Identification of Sp1-elements in the promoter region of human homeobox gene NKX3.1

• Published in: Molecular biology reports Vol. 36; no. 8; pp. 2353 - 2360
• Main Authors: Yu, Chun-xiao, Jin, Tong, Chen, Wei-wen, Zhang, Peng-ju, Liu, Wen-wen, Guan, Heng-yun, Zhang, Ju, Liu, Qing-wei, Jiang, An-li
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.11.2009; Springer Nature B.V
• Subjects: Animal Anatomy; Animal Biochemistry; Base Sequence; Biomedical and Life Sciences; Cell Line, Tumor; Cellular biology; Gene deletion; Gene expression; Gene Expression Regulation, Neoplastic; gene overexpression; genetics; Histology; Homeobox; Homeodomain Proteins; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; homeotic genes; Humans; Life Sciences; Male; messenger RNA; metabolism; Molecular biology; Molecular Sequence Data; Morphology; Mutagenesis; Mutation; Nkx3.1 protein; Oligonucleotides; Overexpression; Plasmids; Polymerase chain reaction; promoter regions; Promoter Regions, Genetic; Promoters; Prostate cancer; prostatic neoplasms; Prostatic Neoplasms - genetics; Prostatic Neoplasms - metabolism; Protein Binding; Reporter gene; reporter genes; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sp1 protein; Sp1 Transcription Factor; Sp1 Transcription Factor - genetics; Sp1 Transcription Factor - metabolism; Transcription Factors; Transcription Factors - genetics; Transcription Factors - metabolism; Western blotting; Promoter; Homeobox gene; Sp1
• ISSN: 0301-4851; 1573-4978; 1573-4978
• DOI: 10.1007/s11033-009-9457-y

NKX3.1 is a prostate-specific homeobox gene related strongly to prostate development and prostate cancer. However, little is known about the mechanism for regulation of NKX3.1 in prostate cancer. With RT-PCR and western blot, we found that NKX3.1 expression was enhanced by over-expression of Sp1 at both the mRNA and protein levels in prostate cancer LNCaP cells. To identify the Sp1-elements in the promoter region of NKX3.1 , a 521 bp-promoter of human NKX3.1 gene containing three possible Sp1-elements was cloned into the upstream of the luciferase reporter gene in pGL 3 -basic plasmid. With deletion mutation analysis, plasmid construction, EMSA and oligonucleotide decoy technique, two Sp1-elements which located between +29 to +43 and −60 to −46 of NKX3.1 gene were identified and proven to be functional elements. It will be important to further study on the functions and the regulatory mechanisms of Sp1 element in NKX3.1 gene expression.