Clonal X-inactivation analysis of human tumours using the human androgen receptor gene (HUMARA) polymorphism: a non-radioactive and semiquantitative strategy applicable to fresh and archival tissue.

Details

Serval ID
serval:BIB_03EEE7E1F022
Type
Article: article from journal or magazin.
Collection
Publications
Title
Clonal X-inactivation analysis of human tumours using the human androgen receptor gene (HUMARA) polymorphism: a non-radioactive and semiquantitative strategy applicable to fresh and archival tissue.
Journal
Molecular and cellular probes
Author(s)
Kopp P., Jaggi R., Tobler A., Borisch B., Oestreicher M., Sabacan L., Jameson J.L., Fey M.F.
ISSN
0890-8508 (Print)
ISSN-L
0890-8508
Publication state
Published
Issued date
06/1997
Peer-reviewed
Oui
Volume
11
Number
3
Pages
217-228
Language
english
Notes
Publication types: Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
Assessment of clonality of cellular proliferations is important in experimental and clinical cancer research. X-chromosome inactivation studies are widely used to assess clonality, but most assays require relatively large amounts of high molecular weight DNA. Two PCR-based strategies, the phosphoglycerate kinase (PGK) and the human androgen receptor (HUMARA) clonality assays allow studies of small tissue samples. The HUMARA assay was adapted to non-radioactive analysis taking advantage of an automated sequencer providing high resolution of alleles and immediate quantitation. This assay was validated by comparison with X-inactivation patterns obtained by Southern analysis with the probes M27 beta and PGK. Fifteen gastrointestinal carcinomas, 25 benign goiter nodules and normal peripheral leukocytes of 27 individuals (12 who were under 15 years and 15 over 80 years) were analysed. Furthermore, DNA extracted from formalin-fixed paraffin-embedded tissue (FPT) was analysed with the two PCR-based methods and compared with X-inactivation patterns determined by Southern analysis of high molecular weight (HMW) DNA. This modified HUMARA assay is reliable in most patients; as with other clonality assays, constitutive skewing in normal tissue precludes clonal analysis in some individuals. Extremely skewed X-inactivation patterns were found in normal peripheral leukocytes of 7 out of 15 old females (over 80 years) and in 1 of 12 of the young females tested (under 15 years). Comparison of results obtained with HMW and FPT DNA yielded consistent results for the HUMARA assay whereas the PGK PCR assay was much less reliable. The HUMARA assay thus permits studies of selected areas of tissue sections without significant stromal components, allowing correlation of histological and genotype findings in fresh and archival specimens.
Keywords
Adolescent, Aged, Aged, 80 and over, Blotting, Southern, Child, Child, Preschool, DNA Primers, DNA Probes, DNA, Neoplasm/analysis, Dosage Compensation, Genetic, Female, Gastrointestinal Neoplasms/genetics, Gastrointestinal Neoplasms/pathology, Humans, Phosphoglycerate Kinase/genetics, Polymerase Chain Reaction, Polymorphism, Genetic, Receptors, Androgen/genetics
Pubmed
Web of science
Create date
30/12/2020 15:39
Last modification date
31/12/2020 6:26
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