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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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Related in: MedlinePlus

LND induces cellular ROS release and reduces cell viability.A and B, ROS in DB-1 cells was measured by 2′,7′-dichlorofluorescein (DCF) fluorescence. Shown are the representative flow cytometry histograms of samples treated with DMSO or the indicated drugs. C, mean fluorescence intensities of cells treated with DMSO or the indicated drugs. D and E, PI exclusion method was used to examine the viability of DB-1 cells treated with DMSO or indicated drugs. Shown are the percentages of PI-positive cells. The data presented are means of three samples. Error bars represent S.D. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (Student's t test). F, representative flow cytometry histograms for PI-stained cells from D. Percentages of PI-positive cells are indicated. Cells were treated with the indicated drugs for 48 (D and F) or 24 h (E). The following drug concentrations were used: LND, 150 μm; TTFA, 150 μm unless indicated otherwise; 3-NPA, 500 μm; N-acetylcysteine (NAC), 10 mm.
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Figure 4: LND induces cellular ROS release and reduces cell viability.A and B, ROS in DB-1 cells was measured by 2′,7′-dichlorofluorescein (DCF) fluorescence. Shown are the representative flow cytometry histograms of samples treated with DMSO or the indicated drugs. C, mean fluorescence intensities of cells treated with DMSO or the indicated drugs. D and E, PI exclusion method was used to examine the viability of DB-1 cells treated with DMSO or indicated drugs. Shown are the percentages of PI-positive cells. The data presented are means of three samples. Error bars represent S.D. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (Student's t test). F, representative flow cytometry histograms for PI-stained cells from D. Percentages of PI-positive cells are indicated. Cells were treated with the indicated drugs for 48 (D and F) or 24 h (E). The following drug concentrations were used: LND, 150 μm; TTFA, 150 μm unless indicated otherwise; 3-NPA, 500 μm; N-acetylcysteine (NAC), 10 mm.

Mentions: As part of the ETC, complex II is also a source of ROS. It mainly produces ROS from either the reduced FAD or ubiquinone site when downstream components of the ETC are blocked (40, 41). To examine whether LND induces intracellular ROS generation, we quantified the level of ROS by 2′,7′-dichlorofluorescein fluorescence (Fig. 4, A–C). Surprisingly, the level of ROS formed in LND-treated cells was even higher than the level in cells treated with TTFA (Fig. 4, A and C).


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 induces cellular ROS release and reduces cell viability.A and B, ROS in DB-1 cells was measured by 2′,7′-dichlorofluorescein (DCF) fluorescence. Shown are the representative flow cytometry histograms of samples treated with DMSO or the indicated drugs. C, mean fluorescence intensities of cells treated with DMSO or the indicated drugs. D and E, PI exclusion method was used to examine the viability of DB-1 cells treated with DMSO or indicated drugs. Shown are the percentages of PI-positive cells. The data presented are means of three samples. Error bars represent S.D. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (Student's t test). F, representative flow cytometry histograms for PI-stained cells from D. Percentages of PI-positive cells are indicated. Cells were treated with the indicated drugs for 48 (D and F) or 24 h (E). The following drug concentrations were used: LND, 150 μm; TTFA, 150 μm unless indicated otherwise; 3-NPA, 500 μm; N-acetylcysteine (NAC), 10 mm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4697178&req=5

Figure 4: LND induces cellular ROS release and reduces cell viability.A and B, ROS in DB-1 cells was measured by 2′,7′-dichlorofluorescein (DCF) fluorescence. Shown are the representative flow cytometry histograms of samples treated with DMSO or the indicated drugs. C, mean fluorescence intensities of cells treated with DMSO or the indicated drugs. D and E, PI exclusion method was used to examine the viability of DB-1 cells treated with DMSO or indicated drugs. Shown are the percentages of PI-positive cells. The data presented are means of three samples. Error bars represent S.D. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (Student's t test). F, representative flow cytometry histograms for PI-stained cells from D. Percentages of PI-positive cells are indicated. Cells were treated with the indicated drugs for 48 (D and F) or 24 h (E). The following drug concentrations were used: LND, 150 μm; TTFA, 150 μm unless indicated otherwise; 3-NPA, 500 μm; N-acetylcysteine (NAC), 10 mm.
Mentions: As part of the ETC, complex II is also a source of ROS. It mainly produces ROS from either the reduced FAD or ubiquinone site when downstream components of the ETC are blocked (40, 41). To examine whether LND induces intracellular ROS generation, we quantified the level of ROS by 2′,7′-dichlorofluorescein fluorescence (Fig. 4, A–C). Surprisingly, the level of ROS formed in LND-treated cells was even higher than the level in cells treated with TTFA (Fig. 4, A and C).

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