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Apoptosis

, Volume 10, Issue 2, pp 359–368 | Cite as

α-lipoic acid induces apoptosis in human colon cancer cells by increasing mitochondrial respiration with a concomitant O2 −.-generation

  • U. WenzelEmail author
  • A. Nickel
  • H. Daniel
Article

Abstract

The antioxidant α-lipoic acid (ALA) has been shown to affect a variety of biological processes associated with oxidative stress including cancer. We determined in HT-29 human colon cancer cells whether ALA is able to affect apoptosis, as an important parameter disregulated in tumour development. Exposure of cells to ALA or its reduced form dihydrolipoic acid (DHLA) for 24 h dose dependently increased caspase-3-like activity and was associated with DNA-fragmentation. DHLA but not ALA was able to scavenge cytosolic O2 −. in HT-29 cells whereas both compounds increased O2 − .-generation inside mitochondria. Increased mitochondrial O2 − .-production was preceded by an increased influx of lactate or pyruvate into mitochondria and resulted in the down-regulation of the anti-apoptotic protein bcl-XL. Mitochondrial O2 −.-generation and apoptosis induced by ALA and DHLA could be prevented by the O2 − .-scavenger benzoquinone. Moreover, when the lactate/pyruvate transporter was inhibited by 5-nitro-2-(3-phenylpropylamino) benzoate, ALA- and DHLA-induced mitochondrial ROS-production and apoptosis were blocked. In contrast to HT-29 cells, no apoptosis was observed in non-transformed human colonocytes in response to ALA or DHLA addition. In conclusion, our study provides evidence that ALA and DHLA can effectively induce apoptosis in human colon cancer cells by a prooxidant mechanism that is initiated by an increased uptake of oxidizable substrates into mitochondria.

Keywords

HT-29 human colon cancer cells mitochondrial apoptosis pathway monocarboxylate transporter superoxide anion generation 

Abbreviations

Ac-DEVD-AMC

acetyl-aspartyl-glutamyl-valyl-aspartyl-amino-4-methyl-coumarine

ALA

Alpha-lipoic acid

CHAPS

3-[(cholamidopropyl)-dimethyl-ammonium]-1-propane-sulfonate

CLSM

Confocal laser scanning microscopy

DTT

dithiotreitol

GAP-DH

glyceraldehyd-3-phosphate dehydrogenase

ModEM

modified Eagle medium

NPPB

5-nitro-2-(3-phenylpropylamino)benzoate

PDH

pyruvate-dehydrogenase

proxyl fluorescamine

5-(2-carboxyphenyl)-5-hydroxy-1-((2,2,5,5-tetramethyl-1-oxy- pyrrolidin-3-yl)methyl)-3-phenyl-2-pyrrolin-4-one, potassium salt

ROS

reactice oxygen species

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Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  1. 1.Molecular Nutrition Unit, Department of Food and NutritionTechnical University of MunichFreisingFRG
  2. 2.Institute of Nutritional SciencesFreising-Weihenstephan

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