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. 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.
Showa College of Pharmaceutical Sciences, Tokyo, Japan.
T
T
Kanbe
K S
KS
Tsuchiya
M
M
Hori
S
S
Yaginuma
Y
Y
Murakami
S
S
Mizuno
Y
Y
Uehara
D016475:3T3 Cells; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D001483:Base Sequence; D002461:Cell Line, Transformed; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004317:Doxorubicin / Q000031:analogs & derivatives / Q000494:pharmacology; D004742:Enhancer Elements, Genetic / Q000187:drug effects*; D015870:Gene Expression / Q000187:drug effects; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011493:Protein Kinase C / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects* / Q000235:genetics; D014157:Transcription Factors / Q000187:drug effects / Q000235:genetics*; D014162:Transfection; D018631:ras Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism
D000903:Antibiotics, Antineoplastic; D014157:Transcription Factors; D004317:Doxorubicin; C027260:pirarubicin; D015500:Chloramphenicol O-Acetyltransferase; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D018631:ras Proteins