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Inhibition of Mitochondrial Complex II by the Anticancer Agent Lonidamine.

Guo L, Shestov AA, Worth AJ, Nath K, Nelson DS, Leeper DB, Glickson JD, Blair IA - J. Biol. Chem. (2015)

Bottom Line: However, the effect of LND on central energy metabolism has never been fully characterized.The ability of LND to promote cell death was potentiated by its suppression of the pentose phosphate pathway, which resulted in inhibition of NADPH and glutathione generation.Using stable isotope tracers in combination with isotopologue analysis, we showed that LND increased glutaminolysis but decreased reductive carboxylation of glutamine-derived α-ketoglutarate.

View Article: PubMed Central - PubMed

Affiliation: From the Penn Superfund Research and Training Program Center, Center of Excellence in Environmental Toxicology, and Department of Systems Pharmacology and Translational Therapeutics and.

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LND treatment alters the levels of TCA cycle metabolites.A and B, DB-1 cells were incubated with 150 μm LND or DMSO for 1 h. Levels of lactate (A) and metabolites in the TCA cycle (B) were measured by LC-MS. The levels of metabolites in LND-treated group were normalized with respect to the relevant metabolites in DMSO controls. C, a scheme of the TCA cycle. The changes observed in LND-treated cells are indicated. HepG2 (D), HCT116 (E), and HeLa (F) cells were incubated with DMSO or LND (150 μm) for 1 h. Levels of lactate and TCA metabolites were quantified by LC-MS. G and H, DB-1 cells were treated with DMSO (−), LND, 3-NPA, or TTFA at the indicated concentrations. Levels of succinate and lactate were quantified by LC-MS. For A, B, and D–H, the means of three samples are shown. Error bars represent S.D. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (Student's t test).
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Figure 1: LND treatment alters the levels of TCA cycle metabolites.A and B, DB-1 cells were incubated with 150 μm LND or DMSO for 1 h. Levels of lactate (A) and metabolites in the TCA cycle (B) were measured by LC-MS. The levels of metabolites in LND-treated group were normalized with respect to the relevant metabolites in DMSO controls. C, a scheme of the TCA cycle. The changes observed in LND-treated cells are indicated. HepG2 (D), HCT116 (E), and HeLa (F) cells were incubated with DMSO or LND (150 μm) for 1 h. Levels of lactate and TCA metabolites were quantified by LC-MS. G and H, DB-1 cells were treated with DMSO (−), LND, 3-NPA, or TTFA at the indicated concentrations. Levels of succinate and lactate were quantified by LC-MS. For A, B, and D–H, the means of three samples are shown. Error bars represent S.D. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (Student's t test).

Mentions: To examine the effect of LND on melanoma cell metabolism, we used LC-selected reaction monitoring/MS to analyze the metabolites in DB-1 melanoma cells treated with LND. As a monocarboxylate transporter inhibitor, LND is known to stimulate lactate accumulation in cells (25, 26). After 1 h of treatment with 150 μm LND, lactate was elevated nearly 5-fold over control cells (Fig. 1A). The levels of succinate and α-ketoglutarate in LND-treated DB-1 cells were found to increase, whereas the levels of citrate, fumarate, and malate decreased (Fig. 1, B and C). A 3–5-fold accumulation of succinate was also observed in a variety of cancer cell lines, including HepG2 (Fig. 1D), HCT116 (Fig. 1E), and HeLa (Fig. 1F).


Inhibition of Mitochondrial Complex II by the Anticancer Agent Lonidamine.

Guo L, Shestov AA, Worth AJ, Nath K, Nelson DS, Leeper DB, Glickson JD, Blair IA - J. Biol. Chem. (2015)

LND treatment alters the levels of TCA cycle metabolites.A and B, DB-1 cells were incubated with 150 μm LND or DMSO for 1 h. Levels of lactate (A) and metabolites in the TCA cycle (B) were measured by LC-MS. The levels of metabolites in LND-treated group were normalized with respect to the relevant metabolites in DMSO controls. C, a scheme of the TCA cycle. The changes observed in LND-treated cells are indicated. HepG2 (D), HCT116 (E), and HeLa (F) cells were incubated with DMSO or LND (150 μm) for 1 h. Levels of lactate and TCA metabolites were quantified by LC-MS. G and H, DB-1 cells were treated with DMSO (−), LND, 3-NPA, or TTFA at the indicated concentrations. Levels of succinate and lactate were quantified by LC-MS. For A, B, and D–H, the means of three samples are shown. Error bars represent S.D. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (Student's t test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 1: LND treatment alters the levels of TCA cycle metabolites.A and B, DB-1 cells were incubated with 150 μm LND or DMSO for 1 h. Levels of lactate (A) and metabolites in the TCA cycle (B) were measured by LC-MS. The levels of metabolites in LND-treated group were normalized with respect to the relevant metabolites in DMSO controls. C, a scheme of the TCA cycle. The changes observed in LND-treated cells are indicated. HepG2 (D), HCT116 (E), and HeLa (F) cells were incubated with DMSO or LND (150 μm) for 1 h. Levels of lactate and TCA metabolites were quantified by LC-MS. G and H, DB-1 cells were treated with DMSO (−), LND, 3-NPA, or TTFA at the indicated concentrations. Levels of succinate and lactate were quantified by LC-MS. For A, B, and D–H, the means of three samples are shown. Error bars represent S.D. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (Student's t test).
Mentions: To examine the effect of LND on melanoma cell metabolism, we used LC-selected reaction monitoring/MS to analyze the metabolites in DB-1 melanoma cells treated with LND. As a monocarboxylate transporter inhibitor, LND is known to stimulate lactate accumulation in cells (25, 26). After 1 h of treatment with 150 μm LND, lactate was elevated nearly 5-fold over control cells (Fig. 1A). The levels of succinate and α-ketoglutarate in LND-treated DB-1 cells were found to increase, whereas the levels of citrate, fumarate, and malate decreased (Fig. 1, B and C). A 3–5-fold accumulation of succinate was also observed in a variety of cancer cell lines, including HepG2 (Fig. 1D), HCT116 (Fig. 1E), and HeLa (Fig. 1F).

Bottom Line: However, the effect of LND on central energy metabolism has never been fully characterized.The ability of LND to promote cell death was potentiated by its suppression of the pentose phosphate pathway, which resulted in inhibition of NADPH and glutathione generation.Using stable isotope tracers in combination with isotopologue analysis, we showed that LND increased glutaminolysis but decreased reductive carboxylation of glutamine-derived α-ketoglutarate.

View Article: PubMed Central - PubMed

Affiliation: From the Penn Superfund Research and Training Program Center, Center of Excellence in Environmental Toxicology, and Department of Systems Pharmacology and Translational Therapeutics and.

Show MeSH
Related in: MedlinePlus