Analysis of clonality by amplification of short tandem repeats. Carcinomas of the female reproductive tract.
The clonal composition of cancers of the female reproductive tract was evaluated by analysis of patterns of X-chromosome inactivation. Using DNA extracted from frozen tissues or paraffin-embedded archival specimens as template, polymerase chain reaction (PCR) was performed to generate amplified DNA fragments of exon 1 of the X-linked androgen receptor gene, which contains a highly polymorphic trinucleotide repeat. Predigestion of tumor DNA with methylation-sensitive restriction endonuclease Hha I or Hpa II permitted selective PCR amplification from the methylated (uncleaved) allele. Of a total of 54 tumors analyzed, 50 cases showed heterozygosity (93%) and were therefore informative for clonal analysis. Monoclonal composition of the tumors was suggested in a total of 49 of 50 cases, including 12 adenocarcinomas of the uterine endometrium, 13 squamous cell carcinomas of the uterine cervix, 6 adenocarcinomas of the uterine endocervix, and 18 epithelial tumors of the ovary. However, polyclonal composition was observed in one mucinous carcinoma of the ovary, in which we previously showed that both GGT-->GAT and GGT-->GTT mutations are present in > 20% of total K-ras copies in the tissue. Our studies demonstrate the utility of PCR amplification of highly polymorphic repetitive sequences for analysis of patterns of X-chromosome inactivation. This approach is practical for the analysis of clonal cell composition in a high proportion of both formalin-fixed and frozen archival tissues.
Enomoto
T
T
Department of Radiation Biology, Osaka University Faculty of Medicine, Japan.
Fujita
M
M
Inoue
M
M
Tanizawa
O
O
Nomura
T
T
Shroyer
K R
KR
D002277:Carcinoma / Q000235:genetics / Q000473:pathology; D004303:Dosage Compensation, Genetic; D016889:Endometrial Neoplasms / Q000235:genetics / Q000473:pathology; D005091:Exons; D005260:Female; D005833:Genital Neoplasms, Female / Q000235:genetics / Q000473:pathology*; D006801:Humans; D010051:Ovarian Neoplasms / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011944:Receptors, Androgen / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000473:pathology
D011944:Receptors, Androgen