Induction of apoptosis in human corneal and HeLa cells by mutated BIGH3.

摘要: PURPOSE: To determine the effects of overexpression mutated BIGH3 in HeLa and human corneal epithelial (HCE) cells. METHODS: Six mutations known to be responsible for autosomal dominant dystrophies linked chromosome 5 were generated a expression vector transfected HCE The secretion various BIGH3-EGFP fusion proteins measured by Western blot analysis. Apoptotic cells identified Hoechst/propidium iodide annexin V staining. Lactate dehydrogenase (LDH) activity was medium Truncated protein site-specific exact region that mediated apoptosis. RESULT: overexpressed secreted regardless its mutation status clearly observed culture medium. Overexpression induced apoptosis both cell lines through activation caspase-3. Although all disease-causing tested this experiment apoptosis, strongest effect with R124C R555W mutations. carboxyl-truncated did not induce suggesting located C-terminal domain necessary mediate death. In addition, Pro-Asp-Ile (PDI) site at 616-618 sufficient prevent induction CONCLUSIONS: induces pathway uses PDI fourth internal Fas Because DI is interaction alpha 3 beta 1 integrins, it suggests integrins play role mediating system used current study. This work key element pathophysiology BIGH3-related dystrophies.