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. 2010 Jul;9(7):1968-76.
doi: 10.1158/1535-7163.MCT-10-0062. Epub 2010 Jun 29.

MEK inhibitor PD0325901 significantly reduces the growth of papillary thyroid carcinoma cells in vitro and in vivo (V体育官网入口)

Ying C Henderson  1 , Yunyun Chen, Mitchell J Frederick, Stephen Y Lai, Gary L Clayman
Affiliations

MEK inhibitor PD0325901 significantly reduces the growth of papillary thyroid carcinoma cells in vitro and in vivo

"V体育2025版" Ying C Henderson et al. Mol Cancer Ther. 2010 Jul.

Abstract

Papillary thyroid carcinomas (PTC) are the most common type of thyroid malignancy. Most PTC carry one of the two mutations, RET/PTC rearrangement or BRAF mutation. Both mutations are able to activate the mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK/ERK) signaling transduction pathway leading to cellular proliferation, differentiation, and apoptosis. PD0325901 is a specific MEK1/2 inhibitor and therefore is a promising drug to treat thyroid cancers with either RET/PTC or BRAF mutation. In this study we tested the effects of PD0325901 on PTC cells harboring either mutation in vitro by growth curves and Western blots and in vivo using a murine orthotopic xenograft model. We found that 50% growth inhibition (GI(50)) by PD0325901 was 11 nmol/L for the PTC cells with the RET/PTC1 rearrangement and 6. 3 nmol/L for PTC cells with a BRAF mutation, with both concentrations readily achievable in serum. After 1 week of oral administration of PD0325901 (20-25 mg/kg/day) in mice, no tumor growth was detected in mice inoculated with PTC cells bearing a BRAF mutation. For PTC with the RET/PTC1 rearrangement, the average tumor volume of the orthotopic tumor was reduced by 58% as compared with controls. In conclusion, our data suggested that PTC cells carrying a BRAF mutation were more sensitive to PD0325901 than were PTC cells carrying the RET/PTC1 rearrangement. Our findings support the clinical evaluation of PD0325901 for patients with PTC and potentially other carcinomas with BRAF mutations VSports手机版. .

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Conflict of interest statement

Disclosure of Potential Conflicts of Interest

No potential conflicts of interest were disclosed.

VSports注册入口 - Figures

Figure 1
Figure 1
PD0325901 inhibits PTC cell growth in vitro. A, PTC cells (K2 and TPC-1) were treated with PD0325901 for 2 days at 0.0064, 0.032, 0.16, 0.8, 4, 20, or 100 nmol/L. The MTT assay was used to determine the number of cells at each concentration by measuring the absorption at 570 nmol/L. The percent inhibition was calculated using Prism 3.0 software. The PD0325901 concentrations are shown in log scale (X scale). Each concentration was determined in triplicate for each type of PTC cell. This experiment was repeated twice. B, PTC cells (K2 and TPC-1) were treated with PD0325901 for 4 days at 1, 10, or 100 nmol/L. The MTT assay was used to determine the number of cells at each concentration by measuring the absorption at 570 nmol/L each day. Each concentration was determined in triplicates for each type of PTC cell. This experiment was repeated once.
Figure 2
Figure 2
PD0325901 suppresses the expression of p-ERK1/2 and induces apoptosis in PTC cells. A, K2 cells (top) or TPC-1 cells (bottom) were treated for 1 hour at 37°C with 1,000, 100, 10, 1, or 0.1 nmol/L of PD0325901. B, K2 (top) or TPC-1 (bottom) cells were treated with 100 nmol/L of PD0325901 once at 0 hour and cells were harvested at 1, 6, 8, 24, 48, 72, or 96 hours. Western blot analysis showed the expression of p-ERK1/2 and cleaved PARP (cPARP). Total ERK1/2 and actin were used as loading controls. Cells treated with DMSO (0 hour) were used as a positive control for the expression of p-ERK1/2, and cells treated with 1 μmol/L staurosporine (+) for 8 hours were used as a positive control for apoptosis. The Western blot analysis was repeated twice. C, K2 or TPC-1 cells were treated with 100 nmol/L PD0325901 for 4 days. The Cell Death Detection ELISAPLUS photometric enzyme immunoassay was used to detect DNA fragmentation with absorbance at 405 nm and subtracted from a background fluorescence at 490 nm. Each data point was determined in duplicate and the assay was repeated twice. Cells treated with DMSO (DMSO) and lysis buffer only (buffer) in ELISA were used as negative controls. Cells treated with 1 μmol/L staurosporine for 24 hours (+ control) were used as a positive control for apoptosis.
Figure 3
Figure 3
PD0325901 inhibits tumor growth in mice. A, in the control group, images from Xenogen showed 8 mice inoculated with PTC cells carrying a BRAF mutation before treatment (baseline, 6 days after inoculation, top) and images after treatment with vehicle for 5 days (2nd panel). In the treatment group, 7 mice inoculated with K2 cells before treatment (baseline, 6 days after inoculation, 3rd panel) and images after treatment with 25 mg/kg/day of PD0325901 (PD) for 5 days (bottom). B, tumor volume (mm3) was calculated after mice were sacrificed and the tumor size was measured by a caliper using the formula: length (mm) × width (mm) × depth (mm). Both mice treated with PD0325901 (PD) and vehicle are shown here. C, Kaplan-Meier survival curves are shown for mice inoculated with K2 (top) or TPC-1 (bottom) and treated with vehicle or PD0325901. The in vivo study was repeated twice. D, mice (each identified by its ear tag number) inoculated with K2 cells (top) or TPC-1 cells (bottom) were treated with 20 mg/kg PD0325901 for 1, 3, or 24 hours. Tumors were harvested at the indicated time points and protein extracts were prepared. The expression of p-ERK1/2 was detected by Western blot analysis. Total ERK1/2 was used as a loading control. Mice treated with vehicle were used as positive controls for the expression of p-ERK1/2. Each mouse is identified by its tag number.
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