tyrosine kinase inhibitor sunitinib malate  (MultiTarget Pharmaceuticals)

Journal: Cell Death & Disease

Article Title: Novel engineered, membrane-localized variants of vascular endothelial growth factor (VEGF) protect retinal ganglion cells: a proof-of-concept study

doi: 10.1038/s41419-018-1049-0

Figure Lengend Snippet: a Representative immunocytochemistry showing VEGFR2 (red) expression and colocalization with eVEGF-38, eVEGF-53, VEGF189 (anti-Myc tag, green), or GFP (anti-GFP, green) on the cell bodies, neurites and axons of primary RGC at DIV 3. Note the lower levels of VEGFR2 expression and the lack of long neurites and axons in the RGC-expressing GFP. Scale bar = 20 µm. b Left, representative immunocytochemical localization of eVEGF-38, eVEGF-53, and VEGF189 (anti-Myc tag, green) outlining neurites and axons of RGC, in the presence or absence of the VEGFR2 inhibitor Sunitinib malate (1 μM) or the PI3K/AKT inhibitor LY294002 (LY, 10 μM), 3 days after AAV transduction. Scale bar = 20 µm, UT = untreated. Right, quantification of total neurite length per RGC in the presence or absence of Sunitnib malate or LY294002. *** P < 0.001 compared with the corresponding untreated (UT) control, unpaired two-tailed t test, n = 15; five different fields from three independent experiments. Data = mean ± SEM. All RGC were isolated from P3 mice

Article Snippet: To determine the roles of VEGFR2 activation and PI3K/Akt signaling in neurite outgrowth, RGC were treated with the VEGFR2-selective small-molecule receptor tyrosine kinase inhibitor sunitinib malate (1 μM, Selleck Chemicals, Houston, TX) or the PI3K small-molecule inhibitor LY294002 (1 μM, Cell Signaling Technology) at DIV 2 for 24 h. Apoptotic cells were detected using the In Situ Cell Death Detection Kit (TUNEL, Roche) according to manufacturer’s instructions.

Techniques: Immunocytochemistry, Expressing, Transduction, Control, Two Tailed Test, Isolation

Journal: Human molecular genetics

Article Title: High-throughput screening uncovers miRNAs enhancing glioblastoma cell susceptibility to tyrosine kinase inhibitors.

doi: 10.1093/hmg/ddx323

Figure Lengend Snippet: Figure 1. MiR-302/520 miRNA family increases susceptibility of GBM cells to sunitinib treatment. a) Overview

Article Snippet: Temozolomide and the multitargeted tyrosine kinase inhibitors sunitinib malate and axitinib were acquired from Selleckchem (Houston, USA) and stored at 20 C in DMSO.

Techniques:

Journal: Human molecular genetics

Article Title: High-throughput screening uncovers miRNAs enhancing glioblastoma cell susceptibility to tyrosine kinase inhibitors.

doi: 10.1093/hmg/ddx323

Figure Lengend Snippet: Figure 3. Combination of miRNA-302a/520b expression with multitargeted tyrosine kinase inhibitors decreases GBM cell viability. U87 and DBTRG cells were transfected with 50 nM of miRNA-302a, miR-520b or

Article Snippet: Temozolomide and the multitargeted tyrosine kinase inhibitors sunitinib malate and axitinib were acquired from Selleckchem (Houston, USA) and stored at 20 C in DMSO.

Techniques: Expressing, Transfection

Journal: Human molecular genetics

Article Title: High-throughput screening uncovers miRNAs enhancing glioblastoma cell susceptibility to tyrosine kinase inhibitors.

doi: 10.1093/hmg/ddx323

Figure Lengend Snippet: Figure 5. Cellular DNA content increases upon miR-302a transfection and treatment with multitargeted tyrosine kinase inhibitors. Cells were incubated with miR-302a or control miRNA (cel-miR-239b) mimics for

Article Snippet: Temozolomide and the multitargeted tyrosine kinase inhibitors sunitinib malate and axitinib were acquired from Selleckchem (Houston, USA) and stored at 20 C in DMSO.

Techniques: Transfection, Incubation, Control

Journal: Cancer Biology & Therapy

Article Title: Anti-angiogenic treatment promotes triple-negative breast cancer invasion via vasculogenic mimicry

doi: 10.1080/15384047.2017.1294288

Figure Lengend Snippet: Discontinuation of sunitinib promotes TNBC tumor regrowth and invasion. (A) Effects of sunitinib on the tumor volume of TNBC and non-TNBC cells. There wa sno significant difference in tumor volume when the treatment started. After mice were treated with sunitinib for one week, both the TNBC MDA-MB-231 tumors and the non-TNBC MCF-7 tumors in the treatment groups were smaller than those in the control groups. The sunitinib-treated TNBC MDA-MB-231 tumors regrew after treatment discontinuation, while the discontinuation had no significant effect on the tumor volume of the non-TNBC MCF-7 tumors. (B) H&E staining indicated that the TNBC MDA-MB-231 tumor cells invaded into adipose tissue (arrows) and skeletal muscle tissue (arrow heads) after treatment discontinuation. There was no aggressive behavior in the non-TNBC MCF-7 tumors. (C) IHC for MMP2 staining shows that TNBC MDA-MB-231 tumors after treatment was stopped expressed a higher level of MMP2 than tumors during the sunitinib treatment. There was nodifferenceinMMP2 expression in the non-TNBC MCF-7 tumors between the sunitinib treatment and after sunitinib was discontinued. (D) Quantification of MMP2 expression in the different groups. The scale bar represents 100 μm, and the error bar indicates the standard deviation (SD). *P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The VEGF receptor tyrosine kinase inhibitor sunitinib malate (S-8803) was purchased from LC Laboratories (Boston, USA).

Techniques: Control, Staining, Expressing, Standard Deviation

Journal: Cancer Biology & Therapy

Article Title: Anti-angiogenic treatment promotes triple-negative breast cancer invasion via vasculogenic mimicry

doi: 10.1080/15384047.2017.1294288

Figure Lengend Snippet: Effects of sunitinib treatment on the expression of EMT-associated transcription factors in TNBC tumors. (A) IHC staining for VE-cadherin, Twist1, Snail and Slug. VE-cadherin is upregulated in the sunitinib-treated MDA-MB-231 tumors compared with the placebo-treated MDA-MB-231 tumors. Sunitinib increased Twist1 expression in the MDA-MB-231 tumors, and Twist1 localized to the nucleus in the treatment group. There was no change observed in the MCF-7 groups. Sunitinib did not affect the expression of Snail and Slug in the TNBC MDA-MB-231 tumors and the non-TNBC MCF-7 tumors. (B) Quantification of VE-cadherin expression in the different groups. (C)Quantification of Twist1 expression in the different groups. (D) Quantification of Slug expression in the different groups. The scale bar indicates 100 μm, and the error bar indicates the standard deviation (SD). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The VEGF receptor tyrosine kinase inhibitor sunitinib malate (S-8803) was purchased from LC Laboratories (Boston, USA).

Techniques: Expressing, Immunohistochemistry, Standard Deviation

Journal: Cancer Biology & Therapy

Article Title: Anti-angiogenic treatment promotes triple-negative breast cancer invasion via vasculogenic mimicry

doi: 10.1080/15384047.2017.1294288

Figure Lengend Snippet: Effects of sunitinib treatment on the microcirculation patterns in TNBC tumors. (A) PAS and endomucin double staining in TNBCMDA-MB-231 and non-TNBC MCF-7 tumors. There were VM channels formed by PAS-positive molecules and tumor cells in the human TNBCMDA-MB-231 tumors, whereas no VM channels were observed in the human non-TNBC MCF-7 tumors. Growth of endothelium-dependent vessels in the TNBC MDA-MB-231 tumors was blocked by sunitinib, during which the number of VM channels significantly increased. Endothelium-dependent vessel growth rebounded in MDA-MB-231 tumors after treatment discontinuation, but there was no significant difference in the number of endothelium-dependent vessels between sunitinib-treated and post-treatment non-TNBC MCF-7 tumors. (B)Quantification of VM channels in the different groups. (C)Quantification of endothelium-dependent vessels in the different groups. (D)Hypoxia and endomucin double staining inTNBCMDA-MB-231 and non-TNBC MCF-7 tumors. There were more hypoxic regions in the sunitinib-treatedMDA-MB-231 tumors compared with those in the other groups when the endothelium-dependent vessels are inhibited. After treatment discontinuation, the hypoxic area in the MDA-MB-231 tumors decreased after the endothelium-dependent vessels rebounded. The hypoxic area in the MCF-7 tumors was similar in the sunitinib-treated and discontinued groups. (E)Quantification of the hypoxic area in the different groups. The scale bar indicates 100 μm, and the error bar indicates the standard deviation (SD). * P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The VEGF receptor tyrosine kinase inhibitor sunitinib malate (S-8803) was purchased from LC Laboratories (Boston, USA).

Techniques: Double Staining, Standard Deviation

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Antiangiogenic agents increase breast cancer stem cells via the generation of tumor hypoxia

doi: 10.1073/pnas.1018866109

Figure Lengend Snippet: Sunitinib malate induces hypoxia in breast tumors in vivo and results in an increase in the tumor stem/progenitor cell population. (A) MDA-MB-231 or SUM159 cells were injected into the inguinal fat pads of NOD/SCID mice. One day after injection, mice were given either vehicle (group A) or sunitinib (60 mg/kg) (group B). Group C received the vehicle control until tumors reached an average size of 4 mm in diameter, and then were administered sunitinib (60 mg/kg) daily. Data are shown as averages ± SD. n = 8–10 (MDA-MB-231); n = 4–6 (SUM159). Treated tumors were significantly smaller than control tumors at end point. *P < 0.01. (B) CD31 staining of blood vessels (green) and DAPI nuclear staining (blue) of tumors from a control mouse and a sunitinib-treated mouse. (Scale bars, 200 μm.) (C) Representative tumors displaying a lack of vasculature in sunitinib-treated mice compared with control tumors. (D) The percentage of ALDH+ cells in tumors was determined by Aldefluor assay. n = 8. *P < 0.05. (E) Serial dilutions of cells obtained from primary SUM159 tumors treated with control or sunitinib were implanted in secondary NOD/SCID mice. Primary tumors treated with sunitinib grew secondary tumors more rapidly than the control tumors. Data are shown as averages ± SD. n = 4–5. *P < 0.05.

Article Snippet: To determine whether antiangiogenic agents stimulate an increase in breast CSCs in vivo, we treated tumor-bearing mice with the multireceptor tyrosine kinase inhibitor sunitinib malate (Sutent; Pfizer).

Techniques: In Vivo, Injection, Control, Staining

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Antiangiogenic agents increase breast cancer stem cells via the generation of tumor hypoxia

doi: 10.1073/pnas.1018866109

Figure Lengend Snippet: Sunitinib malate induces hypoxia in breast tumors in vivo, and ALDH1+ cells are concentrated in hypoxic regions. (A) Hypoxia in SUM159 tumors was detected by immunofluorescence staining of pimonidazole adducts in sections from control or sunitinib-treated animals. Staining shows pimonidazole immunodetection (green). (Scale bars, 800 μm.) (B) Staining shows pimonidazole (green) and ALDH1 (red) merged with DAPI-stained nuclei (blue). (Scale bars, 400 μm.) (Inset) Magnification: 5×. (C) Quantitation of ALDH1-positive cells in control tumors versus hypoxic and normoxic areas within sunitinib-treated tumors. Data are shown as averages ± SD. n = 5. *P < 0.05, **P < 0.01.

Article Snippet: To determine whether antiangiogenic agents stimulate an increase in breast CSCs in vivo, we treated tumor-bearing mice with the multireceptor tyrosine kinase inhibitor sunitinib malate (Sutent; Pfizer).

Techniques: In Vivo, Immunofluorescence, Staining, Control, Immunodetection, Quantitation Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Antiangiogenic agents increase breast cancer stem cells via the generation of tumor hypoxia

doi: 10.1073/pnas.1018866109

Figure Lengend Snippet: The β-catenin pathway is stimulated in response to hypoxia. (A) SUM159 and MCF-7 cells were grown under 1% (H) or 21% (N) O2 for 24 h. Immunoblotting was carried out for total and phospho-Akt and total and phospho-β-catenin. (B) SUM159 cells infected with the pGreenFire LEF/TCF lentivirus reporter were sorted by flow cytometry into GFP+ and GFP− populations, grown under 1% or 21% O2 for 20 h, and assayed for luciferase activity. GFP− cells displayed significantly higher luciferase activity in response to hypoxia. Data are shown as averages ± SD. n = 6. *P < 0.01. (C) Immunoblot for phospho-β-catenin in SUM159 cell extracts following transfection with control siRNA or HIF-1α siRNA and 3-d hypoxia treatment. (D) Representative immunofluorescent staining for β-catenin in SUM159 xenografts from control or sunitinib-treated mice. Note the primarily cytoplasmic staining in cells from control tumors and intense nuclear staining in cells near necrotic regions (N) of sunitinib-treated tumors. (Scale bars, 100 μm.)

Article Snippet: To determine whether antiangiogenic agents stimulate an increase in breast CSCs in vivo, we treated tumor-bearing mice with the multireceptor tyrosine kinase inhibitor sunitinib malate (Sutent; Pfizer).

Techniques: Western Blot, Infection, Flow Cytometry, Luciferase, Activity Assay, Transfection, Control, Staining