Resistance to arginine deiminase treatment in melanoma cells is associated with induced argininosuccinate synthetase expression involving c-Myc/HIF-1α/Sp4

Wen Bin Tsai, Isamu Aiba, Soo Yong Lee, Lynn G Feun, Niramol Savaraj, Macus Tien Kuo

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Abstract

Arginine deiminase (ADI)-based arginine depletion is a novel strategy under clinical trials for the treatment of malignant melanoma with promising results. The sensitivity of melanoma to ADI treatment is based on its auxotrophy for arginine due to a lack of argininosuccinate synthetase (AS) expression, the rate-limiting enzyme for the de novo biosynthesis of arginine. We show here that AS expression can be transcriptionally induced by ADI in melanoma cell lines A2058 and SK-MEL-2 but not in A375 cells, and this inducibility was correlated with resistance to ADI treatment. The proximal region of the AS promoter contains an E-box that is recognized by c-Myc and HIF-1α and a GC-box by Sp4. Through ChIP assays, we showed that under noninduced conditions, the E-box was bound by HIF-1α in all the three melanoma cell lines. Under arginine depletion conditions, HIF-1α was replaced by c-Myc in A2058 and SK-MEL-2 cells but not in A375 cells. Sp4 was constitutively bound to the GC-box regardless of arginine availability in all three cell lines. Overexpressing c-Myc by transfection upregulated AS expression in A2058 and SK-MEL-2 cells,whereas cotransfection with HIF-1α suppressed c-Myc-induced AS expression. These results suggest that regulation of AS expression involves interplay among positive transcriptional regulators c-Myc and Sp4, and negative regulator HIF-1α that confers resistance to ADI treatment in A2058 and SK-MEL-2 cells. Inability of AS induction in A375 cells under arginine depletion conditions was correlated by the failure of c-Myc to interact with the AS promoter.

Original languageEnglish
Pages (from-to)3223-3233
Number of pages11
JournalMolecular Cancer Therapeutics
Volume8
Issue number12
DOIs
StatePublished - Dec 1 2009

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Argininosuccinate Synthase
Melanoma
Arginine
Cell Line
arginine deiminase
Transfection
Clinical Trials

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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Resistance to arginine deiminase treatment in melanoma cells is associated with induced argininosuccinate synthetase expression involving c-Myc/HIF-1α/Sp4. / Tsai, Wen Bin; Aiba, Isamu; Lee, Soo Yong; Feun, Lynn G; Savaraj, Niramol; Kuo, Macus Tien.

In: Molecular Cancer Therapeutics, Vol. 8, No. 12, 01.12.2009, p. 3223-3233.

Research output: Contribution to journalArticle

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