J Korean Endocr Soc.  2010 Mar;25(1):28-36. 10.3803/jkes.2010.25.1.28.

Effects of alpha-lipoic Acid on Differentiation of Thyroid Cancer Cells

• Kim WG
• Han DH
• Choi HJ
• Kim EY
• Kim TY
• Shong YK
• Kim WB

• KMID: 2177858
• DOI: http://doi.org/10.3803/jkes.2010.25.1.28

Abstract

BACKGROUND
Induction of re-differentiation is necessary for the proper treatment of patients with recurrent or metastatic differentiated thyroid cancer (DTC) because cancer cells show de-differentiation in about 30% of these patients. In this study, we evaluated the expression of thyroid specific genes after treatment with various agents to induce re-differentiation in the follicular thyroid cancer cell line FTC-133.
METHODS
FTC-133 cells were treated with U0126, LY294002, trichostatin A, retinoic acid (RA), 5'-azacytidine and alpha-lipoic acid (ALA). We evaluated mRNA expression of thyroid specific genes, thyroglobulin (Tg), sodium iodine symporter (NIS), PAX-8 and TTF-1 by reverse transcriptase polymerase chain reaction (PCR). Quantified expression of Tg mRNA was also evaluated by real-time PCR.
RESULTS
The expression of Tg mRNA increased after 48 h of treatment with 0.1 uM RA and the expression of Tg mRNA and TTF-1 mRNA increased after 48-72 h of treatment with ALA (10~100 uM). There was no change in thyroid specific gene expression by the other agents. Increased expression of Tg mRNA was confirmed by real-time PCR (1.3 times by 10 uM ALA and 3.6 times by 100 uM ALA). There was no basal NIS mRNA expression in FTC-133 cells and none of the tested agents induced expression of NIS mRNA. There was no change in phosphorylation of AMPK1-alpha after ALA treatment of FTC-133 cells.
CONCLUSION
ALA increases mRNA expression of Tg and TTF-1 of FTC-133 thyroid cancer cells and these effects are not mediated by activation of AMP kinase. The finding that ALA could be a potential re-differentiation inducing agent in thyroid cancer cells is novel. Further studies are needed to elucidate the mechanism of induction of re-differentiation. Furthermore, the effect of ALA on NIS expression and on iodine uptake should be evaluated using diverse thyroid cancer cell lines.

Keyword

Cancer; Differentiation; Lipoic acid; Thyroglobulin; Thyroid

MeSH Terms

Adenylate Kinase
Butadienes
Cell Line
Chromones
Gene Expression
Humans
Hydroxamic Acids
Iodine
Ion Transport
Morpholines
Nitriles
Phosphorylation
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger
Sodium
Thioctic Acid
Thyroglobulin
Thyroid Gland
Thyroid Neoplasms
Tretinoin
Adenylate Kinase
Butadienes
Chromones
Hydroxamic Acids
Iodine
Morpholines
Nitriles
RNA, Messenger
Sodium
Thioctic Acid
Thyroglobulin
Tretinoin

Figure

• 

  Fig. 1. Cell survival curves measured by cell counting kit-8 after 48 hours of treatment with re-differentiation agents in FTC-133 cells. U0126; selective inhibitor of MEK1/2, LY294002; selective inhibitor of PI3K, TSA; trichostatin A. U0126 (A), LY294002 (B), TSA (C), Retinoic acid (D), 5-Azacytidine (E), a-Lipoic acid (F).

• 

  Fig. 2. Results of reverse transcriptase polymerase chain reaction after 48 hours of treatment with re-differentiation agents in FTC-133 cells. GAPDH, glyceraldehydes 3-phosphate dehydrogenase; Tg, thyroglobulin; PAX-8, paired box 8; TTF-1, thyroid transcription factor-1; NIS, sodium iodine symporter; (+), positive control (human thyroid tissue); (−), negative control (no template); C, control without vehicle; U0126, selective inhibitor of MEK1/2; LY294002, selective inhibitor of PI3K; TSA, trichostatin A; RA, retinoic acid; AZT, 5’-azacytidine. U0126 (A), LY294002 (B), TSA (C), RA (D), AZT (E).

• 

  Fig. 3. Results of reverse transcriptase polymerase chain reaction after 72 hours of treatment with α-lipoic acids in FTC-133 cells. GAPDH, glyceraldehydes 3-phosphate dehydrogenase; Tg, thyroglobulin; PAX-8, paired box 8; TTF-1, thyroid transcription factor-1; NIS, sodium iodine symporter; (+), positive control (human thyroid tissue); (−), negative control (no template); C, control without vehicle.

• 

  Fig. 4. Results of real-time polymerase chain reaction after 72 hours of treatment with α-lipoic acids in FTC-133 cells.

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  Fig. 5. Western blotting of phospho-5-AMP-activated protein kinase (AMPK)-1α and total-AMPK1-α protein after treatment of α-lipoic acids during 30∼60 minutes (A) and 24–48 hours (B) in FTC-133 cells.

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