Liver-specific expression of human insulin-like growth factor binding protein 1: functional role of transcription factor HNF1 in vivo

Abstract

Tissue-specific expression of insulin-like growth factor binding protein 1 (IGFBP-1) in the liver has been studied using differentiated (H4II and C2Rev7) and dedifferentiated (H5 and C2) rat hepatoma cell lines. Northern blot analysis showed that endogenous IGFBP-1 mRNA was expressed only in the differentiated cell lines. The first 341 base pairs 5' to the transcription initiation site of the human IGFBP-1 gene were inserted upstream of the chloramphenicol acetyltransferase reporter gene (pBP-1(341)). Expression of this gene from the human IGFBP-1 promoter was 10-16 times more efficient in the H4II line than in the other hepatoma cell lines and 40 and approximately 12 times more so than in rat fibroblasts (FR3T3) and a human cervical carcinoma cell line (C33), respectively. Cotransfection of pBP-1(341) and pRSV-HNF1 and/or pRSV-v-HNF1 (eukaryotic expression vectors that drive the synthesis of the liver-enriched trans-acting factor HNF1 or of v-HNF1, a related form) in C33 recipient cells yielded a 6-fold increase in IGFBP-1 promoter activity by HNF1 and a 2-fold increase by v-HNF1. These increases were dependent on the integrity of an HNF1 binding site located 58-74 nucleotides upstream of the cap site. Stimulation of promoter activity by cotransfection of both HNF1 and v-HNF1 fell between these values. Our results indicate that HNF1 is instrumental in human IGFBP-1 promoter activity in vivo and that v-HNF1 modulates this functional role.