The Structural And Functional Studies Of Sipa1 Gene Promoter Sequence And It's SNPs

SIPA1 protein, signaling-induced proliferating-associating protein 1, is a Rap1 GTPase activating protein(Rap1GAP). It can catalyze Rap1 protein from active Rap1 GTP to inactive Rap1 GDP and then regulate Rap1 protein relative function for cell adhesion, cellular kinase activity and cell nuclear gene regulation. The gene of SIPA1 is localized in chromosome 11 and it consist of 16 exons. The SIPA1 protein itself contains 1042 amino acids.Invasive breast cancer is the most diagnosed cancers for women. It is crucial for patients survival to make early diagnosis and precise treatment. It has been reported that SIPA1 protein was closely related to the breast cancer pathogenesis, proliferation, adhesion, invasion, metastasis. Simultaneously, some researchers found out that SIPA1 protein was associated with metastasis and invasion of colorectal cancer, breast cancer, melanoma, prostate cancer and cervical cancer. While the relationship between SIPA1 protein and many cancers is obvious, some research on clinic found that the SNPs, rs931127 and rs3741378, on Sipa1 gene was tightly associated with cervical metastasis, the survival and development of non-small cell lung cancer and breast cancer.The function of SIPA1 protein is so powerful in cancers that it is necessary to know how it is regulated. Little is known about these respects research. So my research on master stage is to explore the structure and function of Sipa1 gene. The following is my research conclusion and results.First, found out firstly the location of Sipa1 gene promoter and it's promoter core region by predicting and constructing the different region of Sipa1 gene promoter and Dual-Luciferase Reporter Assay System.Second, confirmed that the SNP, rs931127, on Sipa1 gene promoter don't impact the activity of Sipa1 gene promoter by specific-mutation and Dual-Luciferase Reporter Assay System.Third, confirmed that the SNP, rs3741378, on Sipa1 gene exon 3 can significantly change the expression of SIPA1 protein by specific-mutation( pcDNA3-flag-SIPA1 C ?pcDNA3-flag-SIPA1 T)and Western blotting.Fourth, by collecting different cancer cell lines genome and sequencing the SNPs including rs931127 and rs3741378 genotype and by cell nuclear and cytoplasmic assay, then found that the SIPA1 protein cell nuclear sublocalization is not significantly associated with the SNP genotype of Sipa1 gene.In conclusion, this research successively found the promoter of Sipa1 gene and it's promoter core region. It revealed the structure of Sipa1 gene and provided the foundation for seeking for the mechanism that SIPA1 how to be regulated and the relative transcription factors. Moreover, identify the SNPs of Sipa1 gene function influence for the SIPA1 expression.