J Korean Endocr Soc.  2008 Aug;23(4):238-244.

Effects of Simvastatin on the Growth and Invasion of Anaplastic Thyroid Cancer Cells Lines

Affiliations
  • 1Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Korea.
  • 2Department of Internal Medicine, Asan Medical Center, University of Ulsan College of Medicine, Korea.

Abstract

BACKGROUND
Anaplastic thyroid carcinoma has grave prognosis with most patient dying within 6 months of diagnosis. 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors have been reported to have an anticancer effect in experimental and clinical studies. In this study, we investigated the effect of HMG-CoA reductase inhibitors on cell growth, invasiveness, adherence and signal transduction to evaluate the possibility of simvastatin as an agent for treatment of thyroid cancer. METHODS: The viability of simvastatin treated 3 thyroid cancer cell lines (FRO, WRO, and ARO) were determined. We evaluated the cell migration, anchorage-independent growth and invasion ability in anaplastic thyroid cell line. The expression and phosphorylation of focal adhesion kinase (FAK) and extracellular signal-regurated kinase (ERK) were determined by immunoblot analysis. RESULTS: Three thyroid cancer cell lines showed concentration dependent decrease of viability after treatment with 100~200 mM of simvastatin. Anaplastic ARO cell line showed the most predominant decrease in viability. In ARO cell lines, cell migration was decreased by concentration dependent manner after treatment with simvastatin (concentration > or = 5 mM). Anchorage independent colony formation also decreased after simvastatin (> or = 10 mM). Finally, immunoblot analysis revealed that the phosphorylation status of FAK and ERK decreased in time dependent manner following treatment with 10 mM of simvastatin. CONCLUSION: The results of this study suggest that simvstatin exerts a favorable effect on the progression and metastasis of thyroid cancer. However, further studies are needed to elucidate the related mechanisms and signal transductions prior to its therapeutic application.

Keyword

anaplastic thyroid cancer; chemotherapeutic agent; HMG-CoA reductase inhibitor

MeSH Terms

Acyl Coenzyme A
Cell Line
Cell Movement
Focal Adhesion Protein-Tyrosine Kinases
Humans
Hydroxymethylglutaryl-CoA Reductase Inhibitors
Neoplasm Metastasis
Oxidoreductases
Phosphorylation
Phosphotransferases
Prognosis
Signal Transduction
Simvastatin
Thyroid Gland
Thyroid Neoplasms
Acyl Coenzyme A
Focal Adhesion Protein-Tyrosine Kinases
Hydroxymethylglutaryl-CoA Reductase Inhibitors
Oxidoreductases
Phosphotransferases
Simvastatin
Thyroid Neoplasms

Figure

  • Fig. 1 Effect of simvastatin on thyroid cancer cells. Cells were exposed to the indicated concentration of simvastatin for 48 h, and cell viability was determined by CCK assay as described in "Materials and Methods". Data presented are mean ± SD for three independent experiments.

  • Fig. 2 Simvastatin inhibits migration and anchorage-independent colony formation of ARO cells. (A) Migration assay: The scratch wounds were created in a ARO cell monolayer and incubated with the indicated concentration of simvastatin. After 3 days, cells were photographed through inverted microscope under 100 X magnification. (B) Anchorage independent growth assay: ARO cells were cultured in soft agar plate with the indicated concentration of simvastatin as described in "Materials and Methods". After 11 days, cells were stained overnight in a solution of 1 ug/mL iodonitrophenyl tetrazolium (INT) and scanned by FluorS Multi-Imager (upper panel in B). The lower graph represents the percentage of colony density in upper panel in (B).

  • Fig. 3 Simvastatin inhibits invasiveness of ARO cells. ARO cells (2 × 105 cells) are seeded into the chamber in 300 uL culture medium and incubated with the indicated concentration of simvastatin in 24 well culture plate. (A) After 48 hours, invasive cells on lower surface of the ECMatrix gel were stained by dipping the insert in the staining solution and photographed through inverted microscope under 100 X magnification. (B) The stained cells were dissolved in 10% acetic acid, and colorimetric absorbance was measured at 560 nm. The graph represents the percentage of invasion value.

  • Fig. 4 Effect of simvastatin on activation of FAK and ERK in ARO cells. Cells were exposed to 10 uM simvastatin for 0~24h and activation patterns of FAK and ERK were determined by immunoblot analysis as described in methods. (A) Changes in activation of FAK and ERK. (B) Data quantified as densitometry units and presented as percentage of the control values. Data presented are mean ± SD for three independent experiments.


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