Carcinogenesis, Teratogenesis & Mutagenesis

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Site-directed analysis of key transcriptionaI regulatory regions of ITGB6 gene in oral squamous cell carcinoma cells

CHEN Xi-he1，2，DENG Xiao-ling3，*，YIN Li-qin2，FU Yu-cai2，XU Ming-yan1，*   

  1. (1. Department of Stomatology, the Second Affiliated Hospital of Shantou University Medical College, Shantou 515041; 2. Laboratory of Cellular Senescence, Shantou University Medical College, Shantou 515041; 3. Department of Cell Biology and Genetics, Shantou University Medical College, Shantou 515041, Guangdong, China)
  • Received:2012-12-20 Revised:2013-04-07 Online:2013-05-30 Published:2013-05-30
  • Contact: DENG Xiao-ling，E-mail：caseydxl@ yahoo.com；XU Ming-yan，E－mail： myxu@stu.edu.cn

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Abstract:

OBJECTIVE: To identify the key transcriptional regulatory regions in oral squamous cell carcinoma (OSCC) cell lines，and lays the groundwork for investigating the mechanism of ITGB6 transcriptional regulation in OSCC. METHODS：The recombinant pGL2 luciferase reporter constructs containing different lengths of 5' -flanking DNA fragments upstream of transcription initiation site of ITGB6 gene were constructed，and were transfected into two OSCC cell lines TCA8113 and SAS. The promoter activities were detected using dual-luciferase reporter assay system and the potential transcription factor binding sites at key transcriptional regulatory region of ITGB6 gene were predicted by bioinformatics method. RESULTS：The recombinant reporter constructs containing different lengths of 5' -flanking region of ITGB6 gene were obtained. When the length of ITGB6 5' -flanking region was reduced from -187 to -35 and -35 to +27，the transcriptional activity decreased significantly. Two potential Ets-1 binding sites and one potential IRF-4 binding site were identified in the region of -187 to +27 of ITGB6 gene. CONCLUSION：The -187/-35 and -35/+27 regions are the two key transcriptional regulatory regions of ITGB6 gene in OSCC cell lines.

Key words: oral squamous cell carcinoma, ITGB6, key regions, transcription factor, luciferase reporter assay