Transcription profiling of human HNF1beta mutated vs wildtype embryonic kidney cell line HEK293 to identify potential HNF1beta target genes whose activity may be deregulated in human patients

ABSTRACT: Hepatocyte nuclear factor 1beta (HNF1beta, TCF2) is a tissue-specific transcription factor whose mutation in humans leads to renal cysts, genital malformations, pancreas atrophy and maturity onset diabetes of the young (MODY5). Furthermore, HNF1beta overexpression has been observed in clear cell cancer of the ovary. To identify potential HNF1beta target genes whose activity may be deregulated in human patients we established a human embryonic kidney cell line (HEK293) expressing HNF1beta conditionally. Using Flp recombinase we introduced wildtype or mutated HNF1beta at a defined chromosomal position allowing a most reproducible induction of the HNF1beta derivatives upon tetracycline addition. By oligonucleotide microarrays we identified 25 HNF1beta regulated genes. By an identical approach we identified that the closely related transcription factor HNF1alpha (TCF1) affects only nine genes in HEK293 cells and thus is a less efficient factor in these kidney cells. The HNF1beta target genes dipeptidyl peptidase 4 (DPP4), angiotensin converting enzyme 2 (ACE2) and osteopontin (SPP1) are most likely direct target genes, as they contain functional HNF1 binding sites in their promoter region. Since nine of the potential HNF1beta target genes are deregulated in clear cell carcinoma of the ovary, we propose that HNF1beta overexpression in the ovarian cancer participates in the altered expression pattern Experiment Overall Design: Two different clones (biological replicates) were analyzed per transcription factor or mutant. By tetracyclin addition the expression of the transcription factors was induced for 24 hours prior to RNA extraction and array analysis.

ORGANISM(S): Homo sapiens

SUBMITTER: Ludger Klein-Hitpass 

PROVIDER: E-GEOD-3308 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress