Calcium carbonate nanoparticle delivering vascular endothelial growth factor-C siRNA effectively inhibits lymphangiogenesis and growth of gastric cancer in vivo

A nonviral gene carrier, calcium carbonate (CaCO3) nanoparticle, was evaluated for efficient in vitro and in vivo delivery of small interfering RNA (siRNA) targeting vascular endothelial growth factor-C (VEGF-C). The chemically synthesized CaCO3 nanoparticle has a 58 nm diameter and +28.6 mV positive surface charge. It is capable of forming a CaCO3 nanoparticle–DNA complex and transferring DNA into targeted cells with high transfection efficiency while effectively protecting the encapsulated DNA from degradation. Furthermore, the CaCO3 nanoparticle–DNA complex has no obvious cytotoxicity for SGC-7901 cells, while a liposome–DNA complex exhibited measurable cytotoxicity. SGC-7901 cells transfected with a VEGF-C-targeted siRNA via CaCO3 nanoparticle exhibit significantly reduced VEGF-C expression as measured by real-time PCR and enzyme-linked immunosorbent assay; whereas no decrease in VEGF-C expression is observed in cells treated by control transfection. Transfection of SGC-7901 cells with VEGF-C siRNA via CaCO3 nanoparticle also dramatically suppresses tumor lymphangiogenesis, tumor growth and regional lymph-node metastasis in subcutaneous xenografts. Significant downregulation of VEGF-C messenger RNA expression in a subcutaneous xenograft derived from VEGF-C siRNA-treated SGC-7901 cells was confirmed by real-time PCR as compared to controls. We conclude that CaCO3 nanoparticle is a novel and nonviral system for effective delivery of siRNA for cancer gene therapy.