Journal: International Journal of Hepatology

Article Title: Enhanced Antitumor Activity with Combining Effect of mTOR Inhibition and Microtubule Stabilization in Hepatocellular Carcinoma

doi: 10.1155/2013/103830

Figure Lengend Snippet: Everolimus inhibited proliferation and mTOR signaling in HCC cell lines. (a) Dose-dependent inhibition of HCC cell proliferation by everolimus. The effect of everolimus on cell viability was assessed by MTT assay. Dose-response curves of everolimus for all HCC cell lines were shown. Similar results were observed in 3 independent experiments. (b) Average IC 50 values of everolimus in HCC cell lines. Cumulative results from 3 independent experiments were shown as mean ± SEM. (c) Everolimus inhibited the mTOR pathway in HCC cells. HepG2, Hep3B, and SNU398 cells (3 × 10 5 ) were treated with 0.1 μ M everolimus (hereafter labeled as Eve) or DMSO control for 48 hrs and 72 hrs. The expression levels of the mTOR pathway components, pi-mTOR (ser2448), mTOR, pi-p70S6K (Thr389), p70S6K, pi-S6 (ser240/244), S6, pi-4E-BP1 (ser65), and 4E-BP1, and actin were examined by western blotting. Similar results were observed in 3 independent experiments.

Article Snippet: The following antibodies were used in the study: anti-mTOR, anti-pi-mTOR (ser2448), anti-Akt, anti-pi-Akt (ser473), anti-p70S6k, anti-pi-p70S6k (Thr389), anti-S6, anti-pi-S6 (ser240/244), anti-4E-BP1, anti-pi-4E-BP1 (ser65), anti-cleaved PARP (all from Cell Signaling Technology, Beverly, MA, USA), and anti-actin (Calbiochem, Nottingham, UK).

Techniques: Inhibition, MTT Assay, Labeling, Expressing, Western Blot

Journal: International Journal of Hepatology

Article Title: Enhanced Antitumor Activity with Combining Effect of mTOR Inhibition and Microtubule Stabilization in Hepatocellular Carcinoma

doi: 10.1155/2013/103830

Figure Lengend Snippet: Enhanced antitumor activity of the everolimus/patupilone combination in HCC cell lines. (a) Effects of everolimus/patupilone in HCC cell lines. HepG2, Hep3B, and SNU398 cells (1 × 10 4 ) were treated with various concentrations of everolimus in combination with 0.5 nM patupilone (Pat) for 24 hrs. Cell viability was assessed by MTT assay. Cumulative results from 3 independent experiments were shown as mean ± SEM (* P < 0.05, ** P < 0.01, *** P < 0.001 versus everolimus-treated group). (b) The mTOR signaling in HCC cells was not further suppressed by the everolimus/patupilone combination treatment. HepG2, Hep3B, and SNU398 cells (3 × 10 5 ) were treated with everolimus (0.1 μ M) and/or patupilone (Pat) (0.5 nM) for 24 hrs. The everolimus/patupilone combination is abbreviated as Eve/Pat hereafter. The expression levels of the mTOR pathway components, pi-mTOR (ser2448), mTOR, pi-p70S6K (Thr389), p70S6K, pi-S6 (ser240/244), S6, pi-4E-BP1 (ser65), and 4E-BP1, and actin were examined by western blotting. Similar results were observed in 3 independent experiments.

Article Snippet: The following antibodies were used in the study: anti-mTOR, anti-pi-mTOR (ser2448), anti-Akt, anti-pi-Akt (ser473), anti-p70S6k, anti-pi-p70S6k (Thr389), anti-S6, anti-pi-S6 (ser240/244), anti-4E-BP1, anti-pi-4E-BP1 (ser65), anti-cleaved PARP (all from Cell Signaling Technology, Beverly, MA, USA), and anti-actin (Calbiochem, Nottingham, UK).

Techniques: Activity Assay, MTT Assay, Expressing, Western Blot