Nitric oxide levels in human preneoplastic colonocytes determine their susceptibility toward antineoplastic agents.
The efficacy of antineoplastic compounds can depend heavily on the genetic background of the cells exposed to the drugs. This becomes evident by the fact that HT-29 human colon cancer cells but not primary murine nontransformed colonocytes are efficiently submitted to apoptosis by the flavonoid flavone. By determining caspase-3 activation, plasma membrane disintegration, and nuclear fragmentation, we show here that flavone also does not promote apoptosis in preneoplastic NCOL-1 colonocytes derived from a nontransformed human biopsy specimen. In clear contrast, the antitumor drug camptothecin potently induces apoptosis in NCOL-1 cells associated with a specific down-regulation of the antiapoptotic factor bcl-XL at the mRNA and protein levels and with the activation of the mitochondrial apoptosis pathway. Confocal microscopy revealed an increased production of superoxide anion radicals in the mitochondria of NCOL-1 cells that preceded the apoptotic events. However, in the case of flavone, the mitochondrial oxygen radicals were effectively scavenged by physiological concentrations of nitric oxide (NO), whereas in the case of camptothecin, the available nitric oxide was rapidly scavenged by the production of large quantities of cytosolic superoxide anions. Increasing the levels of nitric oxide inside NCOL-1 cells by sodium nitroprusside prevented the apoptosis induction by camptothecin. Reducing the levels of nitric oxide by using the NO synthase inhibitor, Nomega-nitro-l-arginine methyl ester in NCOL-1 cells or using HT-29 cells that intrinsically have low NO levels enabled flavone to trigger the apoptosis pathway. In conclusion, our studies demonstrate that the intracellular levels of nitric oxide significantly change the apoptotic response to antineoplastic agents in colonic cells.
Wenzel
Uwe
U
Department of Food and Nutrition, Molecular Nutrition Unit, Hochfeldweg 2, d-85350 Freising-Weihenstephan, Germany. uwenzel@wzw.tum.de
Kuntz
Sabine
S
Daniel
Hannelore
H
D000970:Antineoplastic Agents / Q000494:pharmacology*; D002460:Cell Line; D045744:Cell Line, Tumor; D003106:Colon / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm; D019073:HT29 Cells; D006801:Humans; D009569:Nitric Oxide / Q000378:metabolism*; D011230:Precancerous Conditions / Q000209:etiology / Q000378:metabolism* / Q000473:pathology*
D000970:Antineoplastic Agents; D009569:Nitric Oxide