CCAAT/Enhancer-binding protein β regulates expression of human T1R3 taste receptor gene in the bile duct carcinoma cell line, HuCCT1

Takashi Toyono, Yuji Seta, Shinji Kataoka, Kuniaki Toyoshima
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引用次数: 18

Abstract

The T1R family (T1R1, T1R2 and T1R3 receptors) has a role in the detection of umami and sweet tastes in taste buds. Although T1R3 is also expressed in the intrahepatic bile duct, the expression patterns of T1R1 and T1R2 in this region have not been determined. In addition, the mechanisms of transcriptional regulation of the human T1R3 gene (Tas1r3) have not been elucidated. In this study, we determined the expression patterns of T1R2 and T1R3 in human liver and the function of C/EBPβ in Tas1r3 promoter activity. Immunohistochemistry showed that T1R2 and T1R3 were expressed in the intrahepatic bile duct. 5′-RACE analysis revealed that the transcriptional start sites of Tas1r3 were located 67 bp and 176 bp upstream of the ATG. Promoter analysis of Tas1r3 was performed using the luciferase reporter assay and EMSA in the Tas1r3-expressing cell line, HuCCT1. The 226-bp region upstream of the ATG had promoter activity, and C/EBPβ activated the Tas1r3 promoter activity in HuCCT1 cells. These results show that T1R2 and T1R3 receptors, in addition to their role in taste perception, may also have a role in intrahepatic cholangiocytes. C/EBPβ was identified as the transcription factor regulating Tas1r3 promoter activity in HuCCT1 cells.

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CCAAT/增强子结合蛋白β调控人T1R3味觉受体基因在胆管癌细胞系HuCCT1中的表达
T1R家族(T1R1, T1R2和T1R3受体)在味蕾中检测鲜味和甜味中起作用。虽然T1R3也在肝内胆管中表达,但T1R1和T1R2在该区域的表达模式尚未确定。此外,人类T1R3基因(Tas1r3)的转录调控机制尚未阐明。在本研究中,我们测定了T1R2和T1R3在人肝脏中的表达模式,以及C/EBPβ在Tas1r3启动子活性中的作用。免疫组化显示T1R2和T1R3在肝内胆管中表达。5 ' -RACE分析显示,Tas1r3的转录起始位点位于ATG上游67 bp和176 bp。在表达Tas1r3的细胞系HuCCT1中,使用荧光素酶报告基因法和EMSA对Tas1r3进行启动子分析。ATG上游的226 bp区域具有启动子活性,C/EBPβ激活了HuCCT1细胞中的Tas1r3启动子活性。这些结果表明,T1R2和T1R3受体除了在味觉感知中发挥作用外,还可能在肝内胆管细胞中发挥作用。C/EBPβ在HuCCT1细胞中被鉴定为调控Tas1r3启动子活性的转录因子。
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