NVP-AUY922, a novel HSP90 inhibitor, inhibits the progression of malignant pheochromocytoma in vitro and in vivo

Figures & data

Figure 1 17-AAG and NVP-AUY922 inhibit the proliferation of PC12 cells.

Notes: PC12 cells were treated with varying concentrations of 17-AAG or NVP-AUY922 for 24, 48, or 72 h. (A) HSP70 protein levels after exposure to 17-AAG or NVP-AUY922. All protein levels are expressed relative to the actin loading CTR and are normalized to the levels of the CTR. (B) Time-dependent and (C) dose-dependent loss of PC12 cell viability in response to NVP-AUY922. *P<0.05.
Abbreviations: 17-AAG, 17-allylamino-17-demethoxygeldanamcyin; HSP70, heat-shock protein 70; CTR, control.

Figure 2 NVP-AUY922 inhibits PC12 cell migration.

Notes: (A) Mean number of migrated cells per microscopic field was determined in triplicate. *P<0.05. Representative images are shown of (B) PC12 cells incubated with serum-free medium, (C) PC12 cells incubated with 75 nM NVP-AUY922 in serum-free medium, and (D) PC12 cells incubated with 150 nM NVP-AUY922 in serum-free medium. Magnification ×20.
Abbreviation: CTR, control.

Figure 3 Induction of cell-cycle arrest by NVP-AUY922 in PC12 cells.

Notes: (A) Cell-cycle progression was evaluated by PI staining and flow cytometry after treatment with 0, 100, 200, or 400 nM NVP-AUY922. (B) Percentage of PC12 cells in G1, G2, or S phase after treatment with NVP-AUY922. (C) Expression levels of cyclin B1 after exposure to NVP-AUY922 for 24 h. *P<0.05 and **P<0.01.
Abbreviations: PI, propidium iodide; CTR, control.

Figure 4 NVP-AUY922 induces apoptosis in PC12 cells.

Notes: (A) Detection of apoptosis using Annexin V and PI staining in PC12 cells after exposure to NVP-AUY922. (B) Percentage of PC12 cells undergoing apoptosis after exposure to NVP-AUY922. (C) Expression levels of Bax after exposure to NVP-AUY922 for 48 h. *P<0.05 and **P<0.01.
Abbreviations: PI, propidium iodide; CTR, control; PE-A, phycoerythrin; FITC-A, protein A fluorescein isothiocyanate.

Figure 5 NVP-AUY922 inhibits the growth of PCC tumors.

Notes: (A) Tumor volume is shown in cubic milimeters, which is calculated as 0.5236× (length × width × height) in mice treated with either vehicle or NVP-AUY922 (35 mg/kg) for 15 days. (B) Representative images of tumor appearance after treatment with vehicle or NVP-AUY922. (C) Mean tumor weights of NVP-AUY922- and vehicle-treated mice. (D) Mean body weights of NVP-AUY922- and vehicle-treated mice. (E) Percentage of apoptotic cells in PC12 xenografts of NVP-AUY922- and vehicle-treated mice. *P<0.05 and **P<0.01.
Abbreviations: PCC, pheochromocytoma; CTR, control.

Figure 6 NVP-AUY922 modulates MEK/ERK and PI3K/AKT signaling in PC12 cells.

Notes: (A) Effects of NVP-AUY922 on the expression levels and phosphorylation of HSP70, AKT, MEK, and ERK. (B) Densitometry was performed for the Western blots shown in (A).
Abbreviations: ERK, extracellular signal-regulated kinase; HSP70, heat-shock protein 70; CTR, control; p-AKT, phosphorylated AKT; p-ERK, phosphorylated ERK; p-MEK, phosphorylated MEK.