Viability and oxidative response of human colorectal HCT-116 cancer cells treated with visfatin/eNampt in vitro.
Visfatin/eNampt is a novel adipokine, secreted by visceral and subcutaneous fat, which could be involved in the development of obesity-associated cancer. Only few studies revealed reactive oxygen species (ROS)-dependent action of visfatin in endothelial cells, myotubes and melanoma cells. The potential pro-apoptotic properties of visfatin/eNampt in human colorectal HCT-116 cells remain unknown. The aim of the study was to examine the effects of visfatin/eNampt on cell viability along with the determination of apoptosis/necrosis extent and ROS level in HCT-116 cells. Additionally antioxidant enzymes' activities (i.e catalase (CAT), gluthatione peroxidase (GSH-Px)), and lipid peroxidation intensity in HCT-116 cells line was evaluated. Viability of HCT-116 cells was decreased after visfatin/eNampt treatment for 24 hours. The number of apoptotic cells in tested cells treated with increasing visfatin/eNampt concentrations (10, 100, 250 ng/ml) was elevated compared to untreated cells (6.4%, 9.7%, 16% vs. 3.2%; respectively). After 24 hours in the visfatin/eNampt treated group (10 - 100 ng/ml) CAT and GSH-Px activities significantly increased and this observation was accompanied by the decrease of ROS level when compared to the control group. Interestingly ROS level (using DCF detection technique) and lipid peroxidation ratio were increased in cells stimulated by visfatin/eNampt in concentration of 250 ng/ml along with the decreased activity of selected antioxidant enzymes when compared to remaining study groups, including control. We concluded that visfatin/eNampt induces decrease of cell viability and apoptosis boost in human colorectal cancer HCT-116 cells line. Visfatin/eNampt affected the level of ROS as well as antioxidant capacity, however the association of ROS level and apoptosis rate was not linear. The role for visfatin/eNampt in cancer redox status in vitro may provide a greater insight into the association between fat derived visfatin/eNampt and its endocrine action in colorectal carcinoma cells.
Medical University of Silesia, School of Medicine with the Division of Dentistry, Department of Physiology in Zabrze, Zabrze, Poland. rbuldak@gmail.com.
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Buldak
Medical University of Silesia, School of Medicine with the Division of Dentistry, Department of Physiology in Zabrze, Zabrze, Poland.
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Gowarzewski
Medical University of Silesia, School of Medicine, Department of Internal Medicine and Clinical Pharmacology, Katowice, Poland.
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L
Buldak
Silesian University of Technology, Institute of Automatic Control, Biosystems Group, Department of Automatics, Electronics, and Informatics, Gliwice, Poland.
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Skonieczna
Medical University of Silesia, School of Medicine, Department of Gastroenterology and Hepatology, Katowice, Poland.
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M
Kukla
Medical University of Silesia, School of Public Health, Department of Human Nutrition, Katowice, Poland.
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R
Polaniak
Medical University of Silesia, School of Medicine with the Division of Dentistry, Department of Physiology in Zabrze, Zabrze, Poland.
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Zwirska-Korczala
D017209:Apoptosis / Q000187:drug effects; D002374:Catalase / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects*; D016207:Cytokines / Q000494:pharmacology*; D005979:Glutathione Peroxidase / Q000378:metabolism; D045325:HCT116 Cells; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D008315:Malondialdehyde / Q000378:metabolism; D009336:Necrosis; D054409:Nicotinamide Phosphoribosyltransferase / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism
D016207:Cytokines; D017382:Reactive Oxygen Species; D008315:Malondialdehyde; D002374:Catalase; D005979:Glutathione Peroxidase; D054409:Nicotinamide Phosphoribosyltransferase; C085455:nicotinamide phosphoribosyltransferase, human