Detection of extrachromosomal circular DNA sequences from tumor cells by an alkaline lysis, Alu-polymerase chain reaction technique.

Abstract

Extrachromosomal circular DNAs ranging in size from submicroscopic molecules of approximately 100 kb to cytogenetically resolvable structures of 1000+ kb called minute and double-minute chromosomes have been shown to harbor amplified genes in primary tumor cells, tumor cell lines, and drug-resistant cells grown in vitro. The presence of these molecules in transformed and malignant cells trends to reflect genetic instability and also suggests that role in tumor progression. Using a colon carcinoma cell line, we developed a technique to detect extrachromosomal circular DNA-specific sequences by Alu-polymerase chain reaction. Circular DNA was enriched by selective alkaline denaturation of genomic DNA. We have successfully performed this procedure with a minimum of 5 x 10(5) cells. The technique does not require any prior knowledge of the sequences located on the covalent circular DNA molecules for their detection. The procedure should be useful as a routine screen of primary tumor cells for the presence of extrachromosomal circular DNA and should permit the preparation of specific probes ot aid in their detailed characterizations.

• Authors: S Sen, S Rani, E J Freireich, R Hewitt, S A Stass
• Journal: Molecular carcinogenesis (Mol Carcinog) Vol. 5 Issue 2 Pg. 107-10 ( 1992) ISSN: 0899-1987 [Print] United States
• PMID: 1554408 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Biomarkers, Tumor
• DNA, Circular
• DNA, Neoplasm

Topics

• Base Sequence
• Biomarkers, Tumor
• DNA, Circular (analysis)
• DNA, Neoplasm (analysis)
• Extrachromosomal Inheritance
• Humans
• Leukemia (genetics)
• Molecular Sequence Data
• Nucleic Acid Denaturation
• Polymerase Chain Reaction (methods)