Metabolic detoxification: implications for thresholds.

Abstract

The fact that chemical carcinogenesis involves single, isolated, essentially irreversible molecular events as discrete steps, several of which must occur in a row to finally culminate in the development of a malignancy, rather suggests that an absolute threshold for chemical carcinogens may not exist. However, practical thresholds may exist due to saturable pathways involved in the metabolic processing, especially in the metabolic inactivation, of such compounds. An important example for such a pathway is the enzymatic hydrolysis of epoxides via epoxide hydrolases, a group of enzymes for which the catalytic mechanism has recently been established. These enzymes convert their substrates via the intermediate formation of a covalent enzyme-substrate complex. Interestingly, the formation of the intermediate proceeds faster by orders of magnitude than the subsequent hydrolysis, ie, the formation of the terminal product. Under normal circumstances, this does not pose a problem, since the microsomal epoxide hydrolase (mEH), the epoxide hydrolases with the best documented importance in the metabolism of carcinogens, is highly abundant in the liver, the organ with the highest capacity to metabolically generate epoxides. Computer simulation provides evidence that the high amount of mEH enzyme is favorable for the control of the steady-state level of a substrate epoxide and can keep it extremely low. However, once the mEH is titrated out under conditions of extraordinarily high epoxide concentration, the epoxide steady-state level steeply rises, leading to a sudden burst of the genotoxic effect of the noxious agent. This prediction of the computer simulation is nicely supported by experimental work. V79 Chinese hamster cells that we have genetically engineered to express human mEH at about the same level as that observed in human liver are completely protected from any measurable genotoxic effect of the model compound styrene oxide (STO) up to a dose of 100 microM in the cell culture medium (toxicokinetic threshold). In V79 cells that do not express mEH, STO leads to the formation of DNA strand breaks in a dose-dependent manner with no toxicokinetic threshold observable. Above 100 microM, the genotoxic effect of STO in the mEH-expressing cell line parallels the one in the parental cell line. Thus, the saturable protection from STO-induced strand breaks by mEH represents a typical example of a practical threshold. However, it must be pointed out that even in the presence of protective amounts of mEH, a minute but definite level of STO is present that does not contribute sufficiently to the strand break formation to overcome the background noise of the detection procedure. As pointed out above, absolute thresholds probably do not exist in chemical carcinogenesis.