Bioinformatics analysis of Wnt/β-catenin pathway related regulatory elements in human mesothelin gene and identification of promoter in ovarian cancer cells / 临床与实验病理学杂志

ABSTRACT

Purpose To analyze the Wnt/β-catenin signaling pathway related TCF/LEF binding sites in human mesothelin gene and to identify the core promoter region of the gene in the ovarian cancer cells. Methods The possible TCF/LEF transcription factor binding sites in the promoter region of human meosothelin gene were analyzed by bioinformatics method. The 1764 bp promoter sequence near the 5'end of the human mesothelin gene were cloned. The fragments was truncated differently at the 5' end and cloned into p GL3-basic report gene vector and transfected into human ovarian cancer cell lines SKOV-3 and 3-AO. The activity of diffetent promoter fragmentis was detected by double luciferase reporter gene system. Results There were multiple potential TCF/LEF binding sites in the promoter region of the human mesothelin gene. Three fragments of mesothelin gene promoter region-1456-+ 308、-164-+ 308、+ 47-+ 308 were cloned and amplified successfully, the p GL3 vector was constructed by sequencing. After transfection of SKOV-3 and 3-AO cells, the double luciferase reporter gene system showed that the-1456-+ 308 fragments and-164-+ 308 framents had high promoting activity in both cell lines, and the activity of + 47-+308 fragments was significantly lower than that of the former two cell lines (P<0.01). Conclusion The-164-+ 47 sequence containing TCF/LEF transcription factor binding site is the core promoter region of mesothelin gene in ovarian cancer, which lays a foundation for further study on the regulation mechanism of mesothelin gene expression in ovarian cancer.