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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Repurposing the serotonin agonist Tegaserod as an anticancer agent in melanoma: molecular mechanisms and clinical implications

Fig. 4

Tegaserod (TM) delays tumor growth, induces tumor cell apoptosis and inhibits Akt and p-S6 phosphorylation in vivo. a C57BL/6 J mice were subcutaneously injected with 5 × 105 B16F10 cells. Seven days post-tumor injection, mice were randomized and into two groups and treated daily with 5 mg/kg of Tegaserod or vehicle for five consecutive days. a Tumor volume was measured for 18 days after which mice were sacrificed (n = 6–8). b Mice were sacrificed on Day 13 post tumor-inoculation and immunohistochemical staining of tumor tissue for p-S6 is shown (a representative image of n = 6 mice is shown). A third of pictures from tumors of mice treated with vehicle were classified as ‘high positive’ for p-S6 compared to only 1 slide from TM treated mice (3–5 pictures from different fields of view were obtained of tumors from each independent mouse, for a total of 26 and 18 tumor pictures for vehicle and TM treated mice respectively). c, left panel Immunoblots of tumor lysates from TM or control treated mice confirmed decreased Akt (Ser473) and S6 (Ser235/6) phosphorylation (n = 6–9 mice, with 3 mice being shown on one immunoblot) quantified in c, right panel. d Mice were sacrificed on Day 13 post tumor-inoculation and d, left panel immunohistochemical staining of tumor tissue revealed that tumors of mice treated with TM had increased active Caspase-3 expression (a representative image of n = 6 is shown). d, right panel. The relative score distribution of tumor slides is shown (3 pictures from different fields of view were obtained of tumors from each independent mouse (n = 6), for a total of 18 tumor pictures for each stain and treatment group. e C57BL/6 J mice were intravenously injected with 2 × 105 B16F10 cells. Starting at day 1 post inoculation, mice were treated with 5 mg/kg of Tegaserod or vehicle three times a week. Mice were sacrificed at Day 14 post tumor inoculation and lung metastases counted with representative lung images shown in the right panel (n = 10). f C57BL/6 J mice were intravenously injected with 105 B16F10 cells. Starting at day 1 post inoculation, mice were treated with 5 mg/kg of Tegaserod or vehicle three times a week till day 17 post-inoculation. Mice were monitored for survival (n = 6-7). All Scale bars indicate 50 μm. Error bars in the all experiments indicate SEM. *P < 0.05 as determined by a determined by a Student’s t-test (unpaired, 2 tailed) or and log-rank test for analysis of Kaplan Meier survival curves

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