Electronic Thesis and Dissertation Repository

Degree

Master of Science

Program

Physiology

Supervisor

Dr. Rennian Wang

Abstract

Interactions between Stem Cell Factor (SCF) and c-Kit are critical in maintaining β-cell survival and function. VEGF-A, essential in facilitating islet vasculature formation, is secreted by β-cells. This work investigates the mechanisms by which c-Kit regulates VEGF-A production in β-cells, using a rat insulinoma (INS-1) cell line as an in vitro model. Cells were treated with recombinant hSCF or cultured with c-Kit (r) siRNA for 72 h. VEGF-A accumulation in the culture medium was measured by ELISA, while qRT-PCR, western blot, and immunofluorescence analyses were also performed. SCF augmented VEGF-A production in a time-dependent (p < 0.05 vs. control at 6 h and 24 h), and dose-dependent manner (p < 0.01 vs. control at 30-100 ng/mL). SCF stimulation increased phosphorylation of proteins along the PI3K/Akt/mTOR pathway (p < 0.05 vs. control groups), and decreased phosphorylation following siRNA treatment. Our results indicate that c-Kit exerts a regulatory effect on VEGF-A production in β-cells.


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