J Korean Soc Endocrinol.  2006 Apr;21(2):125-131. 10.3803/jkes.2008.21.2.125.

Role of 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside in the Growth Regulation of Anaplastic Thyroid Cancer Cells Lines

• Song JY
• Kim TY
• Kim WB
• Shong YK
• Rhee YS
• Suck JH
• Hong SJ

• KMID: 1511951
• DOI: http://doi.org/10.3803/jkes.2008.21.2.125

Abstract

BACKGROUND: Anaplastic thyroid carcinoma is one of the most aggressive human cancers with a median survival of only 6 months. Local surgical tumor debulking combined with radio-chemotherapy is generally used to treat this malady, but the low success rate has prompted the search for new therapeutic targets. We used 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) as an AMP-activated protein kinase (AMPK) activator to induce growth suppression and apoptosis in the anaplastic thyroid carcinoma cells.
METHODS
We investigated the effect of AICAR on the proliferation of thyroid cancer cell lines (ARO, WRO and FRO) by performing methyl-thiazoletetrazolium bromide assay. We wanted to see the effect of AICAR on the apoptosis and cell cycle of the thyroid cancer cells, and we wanted to determine the mechanism of these changes.
RESULTS
The proliferation of all thyroid cancer cell lines was significantly inhibited by administration of AICAR. FRO was the most susceptible cell line to AICAR treatment and so further studies were then performed with this cell line. The suppressive effect of AICAR on cell proliferation was related with phosphorylation of AMPK and the increased apoptosis. Also, cell cycle analysis revealed that progression to the G2-M phase was arrested (S-phase arrest) by AICAR treatment. S-phase arrest was associated with the increased protein expression of p21.
CONCLUSION
In the anaplastic thyroid cancer cell lines, AICAR inhibited proliferation due to the arrest in the S-phase; this was accompanied with the increased expression of p21. Overall, AMPK activation by AICAR or any other pharmacological agent could be a tempting potential target for thyroid cancer therapy.

MeSH Terms

Aminoimidazole Carboxamide
AMP-Activated Protein Kinases
Apoptosis
Cell Cycle
Cell Line
Cell Proliferation
Humans
Phosphorylation
Thyroid Gland*
Thyroid Neoplasms*
AMP-Activated Protein Kinases
Aminoimidazole Carboxamide

Figure

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  Fig. 1 Effects of 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR) on the proliferation of ARO, FRO, and WRO cells. Each cell was treated by various concentration of 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) for 72 hours. The cell survival was evaluated by methyl-thiazoletetrazolium bromide assay.

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  Fig. 2 Effects of 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR) on the proliferation of FRO cells. FRO cells were treated by 200 µM of AICAR for 0, 24, 48, 72, and 96 hours. The cell survival was evaluated by methyl-thiazoletetrazolium bromide assay.

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  Fig. 3 Effects of 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) on apoptosis of FRO cells. Apoptosis was measured by cell death detection ELISA plus kit (Roche applied science). FRO cells were treated by 0, 200, and 400 µM of AICAR for 0, 2, 6, and 12 hours, respectively.

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  Fig. 4 Effects of 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) on cell cycle of FRO cells. FRO cells were treated by 0, 200, and 400 µM of AICAR for 0, 2, 6, and 12 hours, respectively. Cell cycle was measured by propodium iodide staining and flow cytometric analysis.

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  Fig. 5 Effects of 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) on protein expression of phophorylated AMPK (pAMPK), AMPK, and p21. FRO cells were treated by 200 µM of AICAR for 0, 1, 6, 12, 24, and 48 hours, respectively.

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  Fig. 6 Effects of 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR) on protein expression of phophorylated AMPK (pAMPK), AMPK, and p21 FRO cells were treated by 0, 100, 200, and 400 µM of AICAR for 48 hours.

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