The Application Of ShRNA Targeting VEGF And VEGFR2 To Anticancer Research

Angiogenesis is an essential requirement for the development, progression and metastasis of malignant tumors. Clinical symptoms are only emerged after angiogenesis in many cancer patients. So how to therapeutically inhibit such angiogenesis might be exploited as a novel strategy of cancer treatment. One of the most important angiogenic factor is vascular endothelial growth factor (VEGF), which binds with high-affinity to tyrosine kinase receptors. VEGF plays a key role in tumor cell activation (autocrine), in addition to paracrine activations whereby it regulates endothelial cell functions and subsequent neovascular development. RNAi technology is currently being evaluated not only as an extremely powerful instrument for functional genomic analyses, but also as a potentially useful method to develop highly specific gene-silencing therapeutics. In our study, we designed the specific interference sequences targeting VEGF and VEGFR2, and also investigated inhibition effect of expression of VEGF or VEGFR2 in human fibrosarcoma and prostate cancer models.1. Construction and anticancer efficacy of stable shRNA expression vector targeting VEGF in human fibrosarcoma HT1080 cells.Non-viral shRNA expression vector has highly safety, but the transfection efficiency is poor. High dosage of plasmid DNA can induce interferon response after transfection into cells. Therefore, we designed a shRNA expression vector pCD-shRNA that can generate intracellular expression of small RNA molecules to achieve long-term silencing of endogenous mammalian genes. This vector is based on pcDNA 3.0, containing the human H1 promoter. This promoter is transcribed by RNA polymerase III. The original CMV promoter was deleted to prevent unexpected...