specific shp-1 inhibitor  (Cayman Chemical)

Journal: Breast Cancer Research : BCR

Article Title: Novel sorafenib analogues induce apoptosis through SHP-1 dependent STAT3 inactivation in human breast cancer cells

doi: 10.1186/bcr3457

Figure Lengend Snippet: SHP-1-dependent inhibition of STAT3 mediates the apoptotic effects of SC-1 and SC-43 in breast cancer cells. A , inhibition of SHP-1 reverses effects of SC-1 and SC-43 on apoptosis. MDA-MB-468 cells were pretreated with 50 μM sodium vanadate, a non-specific phosphatase inhibitor (Left), or 50 μM PTP inhibitor III, a specific SHP-1 inhibitor (Right), for 60 minutes and then treated with SC-1 or SC-43 at 7.5 μM for 36 hours. B , silencing SHP-1 by siRNA reduces the effects of SC-1 and SC-43 on p-STAT3 inhibition and apoptosis in MDA-MB-453 and MDA-MB-468 cells. Cells were transfected with control siRNA (scrambled) or SHP-1 siRNA for 48 hours then treated with SC-1 or SC-43 at 7.5 μM for another 24 hours. C , effects of sorafenib, SC-1 and SC-43 on SHP-1 expression and phosphorylation (Tyr536 and Ser591). MDA-MB-468 cells were exposed to sorafenib, SC-1 or SC-43 at 7.5 μM for 36 hours. Western blot data are representative of three independent experiments. D , the activity of SHP-1 (Upper) and SHP-2 (Lower) in drug-treated MDA-MB-468 cells. Cells were treated with sorafenib, SC-1 or SC-43 at 7.5 μM for 36 hours and cell lysates were assayed for phosphatase activity as described in Methods. E , dose-escalation effects of SC-43 on the activity of SHP-1 (Upper) and SHP-2 (Lower) in drug-treated MDA-MB-468 cells. Cells were treated with SC-43 at the indicated doses for 36 hours. F , effect of SC-43 on SHP-1 activity in drug-treated HCC-1937, MDA-MB-231, MCF-7, MDA-MB-453 and SK-BR3 cells (Upper); cells were treated with SC-43 at 10 μM for 36 hours. Effects of sorafenib, SC-1 and SC-43 on phosphatase activity in recombinant SHP-1 protein (Lower). Recombinant SHP-1 protein (25 ng) was incubated with drugs at 100 nM for 30 minutes. For bar charts, Columns, mean; bars, SD ( n = 3). * P <0.05.

Article Snippet: Sodium vanadate and specific SHP-1 inhibitor were purchased from Cayman Chemical (Ann Arbor, MI, USA).

Techniques: Inhibition, Transfection, Expressing, Western Blot, Activity Assay, Recombinant, Incubation

Journal: Breast Cancer Research : BCR

Article Title: Novel sorafenib analogues induce apoptosis through SHP-1 dependent STAT3 inactivation in human breast cancer cells

doi: 10.1186/bcr3457

Figure Lengend Snippet: Therapeutic evaluation of SC-1 and SC-43 on breast cancer xenograft tumor growth in vivo . A , SC-1 and SC-43 show significant antitumor effects on MDA-MB-468 tumors. Growth curves of MDA-MB-468 tumors treated with vehicle, sorafenib, SC-1 and SC-43. Points, mean (n = 6); bars, SE. B , left, western blot analysis of p-STAT3 and STAT3 in MDA-MB-468 tumors. Right, the activity of SHP-1 in MDA-MB-468 tumors. Tumors were harvested 28 days after treatment and assayed for molecular events by western blotting and for SHP-1 activity. Columns, mean; bars, SD (n = 3). * P <0.05. C , overexpressed STAT3 in MDA-MB-468 tumors shows a significant protective effect against treatment with SC-43. Left , p-STAT3 and STAT3 expressions in STAT3-overexpressed and in empty vector- transfected MDA-MB-468 cells. Middle , growth curves of STAT3-overexpressed MDA-MB-468 tumors treated with vehicle and SC-43. Points, mean (n = 6); bars, SE. Right, western blot analysis of p-STAT3 and STAT3 in STAT3-overexpression MDA-MB-468 tumors treated with vehicle or SC-43. Tumors were harvested 28 days after treatment and assayed for molecular events by western blotting. D , body weight of xenograft mice bearing MDA-MB-468 tumors (left) and STAT3-overexpression MDA-MB-468 tumors (right) during the in vivo experiment. Points, mean (n = 6); bars, SE. Female NCr athymic nude mice (four to six weeks of age) were used for experiments (A) to (D). Mice were treated with sorafenib, SC-1 or SC-43 (10 mg/kg body weight) per os daily. Controls received vehicle. E , scheme of SHP-1 mediated drug mechanism of SC-1 and SC-43 in breast cancer cells.

Article Snippet: Sodium vanadate and specific SHP-1 inhibitor were purchased from Cayman Chemical (Ann Arbor, MI, USA).

Techniques: In Vivo, Western Blot, Activity Assay, Plasmid Preparation, Transfection, Over Expression

Journal: Breast Cancer Research : BCR

Article Title: Novel sorafenib analogues induce apoptosis through SHP-1 dependent STAT3 inactivation in human breast cancer cells

doi: 10.1186/bcr3457

Figure Lengend Snippet: Immunohistochemical stain for p-STAT3 and SHP-1 in a clinical sample. Representative immunohistochemical patterns showing (A) strong nuclear expression with no cytoplasmic expression for STAT3 in cancer cells, (B) no nuclear expression with mild cytoplasmic expression for SHP-1 in cancer cells, (C) neither nuclear expression nor cytoplasmic expression for STAT3 in adjacent normal breast tissue, and (D) no nuclear expression and strong cytoplasmic expression for SHP-1 in adjacent normal breast tissue.

Article Snippet: Sodium vanadate and specific SHP-1 inhibitor were purchased from Cayman Chemical (Ann Arbor, MI, USA).

Techniques: Immunohistochemical staining, Staining, Expressing