Construction and expression of intracellular anti-ATF-1 single chain Fv fragment: a modality to inhibit melanoma tumor growth and metastasis.

Abstract

The expression of the transcriptional regulators activating transcription factor-1 (ATF-1) and cAMP-responsive element (CRE)-binding protein (CREB) is upregulated in metastatic melanoma cells. However, how overexpression of ATF-1/CREB contributes to the acquisition of the metastatic phenotype is unclear. Monoclonal antibodies against ATF-1 have previously been used to inhibit the transcriptional activity of ATF-1 and its associated proteins. Here, the effect of disrupting ATF-1 activity was investigated using intracellular expression of an inhibitory anti-ATF-1 single chain antibody fragment (ScFv). Intracellular expression of ScFv anti-ATF-1 in human melanoma cells caused significant reduction in CRE-dependent promoter activation. In addition, expression of ScFv anti-ATF-1 in melanoma cells suppressed their tumorigenicity and metastatic potential in nude mice. Furthermore, expression of ScFv anti-ATF-1 rendered the melanoma cells susceptible to thapsigargin-induced apoptosis in vitro and caused massive apoptosis in vivo in tumors transplanted subcutaneously into nude mice. These studies demonstrate the potential usage of ScFv anti-ATF-1 as an inhibitor of tumor growth and metastasis of solid tumors overexpressing ATF-1/CREB.

• Authors: Carmen Tellez, Didier Jean, Menashe Bar-Eli
• Journal: Methods (San Diego, Calif.) (Methods) Vol. 34 Issue 2 Pg. 233-9 (Oct 2004) ISSN: 1046-2023 [Print] United States
• PMID: 15312676 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S., Review)

Chemical References

• Activating Transcription Factor 1
• Antibodies, Neoplasm
• DNA-Binding Proteins
• Immunoglobulin Variable Region
• Transcription Factors

Topics

• Activating Transcription Factor 1
• Animals
• Antibodies, Neoplasm (biosynthesis, genetics, pharmacology)
• Cell Line, Tumor
• Cell Proliferation
• DNA-Binding Proteins (immunology)
• Gene Expression Regulation, Neoplastic
• Humans
• Immunoglobulin Variable Region (biosynthesis, genetics, pharmacology)
• Melanoma (immunology, pathology, prevention & control)
• Transcription Factors (immunology)