Article Figures & Data
Figures
- Fig. 1.
Effect of rapamycin on established lung tumors. A, experimental schema. Shaded area indicates rapamycin treatment. B, tumor incidence and multiplicity. C, tumor size. *, P = 0.0046. D, cellular proliferation, as assessed by Ki-67 staining in vehicle- and rapamycin-treated tumors. **, P = 0.003.
- Fig. 2.
Biomarker modulation after treatment with rapamycin given on a daily (5 of 7 d) regimen. A, representative images of phosphorylated S6 in NNK-induced lung tumors (TU) and large airways (LA) in the absence (VEH) or presence of rapamycin (RAPA). The relative intensity of staining for S6 phosphorylation in large airways and adenomas was averaged for five mice per group. *, P < 0.01; **, P = 0.005. B, immunoblotting of phosphorylated S6 (pS6) and total S6 in hepatic lysates. Each lane represents lysate from one mouse. C and D, activation of Akt and extracellular signal-regulated kinase (ERK) in NNK-induced lung tumors. Representative images of phosphorylation of Akt at S473 or T308 (C) or extracellular signal-regulated kinase (D) in lung tumors in the absence (VEH) or presence of rapamycin treatment (RAPA). The relative intensity of staining for Akt or extracellular signal-regulated kinase phosphorylation (pERK) in adenomas was averaged for five mice per group. ***, P = 0.05. Staining for Akt and extracellular signal-regulated kinase was predominantly cytoplasmic.
- Fig. 3.
Comparison of two dosing schedules of rapamycin. A, experimental schema. Mice were treated with rapamycin by i.p. injection 5 of 7 d (mice 4-9) or qod (mice 10-15). B, phosphorylation of S6 was assessed by immunohistochemistry in untreated controls (1-3), 4 h after treatment (4-6 and 10-12), or following the washout period of 72 h (7-9) or 48 h (13-15). Mean rapamycin blood levels (ng/mL) for each group (right). C, immunoblotting for levels of total S6 and phosphorylated S6 and 4E-binding protein 1 (p4E-BP1) in hepatic lysates. Mouse designation is as above.
- Fig. 4.
Continuous treatment with rapamycin prevents the development of NNK-induced tumors. A, experimental schema. B, tumor incidence and multiplicity. *, P < 0.0001. C, tumor size. **, P < 0.0001. D, phenotypic progression. E, cellular proliferation index (number of Ki-67 positive cells per tumor). ***, P < 0.0001.
- Fig. 5.
Biomarker modulation after treatment with rapamycin given on a qod regimen. A, representative images of phosphorylated S6 in lung tumors arising in the absence (VEH) or presence of rapamycin treatment (RAPA). The percentage of tumor cells with S6 phosphorylation intensity of ≥2 was quantified for tumors from five mice per group, *, P < 0.0001. B, immunoblotting of phosphorylated and total S6 in hepatic lysates. Each lane represents lysate from one mouse. C, activation of Akt. Representative images of phosphorylated Akt at S473 and T308 in NNK-induced lung tumors in the absence or presence of rapamycin. Staining for phosphorylation of Akt revealed cytoplasmic, homogeneous staining throughout the tumors. The relative intensity of staining in all lesions was averaged for five mice per group, and for S473 phosphorylation, staining intensity was analyzed in hyperplasic lesions and tumors. D, activation of extracellular signal-regulated kinase. Representative images of phosphorylated ERK in NNK-induced lung tumors in the absence or presence of rapamycin. Staining for extracellular signal-regulated kinase phosphorylation was predominantly cytoplasmic but was more intense near the periphery of tumors. Thus, a staining index was used to account for intensity as well as distribution of staining. E, tumor-infiltrating macrophages. Representative images of staining for the macrophage antigen F4-80 in normal lung (NL) and tumors (TU) arising in the absence (VEH) or presence of rapamycin treatment (RAPA). The number of macrophages was quantified for surrounding lung tissues and for all NNK-induced lesions.
Volume 13, Issue 7
