Cloning and characterization of the ADAMTS14 promotor

ADAMTS genes have been shown to play an important role in the emergence, development and metastasis of tumors. ADAMTS14, one of the members of this gene family, forms collagen ADAMTS and is involved in collagen processing. ADAMTS14 was discovered in 2002 as a gene encoding a proteinase located on human chromosome 10q22. There are studies that aberrant expression of ADAMTS14 affects tumor cell behaviors, including cell proliferation and metastasis. With in the scope of this master thesis, the expression of ADAMTS14 in different cell lines was investigated. In addition, cloning and validation of 5' deletion mutants created from the promoter region of the ADAMTS14 gene were performed. For confirmation, transient transfection with calcium phosphate precipitation was performed in the SW480 colon cancer cell line. In order to examine the relationship between ADAMTS14 gene expression and IL-6, IL-6 cytokine was applied to the cells in different amounts (20 and 40ng), and its effects were determined at the mRNA level by Real-Time PCR method.