Functional characterization of the HOXB13 promoter region.
Homeobox (HOX) genes are crucial regulators of cell growth and differentiation. These genes initiate and control gene expression cascades that drive development. More recently, the absent or aberrant expression of HOX genes has been implicated in cancer development. Despite the observance of these expression changes, the regulation of the HOX genes in adult tissues and how these genes become deregulated in cancerous tissues still needs much investigation. We characterized the promoter region of the HOXB13 gene. A 3 kb region upstream of the HOXB13 gene, which included the 5'UTR, increased reporter gene expression in LNCaP cells by approximately 99 fold over the promoterless control construct. A highly conserved 179 base pair fragment containing only the 5'UTR of the HOXB13 gene constituted a minimal promoter in the LNCaP cell line. Strong promoter activity was seen in the presence or absence of testosterone, although testosterone exposure did decrease expression in LNCaP cells by 50%. In an androgen insensitive cell line Du145, no sensitivity to testosterone was detected and a consistent low basal level of expression was observed. Since HOXB13 expression is highly tissue specific, we investigated the ability of the promoter to drive expression in tissues other than prostate. We observed highest expression in LNCaP cells with low levels of expression in lung, retinoblastoma, and colon cancer cells and higher expression in MCF7 breast cancer cells.
Cross
Deanna S
DS
Marshfield Clinic Research Foundation, 1000 N. Oak Ave, Marshfield, WI 54449-5790, USA. cross.deanna@mcrf.mfldclin.edu
Burmester
J K
JK
D020121:5' Untranslated Regions; D001483:Base Sequence; D045744:Cell Line, Tumor; D017124:Conserved Sequence; D030561:Databases, Nucleic Acid; D015972:Gene Expression Regulation, Neoplastic; D005801:Genes, Homeobox; D017930:Genes, Reporter; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013739:Testosterone / Q000494:pharmacology
D020121:5' Untranslated Regions; C484414:HOXB13 protein, human; D018398:Homeodomain Proteins; D013739:Testosterone; D008156:Luciferases