ausblenden:
Schlagwörter:
Antineoplastic Agents/isolation & purification/*pharmacology
Antineoplastic Combined Chemotherapy Protocols/pharmacology
*Apoptosis
Biomarkers, Tumor/genetics/*metabolism
Bystander Effect
Cell Cycle/drug effects
Cell Line, Tumor
Cell Survival/drug effects
Cycloheximide/pharmacology
Ether-A-Go-Go Potassium Channels/genetics/*metabolism
Gene Expression/drug effects
Humans
Neoplasms/drug therapy/metabolism
Receptors, TNF-Related Apoptosis-Inducing Ligand/genetics/metabolism
Recombinant Fusion Proteins/biosynthesis/isolation & purification/*pharmacology
Single-Chain Antibodies/biosynthesis/isolation & purification/*pharmacology
TNF-Related Apoptosis-Inducing Ligand/biosynthesis/isolation &
purification/*pharmacology
Zusammenfassung:
BACKGROUND: The search for strategies to target ion channels for therapeutic applications has become of increasing interest. Especially, the potassium channel K(V)10.1 (Ether-a-go-go) is attractive as target since this surface protein is virtually not detected in normal tissue outside the central nervous system, but is expressed in approximately 70% of tumors from different origins. METHODS: We designed a single-chain antibody against an extracellular region of K(V)10.1 (scFv62) and fused it to the human soluble TRAIL. The K(V)10.1-specific scFv62 antibody -TRAIL fusion protein was expressed in CHO-K1 cells, purified by chromatography and tested for biological activity. RESULTS: Prostate cancer cells, either positive or negative for K(V)10.1 were treated with the purified construct. After sensitization with cytotoxic drugs, scFv62-TRAIL induced apoptosis only in K(V)10.1-positive cancer cells, but not in non-tumor cells, nor in tumor cells lacking K(V)10.1 expression. In co-cultures with K(V)10.1-positive cancer cells the fusion protein also induced apoptosis in bystander K(V)10.1-negative cancer cells, while normal prostate epithelial cells were not affected when present as bystander. CONCLUSIONS: K(V)10.1 represents a novel therapeutic target for cancer. We could design a strategy that selectively kills tumor cells based on a K(V)10.1-specific antibody.
