Transcription factor GRHL3 regulates the expression of GPR108 gene and its mechanism

doi:10.3969/j.issn.1006-7795.2022.03.019

Abstract

Abstract: Objective To study the molecular mechanism of transcription factor grainyhead-like (GRHL3) involved in the regulation of GPR108 expression. Methods Bioinformatics was used to analyze the expression level and relationship of GRHL3 and G protein-coupled receptor 108(GPR108) in various cancer cells. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of GRHL3 and GPR108 genes in human epidermoid carcinoma cell line (A-431), human hepatocellular liver carcinoma cell line (Hep G2) and human neuroblastoma cell line (SH-SY5Y). The luciferase reporter gene expression vector containing GPR108 promoter and mutant GPR108 promoter was constructed by molecular cloning technology. The regulatory effect of GRHL3 on GPR108 promoter was observed by double luciferase reporter gene detection system; The relationship between GRHL3 and GPR108 promoter binding sites was analyzed by chromatin immunoprecipitation assay (ChIP). Results Bioinformatics analysis showed that the expression of GRHL3 was lower and GPR108 was higher in various cancer cells. The luciferase reporter gene expression vectors of GPR108 promoter and mutant GPR108 promoter were successfully constructed by molecular cloning technology. Luciferase results showed that GRHL3 significantly inhibited the promoter of GPR108. After deleting the binding site on the GPR108 promoter, the negative regulatory effect of GRHL3 on the GPR108 promoter was weakened; ChIP assay showed that GRHL3 could interact with GPR108 promoter binding site. Conclusion The transcription factor GRHL3 can be negatively regulated by binding to the conserved sequence 5′-AACCGGTG-3′ on the GPR108 promoter.

Key words: grainyhead-like (GRHL3), G protein-coupled receptor 108 (GPR108), promoter

CLC Number:

Ni Mingyue, Liu Yuanli, Tu Zhenzhen, Guo Liyu, Zang Dandan, Zhou Haisheng. Transcription factor GRHL3 regulates the expression of GPR108 gene and its mechanism[J]. Journal of Capital Medical University, 2022, 43(3): 446-455.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2022.03.019

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2022/V43/I3/446

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