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- Title
TARGETING ANGIOGENESIS WITH siRNA IN CANCER.
- Authors
Pileczki, Valentina; Braicu, Cornelia; Balacescu, O.; Dragos, N.; Berindan-Neagoe, Ioana
- Abstract
Synthetic siRNA is used in mammalian cells for gene silencing. siRNA-induced RNAi is a key strategy for investigating gene function, a method that reduces the expression of individual genes in order to establish a link between gene identity and gene function. In our experiments we used siRNA VEGF to inhibit the main factor of angiogenesis that is considered to play an important role in tumor growth and metastasis. In this study a VEGF small interfering RNA (siRNA) was synthesized and through reverse-transfection technology was introduced in hepatocellular carcinoma cell line HepG2, using siPORT™ NeoFX Transfection Agent. VEGF expression was analysed by real time PCR, and the results were confirmed at protein levels by ELISA assay. MTT assay was used to detect cell viability. Apoptosis was analyzed using DAPI staining. The evaluation of real-time PCR gene expression showed a downregulation of VEGF at mRNA level in transfected cells compared to control group. These results were confirmed also at a protein level. VEGF-siRNA cell growth inhibition assessed by the MTT assay, showed that the transfection agent had no cytotoxic effect at 24 and 48 hours. An increased level of apoptotic processes was observed especially at 48 hours. MTT analysis shows that the transfection agent has no cytotoxic effect; the low cellular proliferation rate may be caused by the inhibition of VEGF gene and protein expression leading to the activation of apoptotic pathways.
- Subjects
SMALL interfering RNA; GENE silencing; CANCER; METASTASIS; CELL culture; CELL lines; VASCULAR endothelial growth factors
- Publication
Annals of the Romanian Society for Cell Biology, 2011, Vol 16, Issue 2, p20
- ISSN
2067-3019
- Publication type
Article