Detection and isolation of single tumor cells containing mutated DNA sequences

James F. Leary; Feng He; Lisa M. Reece

doi:10.1117/12.346730

21 April 1999 Detection and isolation of single tumor cells containing mutated DNA sequences

James F. Leary, Feng He, Lisa M. Reece

Proceedings Volume 3603, Systems and Technologies for Clinical Diagnostics and Drug Discovery II; (1999) https://doi.org/10.1117/12.346730
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

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.

Citation Download Citation

James F. Leary, Feng He, and Lisa M. Reece "Detection and isolation of single tumor cells containing mutated DNA sequences", Proc. SPIE 3603, Systems and Technologies for Clinical Diagnostics and Drug Discovery II, (21 April 1999); https://doi.org/10.1117/12.346730

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