Journal: BMC Cancer

Article Title: Prognostic significance and therapeutic potential of the activation of anaplastic lymphoma kinase/protein kinase B/mammalian target of rapamycin signaling pathway in anaplastic large cell lymphoma

doi: 10.1186/1471-2407-13-471

Figure Lengend Snippet: Histology and immunohistochemistry (IHC) of ALCL tumor samples. (A, B) Histology of ALCL tumor samples (HE staining). (A) The classic type of ALCL. The tumor cells are large with abundant cytoplasm and manifest prominent nucleoli with an eccentrically located and pleomorphic nucleus of kidney-shape. (B) ALCL of small cell type. The tumor cells are small in size, with abundant cytoplasm and prominent nucleoli. (C to H) IHC staining of ALCL tumor samples (EnVision staining). Expression of ALK, p-AKT, p-mTOR, p-4E-BP1, and p-p70S6K eIF4E was assessed. (C, D) IHC staining with ALK1 antibody. (C) The tumor cells show cytoplasmic and nuclear staining. (D) The tumor cells manifest cytoplasmic staining only. (E) IHC staining with Thr308p-AKT antibody. The tumor cells show cytoplasmic and nuclear staining. (F) IHC staining with Ser2448p-mTOR antibody. The tumor cells show cytoplasmic staining. (G) IHC staining with Thr70p-4E-BP1 antibody. The tumor cells show cytoplasmic staining. (H) IHC staining with Thr421p-p70S6K antibody. The tumor cells show cytoplasmic staining. All images were captured at 400× magnification.

Article Snippet: Rabbit polyclonal antibodies specific for Thr308p-AKT (p-AKT), Ser2448p-mTOR (p-mTOR), Thr70p-4E-BP1 (p-4E-BP1), and Thr421p-p70S6K1 (p-p70S6K1) (Cell Signaling Technology, Beverly, MA) were used.

Techniques: Immunohistochemistry, Staining, Expressing

Journal: BMC Cancer

Article Title: Prognostic significance and therapeutic potential of the activation of anaplastic lymphoma kinase/protein kinase B/mammalian target of rapamycin signaling pathway in anaplastic large cell lymphoma

doi: 10.1186/1471-2407-13-471

Figure Lengend Snippet: Relationship between expression of p-4E-BP1 and p-p70S6K and activation of ALK, AKT, and mTOR

Article Snippet: Rabbit polyclonal antibodies specific for Thr308p-AKT (p-AKT), Ser2448p-mTOR (p-mTOR), Thr70p-4E-BP1 (p-4E-BP1), and Thr421p-p70S6K1 (p-p70S6K1) (Cell Signaling Technology, Beverly, MA) were used.

Techniques: Expressing, Activation Assay, Significance Assay

Journal: BMC Cancer

Article Title: Prognostic significance and therapeutic potential of the activation of anaplastic lymphoma kinase/protein kinase B/mammalian target of rapamycin signaling pathway in anaplastic large cell lymphoma

doi: 10.1186/1471-2407-13-471

Figure Lengend Snippet: Overexpression of NPM-ALK activated the AKT/mTOR pathway. Overexpression of NPM-ALK in BaF3 cells induced hyper-activation of the AKT/mTOR pathway, as demonstrated by hyperphosphorylation of AKT and downstream molecules of mTOR: 4E-BP1 and p70SK6.

Article Snippet: Rabbit polyclonal antibodies specific for Thr308p-AKT (p-AKT), Ser2448p-mTOR (p-mTOR), Thr70p-4E-BP1 (p-4E-BP1), and Thr421p-p70S6K1 (p-p70S6K1) (Cell Signaling Technology, Beverly, MA) were used.

Techniques: Over Expression, Activation Assay

Journal: International Journal of Oncology

Article Title: Theaflavin-3, 3′-digallate decreases human ovarian carcinoma OVCAR-3 cell-induced angiogenesis via Akt and Notch-1 pathways, not via MAPK pathways

doi: 10.3892/ijo.2015.3257

Figure Lengend Snippet: Angiogenesis-related proteins are affected by TF3 treatment in OVCAR-3 cells. Western blot analysis revealed that TF3 decreased the protein level of p-Akt, p-mTOR, p-p70S6K, p-4E-BP1, Notch-1 (NICD), c-Myc and HIF-1α in OVCAR-3 cells. TF3 had no impact on the protein level of p-ERK1/2, ERK1/2, JNK, p38 and p-FoxO 1. GAPDH served as the loading control.

Article Snippet: Antibodies against phosphor-4E-BP1 (Ser65/Thr70) (p-4E-BP1), c-Myc, phosphor-extracellular signal-regulated kinases 1/2 (Thr202/Tyr204) (p-ERK1/2), ERK1/2 and GAPDH were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz).

Techniques: Western Blot

Journal: International Journal of Oncology

Article Title: Theaflavin-3, 3′-digallate decreases human ovarian carcinoma OVCAR-3 cell-induced angiogenesis via Akt and Notch-1 pathways, not via MAPK pathways

doi: 10.3892/ijo.2015.3257

Figure Lengend Snippet: Akt/mTOR/p70S6k/4E-BP1 pathway and Akt/c-Myc pathway are involved in TF3-induced inhibition of HIF-1α and VEGF. (A) Western blot analysis showed that 100 nM wortmannin, 10 μM TF3 and 100 nM wortmannin+10 μM TF3 decreased the phosphorylation of Akt, mTOR, p70S6K and 4E-BP1, and expression of c-Myc and HIF-1α. TF3+wortmannin exhibited the strongest effect among them. GAPDH served as the loading control. (B) Luciferase reporter assay and (C) VEGF ELISA showed 100 nM wortmannin, 10 μM TF3 and 100 nM wortmannin+10 μM TF3 suppressed the transcriptional activity of VEGF promoter and VEGF secretion, respectively. TF3+wortmannin elicited strongest effect among them. (D) Overexpression of active Akt attenuated the 15 μM TF3-induced decrease of phosphorylation of Akt, mTOR, p70S6K and 4E-BP1, and expression of c-Myc and HIF-1α. Overexpression of Akt, mTOR, p70S6K or 4E-BP1 attenuated TF3-induced inhibition of transcriptional activity of VEGF promoter (E) and HIF-1α promoter (F). (G) Overexpression of active Akt reversed the 15 μM TF3-induced reduction of VEGF secretion. The data are presented as the mean ± standard error of mean. * P<0.05 compared with control or between specific groups.

Article Snippet: Antibodies against phosphor-4E-BP1 (Ser65/Thr70) (p-4E-BP1), c-Myc, phosphor-extracellular signal-regulated kinases 1/2 (Thr202/Tyr204) (p-ERK1/2), ERK1/2 and GAPDH were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz).

Techniques: Inhibition, Western Blot, Expressing, Luciferase, Reporter Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Over Expression

Journal: PLoS ONE

Article Title: Utility of a PI3K/mTOR Inhibitor (NVP-BEZ235) for Thyroid Cancer Therapy

doi: 10.1371/journal.pone.0046726

Figure Lengend Snippet: p-AKT (Thr308) and p-AKT (Ser473) was increased in 8505C for more than 24 hours. p-AKT (Thr308) and p-AKT (Ser473) was decreased transiently in TT and BHP7-13. p-4E-BP1 (Thr70), p-4E-BP1 (Thr37/46) and p-S6 ribosomal protein (Ser235/236) were consistently reduced from 2 hours through over 24 hours in three cell lines. BEZ235 caused a rapid increase of p- ERK1/2 (Thr202/Tyr204) by 2 to 4 hours in three cell lines.

Article Snippet: Antibodies targeting p-AKT (Thr308), p-AKT (Ser473), AKT, p-4E-BP1 (Thr70), p-4E-BP1 (Thr37/46), 4E-BP1, p-S6 ribosomal proteins (Ser235/236), p-S6 ribosomal protein (Ser240/244), S6 ribosomal protein, p-ERK1/2 (Thr202/Tyr204), ERK1/2, p53, p21, p27, cyclin D1, caspase-3 and proliferating cell nuclear antigen (PCNA) were from Cell Signaling Technology. α-tubulin antibody was from Sigma.

Techniques: