Cytochrome P-450-mediated O-demethylation: a route in the metabolic activation of etoposide (VP-16-213).

The antitumor agent VP-16-213 is oxidatively O-demethylated by rat liver microsomes and purified rat liver microsomal cytochrome P-450. 3-Methylcholanthrene can quantitatively induce O-demethylation of VP-16-213. The Km and Vmax values for O-demethylation by noninduced, phenobarbital-, and 3-methylcholanthrene-induced rat liver microsomes were found to be 130, 600, and 160 microM and 8.5, 11.8, and 15.6 nmol H2CO/min X mg protein, respectively. Mass spectrometric comparison of the product of O-demethylation of VP-16-213 with the synthetic metabolite resulted in identification of the orthodihydroxy derivative. In studies on the biological activity of the orthodihydroxy derivative, it was found to inactivate single- and double-stranded phiX174 DNA, to bind to calf thymus DNA and to be highly toxic against chinese hamster ovary cells.

COX-2 inhibitors act as radiosensitizer in tumor treatment.

Since cyclooxygenase-2 (COX-2) is overexpressed in malignant tissues, the COX-2 mediated signaling pathway has been recognized as potential target for therapeutic intervention. In most human tumors, COX-2 overexpression has been associated with tumor aggressiveness and poor clinical outcome. In vitro studies show inhibition of cell proliferation by selective COX-2 inhibitors alone, and enhancement of the response to irradiation. In vivo experimental reports demonstrate enhanced tumor response and impediment of tumor neovascularization following radiotherapy combined with COX-2 inhibition. Clinical studies on the combination of irradiation with COX-2 inhibitors are emerging. Taken together, the perspective for the combined approach of radiotherapy with COX-2 inhibition yields clinical significance since preclinical data demonstrate selective COX-2 inhibitors to act as radiosensitizer in tumor treatment.

Mutation spectrum in the lacI gene, induced by gamma-radiation in aqueous solution under oxic conditions.

Irradiation of DNA in a cellular environment leads to many types of DNA damage, resulting from various effects of gamma-radiation. One of these effects is the formation of water-derived radicals (e.g., .OH radicals), which are formed in the vicinity of DNA (indirect effect). To study the influence of the indirect effect on gamma-radiation-induced mutations, a newly constructed plasmid, containing the lacI gene as a target gene, was irradiated with 60Co gamma-radiation in aqueous solution, in the presence of oxygen. Under these circumstances, only .OH radicals will be responsible for the induced mutations. Sequence analysis of the gamma-radiation-induced mutations showed that 96% of all mutations were base pair substitutions, 87% of which occurred in the lacI gene, the others are formed in the lac operator part. All gamma-radiation-induced mutations in the lacI gene occurred exclusively on G:C base pairs, and no mutations at A:T base pairs could be detected. In the spontaneous mutation spectrum, 83% of all mutations were base pair substitutions, 35% of which occurred in the lacI gene and 48% in the lac operator part. Base pair substitutions on G:C base pairs were very similar in the gamma-radiation-induced and in the spontaneous mutation spectrum, implying a high contribution of .OH radicals to spontaneous mutagenesis. A:T to G:C transitions accounted for 10% of all spontaneous base pair substitutions in the lacI gene and are probably the result of effects, other than just .OH radicals. It can be concluded that .OH radicals are an important source for mutations at G:C base pairs. In this paper, the extracellular gamma-radiation-induced mutation spectrum is also compared to the previously obtained, intracellular gamma-radiation-induced mutation spectrum of the lacI gene. Comparison shows some differences, such as relative high amounts of mutations at A:T base pairs, G:C to T:A transversions and frameshift mutations in the intracellular gamma-radiation-induced mutation spectrum, as compared to the extracellular gamma-radiation-induced mutation spectrum. Since the extracellular gamma-radiation-induced mutation spectrum shows that .OH radicals are mainly responsible for base pair substitutions on G:C base pairs, mutations at A:T base pairs in the intracellular gamma-radiation-induced mutation spectrum are apparently the result of additional or other factors.