Bcl-2 and bcl-xL Antisense Oligonucleotides Induce Apoptosis in Melanoma Cells of Different Clinical Stages

Recent clinical studies have shown the promise of bcl-2 antisense therapy in patients with melanoma. To further demonstrate the importance of bcl-2 and validate the related antiapoptotic protein bcl-xL as targets for antisense therapy in melanoma, their implication as survival factors in melanoma ce...

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Bibliographic Details
Published in:Journal of investigative dermatology 2002-03, Vol.118 (3), p.505-512
Main Authors: Olie, Robert A., Hafner, Christoph, Küttel, Renzo, Sigrist, Brigitte, Stahel, Rolf A., Zangemeister-Wittke, Uwe, Willers, Jörg, Dummer, Reinhard, Hall, Jonathan
Format: Article
Language:English
Subjects:
antisense
apoptosis
Apoptosis - physiology
bcl-2
bcl-X Protein
bcl-xL
Biological and medical sciences
Cell Division - physiology
Cell Survival - physiology
Dermatology
Down-Regulation - physiology
Gene Expression - drug effects
Genes, bcl-2 - genetics
Humans
Medical sciences
Melanocytes - metabolism
melanoma
Melanoma - pathology
Melanoma - physiopathology
Neoplasm Staging
Oligonucleotides, Antisense - pharmacology
Proto-Oncogene Proteins c-bcl-2 - genetics
Proto-Oncogene Proteins c-bcl-2 - metabolism
Reference Values
Tumor Cells, Cultured
Tumors of the skin and soft tissue. Premalignant lesions
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Summary:Recent clinical studies have shown the promise of bcl-2 antisense therapy in patients with melanoma. To further demonstrate the importance of bcl-2 and validate the related antiapoptotic protein bcl-xL as targets for antisense therapy in melanoma, their implication as survival factors in melanoma cells of different clinical stages as well as in normal melanocytes was investigated. Primary cell cultures derived from 17 melanomas, the cell line A375, and normal melanocytes from healthy donors were treated with antisense oligonucleotides targeting either the bcl-xL mRNA or the bcl-2 and the bcl-xL mRNAs simultaneously. Bcl-2 and bcl-xL expression in cells was analyzed by real-time polymerase chain reaction and Western blotting. Cell viability was assessed in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and apoptosis assays. Bcl-2 expression was low in melanoma cells of stages I, II, and III, hardly detectable in A375 cells, but high in normal melanocytes. Bcl-xL expression was high in all cell types tested. As shown in A375 cells and the stage III melanoma cells 0513, both the bcl-xL monospecific oligonucleotide 4259 and the bcl-2/bcl-xL bispecific oligonucleotide 4625 effectively reduced tumor cell viability by induction of apoptosis with IC50 values ranging from 200 to 350 nM. Oligonucleotide 4625 proved to be superior to 4259, as it significantly reduced the viability of cells from all melanoma stages. Both oligonucleotides reduced also the viability of normal melanocytes. Our data suggest that bcl-2 and bcl-xL are promising targets for antisense therapy of melanoma, and that the simultaneous downregulation of their expression may provide additional clinical benefit.
ISSN:0022-202X
1523-1747
DOI:10.1046/j.0022-202x.2001.01677.x