Regulation of cytochrome P450 gene expression in human colon and breast tumour xenografts

Abstract

It is extremely difficult to identify the factors which regulate the expression of drug-metabolising enzymes in man. To address this problem, we have developed a model involving the use of human tumours grown as xenografts in immune deficient mice. Mice bearing human colon or breast tumours as xenografts were challenged with a range of compounds, known from animal studies to be inducers of cytochrome P450s from a variety of gene families. Almost all of the compounds tested could induce human tumour P450 expression, measured either by Western blot or immunohistochemical analysis. Indeed, the levels of P450s from several distinct gene families or subfamilies including CYP2A, CYP2B, CYP2C, CYP3A and CYP4A were induced. Of particular interest was the profound induction of human P450s by 1,4 bis 2-(3,5dichloro-pyridyloxybenzene)(TCPOBOP), a compound which exhibits a marked species specificity in its ability to induce P450 expression in experimental animals. Induction of a human CYP2B protein by this compound was confirmed by Northern blot analysis and in situ hybridisation for mRNA, indicating that induction occurred at the level of transcription. These studies have a variety of implications: they provide a method for approaching the previously intractable problem of how environmental, hormonal and metabolic factors regulate human P450 genes and other genes involved in drug metabolism; they demonstrate that human tumours express P450s constitutively and that the levels of these proteins can be modulated by exogenous agents.