed the growth of dual-drug resistant cells. a SKMEL28 DPR and WM2664 DPR were transduced with shRNA targeting nontarget (vector) or shRNA targeting against integrin α3, α11 and β1. Cell lysates were made for immunoblot analysis with antibodies indicated. β-ACTIN is as loading control. b Survival curves of SKMEL28 DPR and WM2664 DPR titrated with dasatinib for 72 h. Results are shown relative to DMSO-treated controls (mean ± SEM, n = 5; dashed line, 50% inhibition). c Long-term colony assays of resistant sub-lines treated with dasatinib as indicated concentration. d Quantification of (c) by Image J, ∗p < 0.05, ∗∗p < 0.01, or ∗∗∗p < 0.001. e Clonogenic assays of DPRs engineered control (vector) or Src-targeting shRNA treated with 1 μM AZD6244 and 0.1 μM BEZ235. f Survival curves of resistant sublines of SKME L28 and WM2664 titrated with AZD6244 + BEZ235 with or without PF-562271 for 72 h. Results are shown relative to DMSO-treated controls (mean ± SEM, n = 5; dashed line, 50% inhibition). g Long-term colony assays of SKMEL28 DPR, WM2664 TPR and WM2664 DPR treated with PF- 562271. h Quantification of (g) by Image J, ∗p < 0.05, ∗∗p < 0.01, or ∗∗∗p < 0.001

Article Snippet: AZD6244 (MEKi), BEZ235 (PI3K/mTORi), CCG-1423 (RhoAi), QNZ (EVP4593) (NF-κBi), C188–9 (STAT3i), dasatinib (Srci), verteporfin (YAP1i), XAV-939 (WNTi), SB525334 (TGFβi), doramapimod (p38i), Defactinib (FAKi), and PF-562271 (FAKi) were purchased from SELLECK.

Techniques: Inhibition, Transduction, shRNA, Plasmid Preparation, Western Blot, Control, Concentration Assay

Journal: Journal of Hematology & Oncology

Article Title: Mouse avatar models of esophageal squamous cell carcinoma proved the potential for EGFR-TKI afatinib and uncovered Src family kinases involved in acquired resistance

doi: 10.1186/s13045-018-0651-z

Figure Lengend Snippet: Increased phosphorylation of Src family kinases leads to acquired afatinib resistance in ESCC. a Increased pSFKs levels were observed in KYSE450-R and PDX03-R resistant models, but total SFKs levels were unchanged. Cells were harvested after treatment with 200 nM afatinib for 48 h. The PDX lysates used were the same as those described in Fig. d. All assays were repeated three times independently. b Resistant cells were treated with the indicated concentrations of afatinib in the presence or absence of 100 nM dasatinib for 72 h, and CCK-8 assays were performed to assess cell viability. Data are presented as the means ± SDs of three independent assays. c KYSE450-R and EC109-R cells were treated with 200 nM afatinib alone or in combination with 100 nM dasatinib for 48 h. d , e Curves showing the xenografts growth of KYSE450-R ( d ) and PDX03-R ( e ) treated with vehicle control, afatinib (15 mg/kg), crizotinib (25 mg/kg), afatinib (15 mg/kg) plus crizotinib (25 mg/kg), dasatinib (15 mg/kg), or afatinib (15 mg/kg) plus dasatinib (15 mg/kg). Data are presented as means ± SDs; n = 5. Mice were sacrificed after 21 days of treatment, and xenografts were isolated. Pictures of the xenografts are shown with the corresponding TGI listed in the tables. f , g Lysates were extracted from KYSE450-R ( f ) and PDX03-R ( g ) xenografts after 21 days of treatment with the corresponding inhibitors and analyzed by western blotting to explore the downstream signaling responses. The lysates were then probed with the indicated antibodies. All experiments were repeated three times independently. h A schematic of the molecular mechanisms of acquired resistance revealed in this study

Article Snippet: Gefitinib (#S1025), osimertinib (#S7297), dasatinib hydrochloride (#HY-10181A), and crizotinib hydrochloride (#HY-50878A) were purchased from Selleck Chemicals or MedChem Express.

Techniques: Phospho-proteomics, CCK-8 Assay, Control, Isolation, Western Blot

Journal: International journal of cancer

Article Title: SRC inhibition represents a potential therapeutic strategy in liposarcoma.

doi: 10.1002/ijc.29645

Figure Lengend Snippet: Figure 2. (a) SRC siRNA-mediated knockdown (confirmed by Western blots) and (b) Dasatinib treatment lead to a significant decrease in cellular viability of all tested LS cell lines in MTT assays. (c) Coincubation of Dasatinib (20 nM) with increasing concentrations of Doxorubicin (DOXO), Vincristin (VCR) and Actinomycin-D (ACT-D) resulted in additive effects. Results were considered significant with p-values ***<0.001; **<0.01; *<0.05.

Article Snippet: Inhibitor treatment was performed with increasing concentrations of Dasatinib (Santa Cruz Biotech, Heidelberg, Germany) and DMSO as control.

Techniques: Knockdown, Western Blot

Journal: International journal of cancer

Article Title: SRC inhibition represents a potential therapeutic strategy in liposarcoma.

doi: 10.1002/ijc.29645

Figure Lengend Snippet: Figure 3. Western blot analyses revealed dose-dependent dephosphorylation of p-(Tyr416)-SRC and its interaction partners in LS cells upon treatment with increasing concentrations of Dasatinib (0.03–10 mM).

Article Snippet: Inhibitor treatment was performed with increasing concentrations of Dasatinib (Santa Cruz Biotech, Heidelberg, Germany) and DMSO as control.

Techniques: Western Blot, De-Phosphorylation Assay

Journal: International journal of cancer

Article Title: SRC inhibition represents a potential therapeutic strategy in liposarcoma.

doi: 10.1002/ijc.29645

Figure Lengend Snippet: Figure 4. Mitotic and apoptotic rates in LS cells treated with Dasatinib. (a) Representative results of flow cytometric analyses of p-(Ser10)- histone H3 and cleaved PARP (Asp214) in LS cells, incubated with 1 mM Dasatinib or DMSO as control. (b) Induction of apoptosis was inde- pendently assessed by DAPI staining of LS cell lines treated with Dasatinib (0.1–1.0 mM). For statistical analysis, unpaired Student’s t-tests were performed (***p < 0.001; **p < 0.01). [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]

Article Snippet: Inhibitor treatment was performed with increasing concentrations of Dasatinib (Santa Cruz Biotech, Heidelberg, Germany) and DMSO as control.

Techniques: Incubation, Control, Staining

Journal: International journal of cancer

Article Title: SRC inhibition represents a potential therapeutic strategy in liposarcoma.

doi: 10.1002/ijc.29645

Figure Lengend Snippet: Figure 5. Investigation of LS motility and invasive potential upon SRC inhibition. (a) Representative wound scratch assays in MLS and PLS cells treated with 0.01 mM Dasatinib or DMSO as control. (b) Transwell chamber and (c) Matrigel chamber assays of LS cell lines incubated with different concentrations of Dasatinib (0.01–0.03 mM) for 24 hr. For statistical analysis, unpaired Student’s t-tests were performed (***p < 0.001; **p < 0.01; *p < 0.05).

Article Snippet: Inhibitor treatment was performed with increasing concentrations of Dasatinib (Santa Cruz Biotech, Heidelberg, Germany) and DMSO as control.

Techniques: Inhibition, Control, Incubation

Journal: Toxicology Reports

Article Title: Dasatinib targets c-Src kinase in cardiotoxicity

doi: 10.1016/j.toxrep.2023.04.013

Figure Lengend Snippet: Toxicity of dasatinib on resident cardiac cells A. Neonatal rat ventricular cardiomyocytes were treated with different concentrations of dasatinib for 24 h. Shown are analysis for cardiomyocyte viability by ATP assay (left panel) and analysis for cell death by measuring the release of adenylate kinase from ruptured cardiomyocytes (right panel). N = 6. B. Human umbilical vein endothelial cells treated with dasatinib for 24 h. Shown is analysis for cell viability by ATP assay and analysis for cell death by measuring the release of adenylate kinase. N = 6. C. Human cardiac fibroblasts treated with dasatinib for 24 h. Shown is analysis for cell viability by ATP assay and analysis for cell death by measuring the release of adenylate kinase. N = 6. *p < 0.05, * *p < 0.01, * **p < 0.001 vs Vehicle. Statistics were calculated by one-way analysis of variance followed by Dunnett’s post hoc test.

Article Snippet: Wild type c-Src (pBABE-Src-Rescue, Addgene plasmid # 26983; http://n2t.net/addgene:26983; RRID:Addgene_26983) and dasatinib resistant c-Src (pBABE-Src-Dasatinib-resistant, Addgene plasmid # 26980; http://n2t.net/addgene:26980; RRID:Addgene_26980) were a gift from Joan Massague.

Techniques: ATP Assay

Journal: Toxicology Reports

Article Title: Dasatinib targets c-Src kinase in cardiotoxicity

doi: 10.1016/j.toxrep.2023.04.013

Figure Lengend Snippet: Analysis for target of dasatinib in cardiomyocytes A. Neonatal rat cardiomyocytes were treated with 100 nM and 300 nM dasatinib for 24 h and co-treated with fibroblast growth factor (FGF) where indicated. Shown is Western blot analysis for phosphorylated STAT3, Akt and ERK. GAPHD was used as a loading control. B. Dose response curve of the cardiomyocyte toxicity of dasatinib. Cardiomyocytes were treated with 10, 30, 100, 300 and 1000 nM dasatinib for 24 h and data from ATP assay were plotted against dasatinib concentration. C. Profiling data of kinases targeted by dasatinib.

Article Snippet: Wild type c-Src (pBABE-Src-Rescue, Addgene plasmid # 26983; http://n2t.net/addgene:26983; RRID:Addgene_26983) and dasatinib resistant c-Src (pBABE-Src-Dasatinib-resistant, Addgene plasmid # 26980; http://n2t.net/addgene:26980; RRID:Addgene_26980) were a gift from Joan Massague.

Techniques: Western Blot, Control, ATP Assay, Concentration Assay

Journal: Toxicology Reports

Article Title: Dasatinib targets c-Src kinase in cardiotoxicity

doi: 10.1016/j.toxrep.2023.04.013

Figure Lengend Snippet: Gatekeeper mutation in c-Src rescues the cardiomyocytes from dasatinib-induced toxicity Neonatal rat ventricular cardiomyocytes were transfected with wild type c-Src or with c-Src T338I. A. Shown is Western blot analysis for c-Src and GAPDH. B. Shown is analysis for cardiomyocyte viability by ATP assay after treatment of cells with 100 nM or 300 nM dasatinib for 24 h. N = 4. * *p < 0.01, * **p < 0.001 vs Vehicle. Statistics were calculated by one-way analysis of variance followed by Dunnett’s post hoc test. C. Data shown as a comparison of toxicity of dasatinib on control cardiomyocytes and cardiomyocytes transfected with wild type c-Src or c-Src T338I. N = 4. #p < 0.05 vs control cells treated with indicated dose of dasatinib. Statistics were calculated by one-way analysis of variance followed by Tukey’s post hoc test.

Article Snippet: Wild type c-Src (pBABE-Src-Rescue, Addgene plasmid # 26983; http://n2t.net/addgene:26983; RRID:Addgene_26983) and dasatinib resistant c-Src (pBABE-Src-Dasatinib-resistant, Addgene plasmid # 26980; http://n2t.net/addgene:26980; RRID:Addgene_26980) were a gift from Joan Massague.

Techniques: Mutagenesis, Transfection, Western Blot, ATP Assay, Comparison, Control