Cloning and characterization of the human integrin β6 gene promoter

• Published in: PloS one 2015-03, Vol.10 (3), p.e0121439-e0121439
• Main Authors: Xu, Mingyan, Chen, Xihe, Yin, Hao, Yin, Liqin, Liu, Fan, Fu, Yucai, Yao, Jiangwu, Deng, Xiaoling
• Format: Article
• Language: English
• Subjects: Base pairs; Binding Sites; Biology; Cancer; Carcinogenesis; Carcinogens; Carcinoma, Squamous Cell - metabolism; CCAAT-Enhancer-Binding Proteins - metabolism; Cell Line, Tumor; Chromatin; Clonal deletion; Cloning; Cloning, Molecular - methods; Deoxyribonucleic acid; DNA; Gene deletion; Gene expression; Gene Expression Regulation, Neoplastic; Gene regulation; HEK293 Cells; Humans; Immunoprecipitation; Integrin beta Chains - chemistry; Integrin beta Chains - genetics; Integrin beta Chains - metabolism; Laboratories; Medical prognosis; Metastasis; Mouth Neoplasms - metabolism; Oral squamous cell carcinoma; Penicillin; Promoter Regions, Genetic; Proteins; Regulatory sequences; RNA polymerase; Senescence; Squamous cell carcinoma; Stat3 protein; STAT3 Transcription Factor - metabolism; TATA Box; Transcription factors; Transcription initiation; Transcription Initiation Site; Trends; Tumorigenesis; Wound healing
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0121439

The integrin β6 (ITGB6) gene, which encodes the limiting subunit of the integrin αvβ6 heterodimer, plays an important role in wound healing and carcinogenesis. The mechanism underlying ITGB6 regulation, including the identification of DNA elements and cognate transcription factors responsible for basic transcription of human ITGB6 gene, remains unknown. This report describes the cloning and characterization of the human ITGB6 promoter. Using 5'-RACE (rapid amplification of cDNA ends) analysis, the transcriptional initiation site was identified. Promoter deletion analysis identified and functionally validated a TATA box located in the region -24 to -18 base pairs upstream of the ITGB6 promoter. The regulatory elements for transcription of the ITGB6 gene were predominantly located -289 to -150 from the ITGB6 promoter and contained putative binding sites for transcription factors such as STAT3 and C/EBPα. Using chromatin immunoprecipitation assays, this study has demonstrated, for the first time, that transcription factors STAT3 and C/EBPα are involved in the positive regulation of ITGB6 transcription in oral squamous cell carcinoma cells. These findings have important implications for unraveling the mechanism of abnormal ITGB6 activation in tissue remodeling and tumorigenesis.