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Induction of apoptosis and reactive oxygen species production by N-nitrosopiperidine and N-nitrosodibutylamine in human leukemia cells

✍ Scribed by Almudena García; Paloma Morales; Nuria Arranz; Eugenia Delgado; Joseph Rafter; Ana I. Haza

Publisher
John Wiley and Sons
Year
2008
Tongue
English
Weight
310 KB
Volume
28
Category
Article
ISSN
0260-437X

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✦ Synopsis

Abstract

N‐nitrosopiperidine (NPIP) and N‐nitrosodibutylamine (NDBA) belong to a group of N‐nitrosamines that are widely distributed in foodstuffs and the occupational environment. In the present study, the human promyelocytic leukemia cell line HL‐60, was used to characterize the apoptotic effects of N‐nitrosamines, and to examine the production of reactive oxygen species (ROS). Apoptotic cells were identified by (i) chromatin condensation (ii) flow cytometry analysis and (iii) poly(ADP‐ribose) polymerase (PARP) cleavage. NPIP and NDBA induced morphological changes consistent with apoptotic events in HL‐60 cells. Flow cytometry analysis showed that both N‐nitrosamines induced apoptotic cell death in a concentration and time dependent‐manner. It was observed that NDBA was stronger than NPIP, since it induced a significant apoptotic cell death after 18 h starting from a concentration of 2 mm, whereas NPIP was effective at 10 mm. Furthermore, PARP was markedly cleaved with 0.5 mm of NDBA and 5 mm of NPIP after treatments for 3 and 18 h, respectively. Finally, the ROS level was found to be elevated after 0.5 h of treatment with both N‐nitrosamines. Antioxidant N‐acetylcysteine (NAC) completely inhibited the ROS production induced by NPIP and NDBA. However, this action seems not to be associated with the apoptosis because NAC did not block N‐nitrosamines‐induced apoptosis. The data demonstrate that NPIP and NDBA induce apoptosis and ROS production in HL‐60 cells. Copyright © 2007 John Wiley & Sons, Ltd.

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