关键词: ADAM17, 启动子, 荧光报告质粒, FoxM1

Abstract: Objective To construct a luciferase reporter plasmid of human ADAM17 gene promoter and mutate specific sites, and to identify their biological activity. Methods FoxM1 binding sites on ADAM17 promoter were analyzed by bioinformatics. The target fragment was amplified with PCR by using genomic DNA of human gastric cancer cell line MGC-803 as template, and ligated to luciferase reporter vector pGL3-basic; the three FoxM1 binding sites were mutated on the basis of recombinant plasmid; these plasmids were transferred to 293T cells and the promoter activities were determined by double luciferase reporter gene detection system. Results The sequencing results showed that the ADAM17 promoter fluorescent reporter gene and its mutants were successfully constructed. Compared with the pGL3-basic group, the luciferase activity mediated by pGL3-ADAM17(-1 365~+51) group was significantly increased (P< 0.05); Compared with the control group, the luciferase activity mediated by overexpressing FoxM1 was significantly increased (P<0.05), and the pGL3-ADAM17(-1 365~+51) group was higher than that of the three mutation groups (P<0.05) Conclusions The human ADAM17 promoter luciferase reporter gene and its mutants are successfully constructed, and the transcriptional regulation of ADAM17 by FoxM1 is preliminarily verified.

Key words: ADAM17, promoter, luciferase reporter plasmid, FoxM1