Big-h3내 RGD-motif의 유전자 변이 유도에 의한 유방암 세포의 성장 및 이동성에 미치는 영향

Site-Directed Mutagenesis of RGD-Motif in Big-h3 and its Effect on the Growth and Invasion of Breast Cancer Cells

Extracellular matrix(ECM) plays an important role in cell migration in pathological conditions such as inflammation and cancer. Big-h3 (also named transforming growth factor β induced protein, betaig-h3, Bigh3, TGF-b-induced protein h3, keratoepithelin, and RGD-CAP) is an secretory extracellular protein and includes YH and RGD motif, which interact with extracellular matrix. In this study, big-h3 and big-h3 fragment (big-h3-f, 182 amino acid.) containing RGD-motif were cloned. To determine whether RGD-motif would affect cell growth, RGD-motif with aspatic acid to RGE-motif with glutamic acid of wild-type big-h3 or big-h3-f was mutated by site-directed mutagenesis. Big-h3, big-h3-f, mutant big-h3, or mutant big-h3-f was transfected into human breast cancer cell line MCF-7 and their expression were confirmed by immunoblotting. Wild type big-h3 and big-h3-f containing RGD-motif led to increased cell growth and invasion, compared with mutant big-h3 and mutant big-h3-f containing RGE-motif. Thus, RGD-motif of big-h3 may play a major role in cell growth and invasion regulation in regard to cell-extracellular matrix interaction.