Transcriptional activation of the porcine P450 11A insulin-like growth factor response element in MCF-7 breast cancer cells

Abstract

Insulin-like growth factor-I (IGF-I) stimulates the growth of MCF-7 breast cancer cells. This study determined the transcriptional activity of an IGF-I-responsive region (IGFRE) of porcine P450 11A (P450scc) after transfection into MCF-7 cells. IGF-I induced transcriptional activity of a porcine P450scc core promoter luciferase construct containing the IGFRE transfected in MCF-7 cells. Electrophoretic mobility shift assay with nuclear protein extract from MCF-7 cells showed two transcription factors binding to the IGFRE. Supershift assay determined that one transcription factor was Sp1. Electrophoretic mobility shift assay and transfection experiments with selected mutations to the IGFRE found that binding of both transcription factors was necessary to confer an IGF-I response. The binding activity of both transcription factors was increased with IGF-I treatment. In conclusion, MCF-7 cells contain Sp1 and another unknown transcription factor, P2, that bind to a known IGFRE (porcine P450scc) and induce reporter gene transcriptional activity with IGF-I treatment. Because Sp1 is a ubiquitous transcription factor, determining the identity of P2 may lead to cell- specific methods to impair breast cancer cell growth as mediated by IGF-I.