Emine Şalva, Naziye Özkan, Levent Kabasakal, Suna Özbaş Turan, Jülide Akbuğa

siRNA’nın biyodağılımına kitozan komplekslerinin etkisi

Amaç: RNAi kanser dahil olmak üzere birçok hastalığın moleküler mekanizmasının analizinde ve gen susturulmasında hücresel proseslerin kontrolü için önemli bir araçtır. VEGF sinyali meme kanserinde siRNA taşınmasında önemli bir hedeftir. siRNA farklı hastalıklar için potansiyel bir ajan olmasına rağmen, siRNA’nın intrasellüler taşınması, terapötik olarak aktif bir moleküle dönüşmesindeki önemli engellerden biridir. Bugüne kadar birçok transfeksiyon yöntemi ve taşıyıcı sistem geliştirilmiştir. Bunlar arasında kitozan, biyouyumlu, biyoparçalanabilir olması, toksik ve immunojenik olmaması gibi özellikleri nedeniyle önemli bir gen taşıyıcısıdır. Bu çalışmanın amacı, meme kanserinde kitozan/VEGF-siRNA komplekslerinin tümör lokalizasyonunu ve biyodağılımını araştırmaktır.Yöntem: Çalışmamızda meme tümörü taşıyan sıçanlara serbest FITC-işaretli siVEGF (40 µg/sıçan) ve kitozan/ FITC-işaretli siVEGF (40 µg/sıçan) kompleksleri intravenöz olarak enjekte edildi.Bulgular: Kitozan/siVEGF komplekslerinin beyin ve kalbe biyodağılımı, serbest siVEGF ile hemen hemen benzerken, dalak, karaciğer, akciğer ve kasta biraz daha düşük ve böbrekte ise biraz daha yüksektir. Meme tümör dokusunda, kompleksler enjeksiyon sonrası 15 dakikada tümörde lokalize iken, serbest FITC-siVEGF tümör dokusunda lokalize değildir.Sonuç: Bu ön çalışmada, biz biyodağılım için VEGF siRNA taşıyıcı sistem olarak kitozanın umut verici olduğunu gösterdik. Anahtar Kelimeler: siRNA, VEGF, kitozan, biyodağılım, meme kanseri

Anahtar Kelimeler:

siRNA, VEGF, kitozan, biyodağılım, meme kanseri

The effect of chitosan complexes on biodistribution of siRNA

Objective: RNAi is a powerful tool for controlling cellular processes in the gene silencing and in the analysis of molecular mechanisms for many diseases including cancer. VEGF signaling is a potential therapeutic target for siRNA delivery in breast cancer. Although siRNA can be potential therapeutic agent for various diseases, intracellular delivery of siRNA is one of the major hurdles to turn siRNA into therapeutically active molecules. To date, numerous transfection methods or delivery systems have been developed. Among them, chitosan is potential gene carrier due to its characteristics such as biodegradability, biocompatibility, non immunogenic and toxicity. The purpose of this study was to investigate the biodistribution and tumor localization of chitosan/ VEGF siRNA complexes in breast cancer model of rat.Method: In our study, we intravenously injected FITC labeled naked siRNA-VEGF (40 µg/rat) and chitosan/FITC labeled siRNAVEGF complexes (40 µg/rat) to breast tumor-bearing rats.Results: While the biodistribution of chitosan/siVEGF complexes to the brain and heart appeared almost similar to that observed for naked siVEGF, the accumulation was slightly lower in the spleen, liver, lungs, muscle and higher in the kidney. In the breast tumor tissue, chitosan/FITC-labeled VEGF siRNA complexes were localized in the tumor 15 min post-injection but naked FITCsiVEGF did not localize in tumor tissue.Conclusion: In this preliminary study, we revealed the promising potential of chitosan as VEGF siRNA delivery system for biodistribution.

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