Immunohistological examination of the inter- and intracellular distribution of O6-alkylguanine DNA-alkyltransferase in human liver and melanoma.

2.50
Hdl Handle:
http://hdl.handle.net/10541/108143
Title:
Immunohistological examination of the inter- and intracellular distribution of O6-alkylguanine DNA-alkyltransferase in human liver and melanoma.
Authors:
Lee, Siow Ming; Rafferty, Joseph A; Elder, Rhoderick H; Fan, Chun-Yang; Bromley, Michael; Harris, Martin; Thatcher, Nick; Potter, P M; Altermatt, H J; Perinat-Frey, T; Cerny, T; O'Connor, Peter J; Margison, Geoffrey P
Abstract:
The tissue and cellular distribution of the DNA repair protein O6-alkylguanine-DNA-alkyltransferase (ATase) is an important question in relation to the response of tumour and normal tissues to chemotherapeutic regimes employing alkylating agents such as methyltriazenes and nitrosoureas. In order to examine this issue by immunostaining, we have raised a rabbit antiserum to apparently pure recombinant human enzyme. The antiserum is highly specific and sensitive, detecting a band at 24 kDa on western blots of crude extracts of ATase-expressing human lymphoblastoid cells, liver and melanoma. Adjacent sections of acetone or formalin fixed normal human liver and subcutaneous malignant melanoma were reacted with preimmune serum or antiserum and an immunoperoxidase detection system with silver enhancement was used to locate binding of the primary antibody to the antigen. In sections reacted with preimmune serum or with antigen-preadsorbed antiserum, only faint cytoplasmic and little or no nuclear staining was seen. In contrast, using antiserum, the reaction in positively staining cells was very intense and predominantly nuclear. In the liver, there was interindividual variation in the cellular distribution of reaction with staining present in all discernable cell types in most samples but confined to the hepatocytes and bile duct epithelial cells in others. In the melanoma sections, all discernable cell types showed mainly nuclear staining: the intensity of staining varied between tissue samples and there was evidence of a range of intermediate staining intensities with some melanoma cells showing no detectable reaction.
Affiliation:
CRC Department of Carcinogenesis, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, UK.
Citation:
Immunohistological examination of the inter- and intracellular distribution of O6-alkylguanine DNA-alkyltransferase in human liver and melanoma. 1992, 66 (2):355-60 Br J Cancer
Journal:
British Journal of Cancer
Issue Date:
Aug-1992
URI:
http://hdl.handle.net/10541/108143
PubMed ID:
1503911
Type:
Article
Language:
en
ISSN:
0007-0920
Appears in Collections:
All Christie Publications ; All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorLee, Siow Mingen
dc.contributor.authorRafferty, Joseph Aen
dc.contributor.authorElder, Rhoderick Hen
dc.contributor.authorFan, Chun-Yangen
dc.contributor.authorBromley, Michaelen
dc.contributor.authorHarris, Martinen
dc.contributor.authorThatcher, Nicken
dc.contributor.authorPotter, P Men
dc.contributor.authorAltermatt, H Jen
dc.contributor.authorPerinat-Frey, Ten
dc.contributor.authorCerny, Ten
dc.contributor.authorO'Connor, Peter Jen
dc.contributor.authorMargison, Geoffrey Pen
dc.date.accessioned2010-07-22T09:10:33Z-
dc.date.available2010-07-22T09:10:33Z-
dc.date.issued1992-08-
dc.identifier.citationImmunohistological examination of the inter- and intracellular distribution of O6-alkylguanine DNA-alkyltransferase in human liver and melanoma. 1992, 66 (2):355-60 Br J Canceren
dc.identifier.issn0007-0920-
dc.identifier.pmid1503911-
dc.identifier.urihttp://hdl.handle.net/10541/108143-
dc.description.abstractThe tissue and cellular distribution of the DNA repair protein O6-alkylguanine-DNA-alkyltransferase (ATase) is an important question in relation to the response of tumour and normal tissues to chemotherapeutic regimes employing alkylating agents such as methyltriazenes and nitrosoureas. In order to examine this issue by immunostaining, we have raised a rabbit antiserum to apparently pure recombinant human enzyme. The antiserum is highly specific and sensitive, detecting a band at 24 kDa on western blots of crude extracts of ATase-expressing human lymphoblastoid cells, liver and melanoma. Adjacent sections of acetone or formalin fixed normal human liver and subcutaneous malignant melanoma were reacted with preimmune serum or antiserum and an immunoperoxidase detection system with silver enhancement was used to locate binding of the primary antibody to the antigen. In sections reacted with preimmune serum or with antigen-preadsorbed antiserum, only faint cytoplasmic and little or no nuclear staining was seen. In contrast, using antiserum, the reaction in positively staining cells was very intense and predominantly nuclear. In the liver, there was interindividual variation in the cellular distribution of reaction with staining present in all discernable cell types in most samples but confined to the hepatocytes and bile duct epithelial cells in others. In the melanoma sections, all discernable cell types showed mainly nuclear staining: the intensity of staining varied between tissue samples and there was evidence of a range of intermediate staining intensities with some melanoma cells showing no detectable reaction.en
dc.language.isoenen
dc.subject.meshBiopsy-
dc.subject.meshBlotting, Western-
dc.subject.meshCell Line-
dc.subject.meshHumans-
dc.subject.meshImmune Sera-
dc.subject.meshImmunohistochemistry-
dc.subject.meshLiver-
dc.subject.meshMelanoma-
dc.subject.meshMethyltransferases-
dc.subject.meshO(6)-Methylguanine-DNA Methyltransferase-
dc.titleImmunohistological examination of the inter- and intracellular distribution of O6-alkylguanine DNA-alkyltransferase in human liver and melanoma.en
dc.typeArticleen
dc.contributor.departmentCRC Department of Carcinogenesis, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, UK.en
dc.identifier.journalBritish Journal of Canceren
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