Effects of chitosan nanoparticles with long synthetic siRNAs targeting VEGF in triple-negative breast cancer cells

Vascular endothelial growth factor (VEGF) is an essential angiogenic factor in breast cancer development and metastasis. Small interfering RNAs (siRNAs) can specifically silence genes via the RNA interference pathway, therefore were investigated as cancer therapeutics. In this study, we investigated the effects of siRNAs longer than 30 base pairs (bp) loaded into chitosan nanoparticles in triple-negative breast cancer cells, compared with conventional siRNAs. 35 bp long synthetic siRNAs inhibited VEGF gene expression by 51.2% and increased apoptosis level by 1.75-fold in MDA-MB-231 cell lines. Furthermore, blank and siRNA-loaded chitosan nanoparticles induced expression of IFN-γ in breast cancer cells. These results suggest that long synthetic siRNAs can be as effective as conventional siRNAs, when introduced into cells with chitosan nanoparticles.Marmara Universit

Uzun siRNA içeren kitozan nanopartikülleri ile meme kanseri hücrelerinde vasküler endotelyal büyüme faktörü geninin susturulması

ÖZETAmaç: VEGF, meme kanserinin gelişiminde ve metastazında etkili olan önemli bir anjiyogenik faktördür. RNAi mekanizmasını başlatan siRNA, hedeflenen genin transkripsiyon sonrası diziye özgü bir şekilde inhibe edilmesini sağlayarak kanser gibi hastalıkların hedeflenmesi için gen tedavisinde önemli bir araç haline gelmiştir. Bu çalışmada, farklı meme kanseri hücre hatlarında uzun (35 bp) siRNA içeren kitozan nanopartikülleri ile VEGF ekspresyonunun baskılanması ve kısa (21 bp) siRNA’nın gen inhibisyonu etkisiyle karşılaştırılması amaçlanmıştır.Gereç ve yöntem: 21 bp ve 35 bp siRNA içeren kitozan nanopartiküller TPP kullanılarak iyonik jelasyon yöntemiyle hazırlandı. Elde edilen nanopartiküllerin partikül boyutu, zeta potansiyeli, stabilite ve morfolojik özellikleri incelendi. Hazırlanan nanopartiküllerin MCF-7, MDA-MB-231, MDA-MB-435 ve SKBR-3 hücre hatlarında gen inhibisyonu üzerine etkileri incelendi. siRNA içeren kitozan nanopartiküllerin transfeksiyon sonrası in vitro interferon cevabı ve apoptoz üzerine etkileri araştırıldı.Bulgular: Uzun siRNA ile 324.3 ± 17.9 nm boyutunda ve 10.0 ± 3.0 mV yüzey yüküne sahip kitozan nanopartiküller elde edildi ve dört farklı meme kanseri hücre hattında VEGF gen inhibisyonu (% 45.4- 51.5) sağlandı. Meme kanseri hücre hatlarında siRNA içeren kitozan nanopartiküllerin in vitro IFN-γ ekspresyonuna neden olduğu ve apoptozu indüklediği gösterildi. Sonuç: Uzun ve kısa siRNA’nın kitozan nanopartiküller ile hücrelere alınarak etkin bir şekilde VEGF ekspresyonunu azalttığı ve her ikisi için de yakın sonuçlar elde edildiği gösterilmiştir.ABSTRACTAim: VEGF is an important angiogenic factor in development and metastasis of breast cancer. siRNA triggering the RNAi mechanism, has become an important tool in gene therapy for the targeting of diseases such as cancer, allowing to post-transcriptional inhibition of the target gene in a sequence specific manner. In this study, it was aimed to supress of VEGF expression with chitosan nanoparticles containing long (35 bp) siRNA in different breast cancer cell lines and to compare gene inhibition activity of short (21 bp) siRNA.Material and Method: Chitosan nanoparticles containing 21 bp and 35 bp siRNA were prepared by ionic gelation method using TPP. The particle size, zeta potential, stability and morphological properties of the obtained nanoparticles were investigated. The gene inhibition activities of the prepared nanoparticles were examined on MCF-7, MDA-MB-231, MDA-MB-435 and SKBR-3 cell lines. After transfection with siRNA-containing chitosan nanoparticles, their effects on in vitro interferon response and apoptosis were investigated.Results: Chitosan nanoparticles with 324.3 ± 17.9 nm size and 10.0 ± 3.0 mV surface charge were obtained with long siRNA and VEGF gene inhibition (45.4-51.5%) was achieved in four different breast cancer cell lines. siRNA-containing chitosan nanoparticles were shown to induce in vitro IFN-γ expression and apoptosis in the breast cancer cell lines.Conclusion: It has been shown that long and short siRNAs reduce VEGF expression effectively by delivering them into the cells with chitosan nanoparticles and close results were obtained for the both of them

Uzun siRNA içeren kitozan nanopartikülleri ile meme kanseri hücrelerinde vasküler endotelyal büyüme faktörü geninin susturulması

Amaç: VEGF, meme kanserinin gelişiminde ve metastazında etkili olan önemli bir anjiyogenik faktördür. RNAi mekanizmasını başlatan siRNA, hedeflenen genin transkripsiyon sonrası diziye özgü bir şekilde inhibe edilmesini sağlayarak kanser gibi hastalıkların hedeflenmesi için gen tedavisinde önemli bir araç haline gelmiştir. Bu çalışmada, farklı meme kanseri hücre hatlarında uzun (35 bp) siRNA içeren kitozan nanopartikülleri ile VEGF ekspresyonunun baskılanması ve kısa (21 bp) siRNA’nın gen inhibisyonu etkisiyle karşılaştırılması amaçlanmıştır. Gereç ve yöntem: 21 bp ve 35 bp siRNA içeren kitozan nanopartiküller TPP kullanılarak iyonik jelasyon yöntemiyle hazırlandı. Elde edilen nanopartiküllerin partikül boyutu, zeta potansiyeli, stabilite ve morfolojik özellikleri incelendi. Hazırlanan nanopartiküllerin MCF-7, MDA-MB-231, MDA-MB-435 ve SKBR-3 hücre hatlarında gen inhibisyonu üzerine etkileri incelendi. siRNA içeren kitozan nanopartiküllerin transfeksiyon sonrası in vitro interferon cevabı ve apoptoz üzerine etkileri araştırıldı. Bulgular: Uzun siRNA ile 324.3 ± 17.9 nm boyutunda ve 10.0 ± 3.0 mV yüzey yüküne sahip kitozan nanopartiküller elde edildi ve dört farklı meme kanseri hücre hattında VEGF gen inhibisyonu (% 45.4- 51.5) sağlandı. Meme kanseri hücre hatlarında siRNA içeren kitozan nanopartiküllerin in vitro IFN-γ ekspresyonuna neden olduğu ve apoptozu indüklediği gösterildi. Sonuç: Uzun ve kısa siRNA’nın kitozan nanopartiküller ile hücrelere alınarak etkin bir şekilde VEGF ekspresyonunu azalttığı ve her ikisi için de yakın sonuçlar elde edildiği gösterilmiştir. ABSTRACT Aim: VEGF is an important angiogenic factor in development and metastasis of breast cancer. siRNA triggering the RNAi mechanism, has become an important tool in gene therapy for the targeting of diseases such as cancer, allowing to post-transcriptional inhibition of the target gene in a sequence specific manner. In this study, it was aimed to supress of VEGF expression with chitosan nanoparticles containing long (35 bp) siRNA in different breast cancer cell lines and to compare gene inhibition activity of short (21 bp) siRNA. Material and Method: Chitosan nanoparticles containing 21 bp and 35 bp siRNA were prepared by ionic gelation method using TPP. The particle size, zeta potential, stability and morphological properties of the obtained nanoparticles were investigated. The gene inhibition activities of the prepared nanoparticles were examined on MCF-7, MDA-MB-231, MDA-MB-435 and SKBR-3 cell lines. After transfection with siRNA-containing chitosan nanoparticles, their effects on in vitro interferon response and apoptosis were investigated. Results: Chitosan nanoparticles with 324.3 ± 17.9 nm size and 10.0 ± 3.0 mV surface charge were obtained with long siRNA and VEGF gene inhibition (45.4-51.5%) was achieved in four different breast cancer cell lines. siRNA-containing chitosan nanoparticles were shown to induce in vitro IFN-γ expression and apoptosis in the breast cancer cell lines. Conclusion: It has been shown that long and short siRNAs reduce VEGF expression effectively by delivering them into the cells with chitosan nanoparticles and close results were obtained for the both of them

Effects of Chitosan Nanoparticles with Long Synthetic siRNAs Targeting VEGF in Triple-Negative Breast Cancer Cells

Vascular endothelial growth factor (VEGF) is an essential angiogenic factor in breast cancer development and metastasis. Small interfering RNAs (siRNAs) can specifically silence genes via the RNA interference pathway, therefore were investigated as cancer therapeutics. In this study, we investigated the effects of siRNAs longer than 30 base pairs (bp) loaded into chitosan nanoparticles in triple-negative breast cancer cells, compared with conventional siRNAs. 35 bp long synthetic siRNAs inhibited VEGF gene expression by 51.2% and increased apoptosis level by 1.75-fold in MDA-MB-231 cell lines. Furthermore, blank and siRNA-loaded chitosan nanoparticles induced expression of IFN-γ in breast cancer cells. These results suggest that long synthetic siRNAs can be as effective as conventional siRNAs, when introduced into cells with chitosan nanoparticles

Preparation and in vitro characterization of laminarin based hydrogels

Hydrogels are one of the most effective pharmaceutical forms because of several advantages in the treatment of burn and wound headings. Laminarin is a storage polysaccharide derived from brown algae and member of the 1-3-beta-D-glucan family. It is a heparin-like molecule and plays role in hemoslasis, regulation of cell growth and regeneration. Also, indirectly functional in neovascularizalion. In addition, accelerates the formation of fibroblast activity and re-epithelization. Aim of this study is prepare laminarin hydrogels and evaluate the ill vitro characteristics of hydrogels for use in various wounds. For this purpose, different hydrogel formulations prepared and examined, and evaluated by water absorption Ca racily, viscosity, rheological and mechanical properties and bioadhesion. II was observed that hydrogels prepared with laminarin are an ideal wound dressing as expected