Journal: American Journal of Cancer Research

Article Title: Duloxetine enhances the sensitivity of non-small cell lung cancer cells to EGFR inhibitors by REDD1-induced mTORC1/S6K1 suppression

doi: 10.62347/WMQV6643

Figure Lengend Snippet: Duloxetine, but not milnacipran, venlafaxine, or desvenlafaxine, sensitizes NSCLC cells to lapatinib. A-C. H1299, H460, and A549 cells were treated with 5 μM lapatinib and the indicated concentrations of duloxetine, milnacipran, venlafaxine or desvenlafaxine for 48 h. D. H1299, H460, and A549 cells were treated with 5 μM lapatinib and/or 10 μM duloxetine for 48 h. A-C. Cell viability was measured by MTT assay. The data are presented as the mean percentage of control ± SD (n=3). D. Cell death was detected as the percentage of Annexin V and/or Propidium Iodide (PI) positive cells. DLX: duloxetine, LPT: lapatinib, PI: Propidium Iodide.

Article Snippet: Milnacipran (#S3140, purity: 99.99%), venlafaxine (#S1441, purity: 99.98%), desvenlafaxine (#S4113, purity: 99.72%), lapatinib (#S2111, purity: 99.93%), gefitinib (#S1025, purity: 99.97%), PF-4708671 (#S2163, purity: 99.87%), LY2584702 (#S7698, purity: 98.15%), and RAD-001 (#S1120, purity: 99.77%) were obtained from Selleckchem (Houston, Texas, USA).

Techniques: MTT Assay, Control

Journal: American Journal of Cancer Research

Article Title: Duloxetine enhances the sensitivity of non-small cell lung cancer cells to EGFR inhibitors by REDD1-induced mTORC1/S6K1 suppression

doi: 10.62347/WMQV6643

Figure Lengend Snippet: Duloxetine and/or lapatinib reduces 70-kDa ribosomal protein S6 kinase 1 (p70S6K1)/mechanistic target of rapamycin complex 1 (mTORC1) activity. A. H1299 cells were treated with the indicated concentrations of lapatinib for 18 h. B. H1299 cells were treated with the indicated concentrations of duloxetine for 18 h. C. H1299 cells were treated with the indicated concentrations of duloxetine, milnacipran, venlafaxine and des venlafaxine for 18 h. D. H1299 cells were treated with 5 μM lapatinib and/or 10 μM duloxetine for 18 h. A-D. The indicated protein levels were estimated by western blot analysis. Data are representative of three independent experiments. DLX: duloxetine, LPT: lapatinib.

Article Snippet: Milnacipran (#S3140, purity: 99.99%), venlafaxine (#S1441, purity: 99.98%), desvenlafaxine (#S4113, purity: 99.72%), lapatinib (#S2111, purity: 99.93%), gefitinib (#S1025, purity: 99.97%), PF-4708671 (#S2163, purity: 99.87%), LY2584702 (#S7698, purity: 98.15%), and RAD-001 (#S1120, purity: 99.77%) were obtained from Selleckchem (Houston, Texas, USA).

Techniques: Activity Assay, Western Blot

Journal: American Journal of Cancer Research

Article Title: Duloxetine enhances the sensitivity of non-small cell lung cancer cells to EGFR inhibitors by REDD1-induced mTORC1/S6K1 suppression

doi: 10.62347/WMQV6643

Figure Lengend Snippet: Duloxetine, a serotonin-norepinephrine reuptake inhibitor, sensitizes EGFR inhibitors in the EGFR-TKI-resistant lung cancer cell line H1975 (L858R/T790M-mutant EGFR). A. H1975 cells were treated with 5 μM lapatinib, 5 μM gefitinib or 5 μM erlotinib with duloxetine for 48 h. B, C. H1975 cells were treated with lapatinib and duloxetine, milnacipran, venlafaxine or desvenlafaxine for 48 h. A, B. Cell viability was measured by MTT assay. The data are presented as the mean percentage of control ± SD (n=3). C. Cell death was detected as the percentage of Annexin V and/or PI positive cells. The data are presented as the mean percentage of control ± SD (n=3; *P<0.05, ***P<0.001, ns: not significantly different). DLX: duloxetine, ERL: erlotinib, GEF: gefitinib, LPT: lapatinib, PI: Propidium Iodide.

Article Snippet: Milnacipran (#S3140, purity: 99.99%), venlafaxine (#S1441, purity: 99.98%), desvenlafaxine (#S4113, purity: 99.72%), lapatinib (#S2111, purity: 99.93%), gefitinib (#S1025, purity: 99.97%), PF-4708671 (#S2163, purity: 99.87%), LY2584702 (#S7698, purity: 98.15%), and RAD-001 (#S1120, purity: 99.77%) were obtained from Selleckchem (Houston, Texas, USA).

Techniques: Mutagenesis, MTT Assay, Control

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: The Antinociceptive Effect of Milnacipran in the Monosodium Iodoacetate Model of Osteoarthritis Pain and Its Relation to Changes in Descending Inhibition

doi: 10.1124/jpet.112.199489

Figure Lengend Snippet: Baseline mechanical hypersensitivity prior to the injection of either milnacipran or saline (vehicle) in both phases of the MIA model Data presented as the median number of hind-limb withdrawals from 10 applications to each paw of 1-, 6-, and 8-g vF filaments. n = 8 in each group, in both phases of the model.

Article Snippet: Milnacipran (Tocris, Bristol, UK) was dissolved in physiologic saline.

Techniques: Injection, Saline

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: The Antinociceptive Effect of Milnacipran in the Monosodium Iodoacetate Model of Osteoarthritis Pain and Its Relation to Changes in Descending Inhibition

doi: 10.1124/jpet.112.199489

Figure Lengend Snippet: Mechanical hypersensitivity of the ipsilateral hind-paw is attenuated by milnacipran in both the early (A and B) and late phases (C and D) of the MIA model. (A and C) Injection of milnacipran (10 mg/kg i.p.) caused a significant reduction in mechanical hypersensitivity compared with predrug values (#P < 0.05; ##P < 0.01; Friedman test and Dunn’s post hoc comparisons) and with the effects of saline (*P < 0.05; **P < 0.01; Mann–Whitney U tests with the Bonferroni correction) in the early (A) (n = 8) and late (C) (n = 8) phases of the model. Data are displayed as median difference scores, with postinjection scores normalized to the predrug baseline. (B and D) AUCs were significantly larger after milnacipran than after saline (**P < 0.01; ***P < 0.001; Mann–Whitney U test) in both the early (B) and late (D) phases of the MIA model. MIL, milnacipran; SAL, saline.

Article Snippet: Milnacipran (Tocris, Bristol, UK) was dissolved in physiologic saline.

Techniques: Injection, Saline, MANN-WHITNEY

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: The Antinociceptive Effect of Milnacipran in the Monosodium Iodoacetate Model of Osteoarthritis Pain and Its Relation to Changes in Descending Inhibition

doi: 10.1124/jpet.112.199489

Figure Lengend Snippet: Milnacipran inhibits electrically evoked responses from deep dorsal horn neurons in the early (n = 14) and late (n = 21) phases of the MIA model and in naïve animals (n = 24). In all three animal groups, milnacipran significantly reduced C-fiber–evoked responses (C) and postdischarge (D), although Aβ–evoked responses (A) and Aδ-fiber–evoked responses (B) were inhibited only in naïve animals (versus predrug baseline; ***P < 0.001; paired t test or Wilcoxon matched-pairs signed rank test; below graphs). Effects were consistently greater in naïve animals compared with the late phase of the model (naïve versus late phase MIA; **P < 0.01; Kruskal–Wallis tests with Dunn’s post hoc comparisons or one-way ANOVAs and Bonferroni post hoc tests; above graphs). Data are displayed as the mean % inhibition from predrug values in box and whisker plots apposing the level of inhibition caused by a dose of milnacipran (10 mg/kg s.c.) in the three animal groups. Plus symbols represent means, bars represent the median, and error bars mark the range of data.

Article Snippet: Milnacipran (Tocris, Bristol, UK) was dissolved in physiologic saline.

Techniques: Inhibition, Whisker Assay

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: The Antinociceptive Effect of Milnacipran in the Monosodium Iodoacetate Model of Osteoarthritis Pain and Its Relation to Changes in Descending Inhibition

doi: 10.1124/jpet.112.199489

Figure Lengend Snippet: Blocking spinal α2-adrenoceptors reverses the inhibitory effects of milnacipran in naive animals (A and B) and in the early phase (C and D) but not in the late phase (E and F) of the MIA model (n = 7 per group). Milnacipran (s.c., 10 mg/kg) reduced responses from deep dorsal horn neurons to noxious mechanical and thermal stimuli in all three groups. In naïve animals (A and B) and in the early phase of the model (C and D), responses were reversed to predrug baseline values when the α2-adrenoceptor antagonist atipamezole (10 µg/50 µl) was administered (spinal). Atipamezole had no similar effect in the late phase of the model (E and F). Data are displayed as mean number of APs (± S.E.M.) evoked, plotted as a function of intensity of mechanical (A, C, and E) or thermal stimuli (B, D, and F) applied to the IL hind-paw. Data were analyzed with two-way RM ANOVAs and Bonferroni post hoc comparisons. Predrug versus +MIL: *P < 0.05; **P < 0.01; ***P < 0.001. +MIL versus +MIL+ATI: ^^^P < 0.001. Predrug versus +MIL+ATI: #P < 0.05; ###P < 0.001. MIL, milnacipran; ATI, atipamezole.

Article Snippet: Milnacipran (Tocris, Bristol, UK) was dissolved in physiologic saline.

Techniques: Blocking Assay

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: The Antinociceptive Effect of Milnacipran in the Monosodium Iodoacetate Model of Osteoarthritis Pain and Its Relation to Changes in Descending Inhibition

doi: 10.1124/jpet.112.199489

Figure Lengend Snippet: Blocking spinal 5-HT7 receptors in the late phase of the MIA model (n = 9) reversed the inhibitory effect of milnacipran on responses from deep dorsal horn neurons to noxious thermal stimuli. Milnacipran (10 mg/kg s.c.) had attenuated responses to high-intensity mechanical (A) and thermal stimuli (B), but responses to the latter were fully reversed to baseline values after spinal administration of the 5-HT7 receptor antagonist SB-269970 (1 µg/50 µl). In contrast, SB-269970 had only a partial reversal effect on the inhibition of mechanical responses. Data are plotted as the mean number of APs (± S.E.M) as a function of stimulus intensity, two-way RM ANOVAs with Bonferroni post hoc tests. The inhibitory effects of milnacipran on C-fiber–evoked responses (C) and postdischarge (D) were also fully reversed after SB-269970 administration. Data are plotted as the mean % of predrug values and displayed as box and whisker plots, with plus symbols representing means, bars representing the median, and error bars marking the range of data (analyzed with one-way RM ANOVAs and Bonferroni post hoc tests). Predrug versus +MIL: **P < 0.01; ***P < 0.001. Predrug versus MIL +SB-269970: ^^^P < 0.001. +MIL +SB-269970 versus +MIL: #P < 0.05; ###P < 0.001. MIL, milnacipran.

Article Snippet: Milnacipran (Tocris, Bristol, UK) was dissolved in physiologic saline.

Techniques: Blocking Assay, Inhibition, Whisker Assay

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: The Antinociceptive Effect of Milnacipran in the Monosodium Iodoacetate Model of Osteoarthritis Pain and Its Relation to Changes in Descending Inhibition

doi: 10.1124/jpet.112.199489

Figure Lengend Snippet: Spinal administration of the opioid receptor antagonist naloxone partially reverses the inhibitory effect of milnacipran in the early and late phases of the MIA model and in naïve animals (n = 9 per group). Milnacipran (10 mg/kg s.c.) reduced responses to noxious mechanical and thermal stimuli in all three animal groups and spinal application of the opioid receptor antagonist naloxone (50 µg/50 µl) significantly increased responses. In all groups, however, this reversal was only partial, with responses from deep dorsal horn neurons still significantly lower than predrug values. Data plotted as the mean number of APs (± S.E.M.) as a function of stimulus intensity. Analysis with two-way RM ANOVAs and Bonferroni post hoc tests. Predrug versus +MIL: ***P < 0.001. Predrug versus +MIL+NALOX: ^^P < 0.01; ^^^P < 0.001. +MIL versus +MIL+NALOX: #P < 0.05; ##P < 0.01; ###P < 0.001. MIL, milnacipran; NALOX, naloxone.

Article Snippet: Milnacipran (Tocris, Bristol, UK) was dissolved in physiologic saline.

Techniques: