Apoptosis Induced by N-Nitrosamines in Two Cancer Cell Lines英文文献资料.docVIP

Journal of Environmental Protection, 2010, 1, 314-323 doi:10.4236/jep.2010.13037 Published Online September 2010 (http://www.SciRP.org/journal/jep) Apoptosis Induced by N-Nitrosamines in Two Cancer Cell Lines Paloma Morales, Nuria Arranz, Ana I. Haza Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, Madrid, Spain. Email: hanais@vet.ucm.es Received May 20 , 2010; revised July 1 , 2010; accepted July 3 , 2010. th st rd ABSTRACT In the present study, we investigated the induction of apoptosis by N-nitrosopyrrolidine (NPYR) and N-nitrosodimethy- lamine (NDMA) in two human cell lines: HL-60 (leukemia) and HepG2 (hepatoma). Apoptotic cells were identified by: 1) chromatin condensation, 2) flow cytometry analysis and 3) poly (ADP-ribose) polymerase cleavage. Both cell lines exhibited morphological changes consistent with apoptotic events following treatment with N-nitrosamines. Flow cy- tometry analysis showed that both N-nitrosamines induced apoptotic cell death in a concentration and time dependent- manner. NPYR was stronger than NDMA, since it induced a significant apoptotic cell death after 72 h starting from a concentration of 10 mM, whereas NDMA was effective at 27 mM. Furthermore, NPYR and NDMA caused the cleavage of PARP in HL-60 cells whereas no PARP cleavage was detected in HepG2 cells. However, NPYR- and NDMA- induced cell death in HepG2 cells was prevented by specific caspase inhibitors. Caspase-8 mediated main pathway and was responsible for 76% (NPYR) and 64% (NDMA) inhibition of apoptosis. The data demonstrate that NPYR and NDMA induce apoptosis in HL-60 and HepG2 cell lines via caspase-dependent pathway. Keywords: N-nitrosopyrrolidine, N-nitrosodimethylamine, Apoptosis, Caspases, HL-60 Cells, HepG2 Cells 1. Introduction acti