Share Email Print
cover

Proceedings Paper

Detection and isolation of single tumor cells containing mutated DNA sequences
Author(s): James F. Leary; Feng He; Lisa M. Reece
Format Member Price Non-Member Price
PDF $14.40 $18.00

Paper Abstract

One of the problems in treating breast cancer patients is discovering the gene rearrangements that are occurring while the patient is in apparent remission. Spontaneous mutations in DNA sequences, particularly in tumor suppressor genes, can lead to the evolution of new clones of tumor cells that may be able to evade both clinical treatments and the patient's immune surveillance system. Isolation of these tumor clones is extremely difficult. Rare-event analysis and single-cell sorting techniques must be used to successfully detect and isolate these tumor clones. PCR amplification of selected gene sequences followed by TA cloning techniques can then be used to perform single-cell DNA sequencing in those gene regions. In this paper we present preliminary data showing successful detection and single-cell sorting of rare tumor clones from defined cell mixtures. Using TA cloning techniques and PCR we have been able to detect a single base-pair mutation in the PTEN tumor suppressor gene in single cells from a breast cancer cell line. Thus, while extremely difficult, it should in the future be possible to isolate tumor clones form a patient for subsequent molecular analyses of DNA mutations in critical gene regions.

Paper Details

Date Published: 21 April 1999
PDF: 9 pages
Proc. SPIE 3603, Systems and Technologies for Clinical Diagnostics and Drug Discovery II, (21 April 1999); doi: 10.1117/12.346730
Show Author Affiliations
James F. Leary, Univ. of Texas Medical Branch at Galveston (United States)
Feng He, Univ. of Texas Medical Branch at Galveston (United States)
Lisa M. Reece, Univ. of Texas Medical Branch at Galveston (United States)


Published in SPIE Proceedings Vol. 3603:
Systems and Technologies for Clinical Diagnostics and Drug Discovery II
Gerald E. Cohn; John C. Owicki, Editor(s)

© SPIE. Terms of Use
Back to Top