Analysis of the aromatase cytochrome P450 gene in human breast cancers.
Aromatase activity may be detected in most, but not all, breast cancers, and in certain tumours there appears to be decreased sensitivity to the aromatase inhibitor 4-hydroxyandrostenedione (4-OHA). The aims of the present study were to measure aromatase activity, and its sensitivity to 4-OHA, in breast tumours, and to examine the CYP19 gene encoding the aromatase cytochrome P450 (P450arom) for the presence of mutations. In vitro aromatase activity and sensitivity to 4-OHA were measured by determining the conversion of tritiated testosterone to tritiated oestradiol in breast tumour tissue in the absence and presence of 4-OHA (10 nM). Genomic DNA was extracted from five tumours: one showing no detectable aromatase activity and four displaying evidence of aromatase activity (two sensitive and two insensitive to 4-OHA). Subsequent PCR-single-strand conformation polymorphism analysis revealed a variation in the mobility of single-stranded DNA for exons III, VII and X, corresponding, as shown by direct sequencing of PCR products, to common polymorphism of the aromatase gene. This study does not provide evidence for mutation in the coding exons of the P450arom gene which would account for either the absence of aromatase activity or its changed sensitivity to 4-OHA in breast cancers.
Sourdaine
P
P
ICRF Medical Oncology Unit, Western General Hospital, Edinburgh, UK.
Parker
M G
MG
Telford
J
J
Miller
W R
WR
D000735:Androstenedione / Q000031:analogs & derivatives / Q000494:pharmacology; D001141:Aromatase / Q000235:genetics* / Q000378:metabolism; D047072:Aromatase Inhibitors; D001483:Base Sequence; D001943:Breast Neoplasms / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D015342:DNA Probes; D004958:Estradiol / Q000096:biosynthesis; D005091:Exons / Q000235:genetics; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010920:Placenta / Q000201:enzymology; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational
D047072:Aromatase Inhibitors; D015342:DNA Probes; D000735:Androstenedione; D004958:Estradiol; D001141:Aromatase; C014594:formestane