title: Heterogeneity of O6-alkylguanine DNA-alkyltransferase expression in human breast tumours.
abstract: An important determinant of cellular resistance to chemotherapeutic O6-alkylating agents, which comprise methylating and chloroethylating agents, is the ability of cells to repair alkylation damage at the O6-position of guanine in DNA. This is achieved by a specific DNA repair enzyme O6-alkylguanine DNA-alkyltransferase. In this study O6-alkylguanine DNA-alkyltransferase expression was measured in human breast tumours using both biochemical and immunohistochemical techniques. O6-alkylguanine DNA-alkyltransferase activity was then compared with known clinical prognostic indices to assess the potential role of O6-alkylguanine DNA-alkyltransferase in predicting the behaviour of this common malignancy. The application of both biochemical and immunohistochemical techniques was feasible and practical. Most breast tumours expressed high levels of O6-alkylguanine DNA-alkyltransferase. Immunohistochemical analysis showed marked variation in expression not only between individuals but also within individual tumours, and in the same patient, between metastases and between primary tumour and metastatic site. O6-alkylguanine DNA-alkyltransferase activity in tissue extracts significantly correlated not only with immunohistochemical staining intensity determined by subjective quantitation, but also with measures of protein levels using a computerised image analysis system including mean grey (P<0.001), percentage of cells positive for O6-alkylguanine DNA-alkyltransferase (P<0.001), and integrated optical density (P<0.001). O6-alkylguanine DNA-alkyltransferase expression did not correlate with any of the established clinical prognostic indicators for current treatment regimens. However, immunohistochemical offers a rapid and convenient method for assessing potential utility of O6-alkylating agents or O6-alkylguanine DNA-alkyltransferase inactivating agents in future studies of breast cancer treatment.
authors.affiliation: Cancer Research UK Department of Medical Oncology, Christie Hospital, Wilmslow Road, Manchester M20 4BX, UK.
authors.forename: M J
authors.initials: MJ
authors.lastname: Clemons
authors.forename: M C
authors.initials: MC
authors.lastname: Bibby
authors.forename: H
authors.initials: H
authors.lastname: El Teraifi
authors.forename: G
authors.initials: G
authors.lastname: Forster
authors.forename: J
authors.initials: J
authors.lastname: Kelly
authors.forename: S
authors.initials: S
authors.lastname: Banerjee
authors.forename: B
authors.initials: B
authors.lastname: Cadman
authors.forename: W D J
authors.initials: WD
authors.lastname: Ryder
authors.forename: A
authors.initials: A
authors.lastname: Howell
authors.forename: G P
authors.initials: GP
authors.lastname: Margison
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D001943:Breast Neoplasms / Q000201:enzymology / Q000235:genetics* / Q000473:pathology / Q000601:surgery; D005260:Female; D006801:Humans; D008207:Lymphatic Metastasis; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000235:genetics / Q000378:metabolism*
chemical_list: D019853:O(6)-Methylguanine-DNA Methyltransferase; D000251:Adenosine Triphosphatases