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Characterization and inhibition of fatty acid synthase in pediatric tumor cell lines
Renee F Slade1, Dirk A Hunt, Mildred M Pochet
1Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA 30322, USA.
Abstract:
The current study characterizes the lipogenic enzyme fatty acid synthase (FAS; EC 2.3.1.85) in pediatric tumor cell lines of neural or neural crest origin [medulloblastoma (Daoy), malignant rhabdoid tumor of kidney (SM II), retinoblastoma (Y79), and neuroblastoma (SK-N-SH)]. Constitutive FAS content and activity in these lines were compared to human fibroblast cell line Hs27. Hs27 exhibits low levels of FAS and recapitulates enzyme status in normal human tissues under most physiological conditions. Western analysis detected significantly larger amounts of FAS protein in Y79 and SK-N-SH than Daoy, SM II and Hs27. Incorporation of radiolabeled malonyl-CoA into total cellular lipid revealed that enzyme activity correlated with amount. Increased FAS content and activity in Y79 and SK-N-SH relative to the other cell lines and Hs27, in particular, implied enzyme activation in retinoblastoma and neuroblastoma lineages. The enzyme also showed evidence of hormonal regulation, as dexamethasone induced FAS protein in Daoy and SK-N-SH. However, hormonal induction of FAS protein levels did not correlate with activity levels, which led us to speculate phosphorylation as a means of regulating the enzyme's activity. Finally, the FAS inhibitor cerulenin was investigated for its ability to suppress tumor cell growth. After four days of propagation, short-term treatment of cell lines with drug produced mean IC50s less than 10.5 micrograms/ml (i.e., 5.6 +/- 1.9 for SM II; 9.3 +/- 1.5 for Daoy; 10.2 +/- 0.2 for SK-N-SH; and 10.4 +/- 2.6 for Y79). Annexin V assays revealed that cerulenin initiated apoptosis. The antineoplastic properties of cerulenin documented here are consistent with prior studies showing its cytotoxic effects upon other types of cancer cells and illustrate the potential utility of FAS inhibition as a novel chemotherapeutic approach.
Insights
This study found that fatty acid synthase (FAS) is highly active in pediatric retinoblastoma and neuroblastoma cells. The FAS inhibitor cerulenin suppressed tumor growth and induced apoptosis, suggesting its potential as a cancer therapy.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Fatty acid synthase (FAS) is a key lipogenic enzyme involved in fatty acid synthesis.
- Dysregulation of FAS has been implicated in various cancers.
- Pediatric tumors of neural or neural crest origin present unique therapeutic challenges.
Purpose of the Study:
- To characterize fatty acid synthase (FAS) content and activity in pediatric tumor cell lines.
- To investigate the hormonal regulation of FAS in these cell lines.
- To evaluate the efficacy of the FAS inhibitor cerulenin as a potential chemotherapeutic agent.
Main Methods:
- Western blot analysis to quantify FAS protein levels.
- Radiolabeled malonyl-CoA incorporation assays to measure enzyme activity.
- Cell viability assays (IC50 determination) and Annexin V assays to assess cerulenin's effects.
Main Results:
- Significantly higher FAS protein levels and activity were observed in retinoblastoma (Y79) and neuroblastoma (SK-N-SH) cell lines compared to medulloblastoma (Daoy), malignant rhabdoid tumor (SM II), and fibroblast (Hs27) cells.
- Dexamethasone induced FAS protein levels in Daoy and SK-N-SH, but not activity, suggesting post-translational regulation like phosphorylation.
- Cerulenin demonstrated potent anti-proliferative effects with low IC50 values across all tested cell lines and induced apoptosis.
Conclusions:
- FAS is upregulated and potentially activated in specific pediatric neural crest-derived tumors.
- FAS activity may be regulated by mechanisms beyond protein expression, such as phosphorylation.
- Cerulenin exhibits significant antineoplastic properties against these pediatric tumor cell lines, highlighting FAS inhibition as a promising therapeutic strategy.
