Use of Transcriptional-Enhancer Elements Responsive to Ras, Protein Kinase A and Protein Kinase C to Evaluate Inhibitors of Specific Signal Transduction Pathways

To evaluate ras-mediated signal transduction, we constructed a transient transfection assay system that measures chloramphenicol acetyl transferase activity expressed under the transcriptional-enhancer elements responsive to ras, protein kinase C, and protein kinase A in NIH3T3 cells. Characterizati...

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Bibliographic Details
Published in:Biological & pharmaceutical bulletin 1994/06/15, Vol.17(6), pp.779-783
Main Authors: KANBE, Toshie, TSUCHIYA, KayokoS, HORI, Makoto, YAGINUMA, Satoshi, MURAKAMI, Yuko, MIZUNO, Satoshi, UEHARA, Yoshimasa
Format: Article
Language:English
Subjects:
3T3 Cells
Animals
anthracycline
Antibiotics, Antineoplastic - pharmacology
Antineoplastic agents
Base Sequence
Biological and medical sciences
Cell Line, Transformed
Chloramphenicol O-Acetyltransferase - metabolism
Cyclic AMP-Dependent Protein Kinases - metabolism
Doxorubicin - analogs & derivatives
Doxorubicin - pharmacology
Enhancer Elements, Genetic - drug effects
Gene Expression - drug effects
General aspects
Medical sciences
Mice
Molecular Sequence Data
Pharmacology. Drug treatments
Plasmids
Promoter Regions, Genetic
Protein Kinase C - metabolism
ras
ras Proteins - drug effects
ras Proteins - genetics
ras Proteins - metabolism
Signal Transduction - drug effects
Signal Transduction - genetics
signal transduction pathway
Transcription Factors - drug effects
Transcription Factors - genetics
transcriptional enhancer element
Transfection
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Summary:To evaluate ras-mediated signal transduction, we constructed a transient transfection assay system that measures chloramphenicol acetyl transferase activity expressed under the transcriptional-enhancer elements responsive to ras, protein kinase C, and protein kinase A in NIH3T3 cells. Characterization of the assay system with several known activators and inhibitors of signal transduction pathways proved that our system could reliably evaluate agents that affect individual pathways. Pirarubicin ((2"R)-4'-tetrahydropyranyladriamycin, THP), which has recently been found able to reverse ras-transformed cells, appeared to selectively inhibit the ras signal transduction pathway.
ISSN:0918-6158
1347-5215
DOI:10.1248/bpb.17.779