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Modulation of Bcl-2 Protein Levels by an Intracellular Anti-Bcl-2 Single-Chain Antibody Increases Drug-induced Cytotoxicity in the Breast Cancer Cell Line MCF-7¹

Gene Therapy Program, University of Alabama at Birmingham, Birmingham, Alabama 35294 [A. P., J. G., C. R., J. G-N., D. T. C.], and The Burnham Institute, Cancer Research Center, La Jolla, California 92037 [J. C. R.]

Extensive experimental evidence suggests that Bcl-2 promotes cell survival by preventing the onset of apoptosis induced by a variety of stimuli. In addition, Bcl-2 expression has been correlated with resistance and poor response to chemotherapy in a number of cell types. Therefore, this protein represents a logical target for gene therapy strategies designed to achieve selective gene product ablation. In this study, we have developed an approach based upon intracellular expression of single-chain antibodies (sFvs) to achieve modulation of Bcl-2 protein levels in target cells. Using a transient expression system, we show that this intracellular anti-Bcl-2 sFv mediates specific reduction of Bcl-2 levels. This effect significantly enhances drug-mediated cytotoxicity in Bcl-2-overexpressing tumor cells, whereas transfection of the anti-Bcl-2 sFv did not affect the growth rate of the tumor cell lines. This method thus represents a novel and efficient way to selectively abrogate the activity of Bcl-2.

¹ This work was supported in part by NIH 5P30CA1314, NIAID P30A127767, NIH RO1 CA72532, NIH RO1 CA 68245, and by grants from Cure for Lymphoma Foundation, the Lymphoma Foundation of America, CaPCURE, and the University of California Breast Cancer Research Project (1RB-0093). A. P. is a research fellow of the National Cancer Institute of Canada, supported with funds provided by the Terry Fox run.

² To whom requests for reprints should be addressed, at University of Alabama at Birmingham, Gene Therapy Program, 1824 6th Avenue South, WTI 620, Birmingham, AL 35294. Phone: (205) 934-8627; Fax: (205) 975-7476; E-mail: david.curiel@ccc.uab.edu.

Received 6/ 9/97. Accepted 3/18/98.

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