Person: ÖZBAŞ, SUNA
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ÖZBAŞ
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SUNA
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Publication Open Access Generation of stable cell line by using chitosan as gene delivery system(SPRINGER, 2016-08) EKENTOK ATICI, CEYDA; Salva, Emine; Turan, Suna Ozbas; Ekentok, Ceyda; Akbuga, JulideEstablishing stable cell lines are useful tools to study the function of various genes and silence or induce the expression of a gene of interest. Nonviral gene transfer is generally preferred to generate stable cell lines in the manufacturing of recombinant proteins. In this study, we aimed to establish stable recombinant HEK-293 cell lines by transfection of chitosan complexes preparing with pDNA which contain LacZ and GFP genes. Chitosan which is a cationic polymer was used as gene delivery system. Stable HEK-293 cell lines were established by transfection of cells with complexes which were prepared with chitosan and pVitro-2 plasmid vector that contains neomycin drug resistance gene, beta gal and GFP genes. The transfection efficiency was shown with GFP expression in the cells using fluorescence microscopy. Beta gal protein expression in stable cells was examined by beta-galactosidase assay as enzymatically and X-gal staining method as histochemically. Full complexation was shown in the above of 1/1 ratio in the chitosan/pDNA complexes. The highest beta-galactosidase activity was obtained with transfection of chitosan complexes. Beta gal gene expression was 15.17 ng/ml in the stable cells generated by chitosan complexes. In addition, intensive blue color was observed depending on beta gal protein expression in the stable cell line with X-gal staining. We established a stable HEK-293 cell line that can be used for recombinant protein production or gene expression studies by transfecting the gene of interest.Publication Open Access In vitro gene silencing effect of chitosan/shRNA PDGF-D nanoparticles in breast cancer(MARMARA UNIV, FAC PHARMACY, 2017-10-03) EKENTOK ATICI, CEYDA; Ekentok, Ceyda; Turan, Suna Ozbas; Akbuga, JulideBreast cancer is the most common cancer worldwide in women and it is highly malignant and fatal. PDGF-D plays role in regulation of many cellular processes such as angiogenesis. PDGF-D is overexpressed in many types of cancers and promote tumor growth and metastasis. Silencing of PDGF-D gene by using shRNA with an appropriate carrier system may decrease tumor growth and metastasis. In our study, we prepared chitosan nanoparticles loaded with five different shRNA plasmids targeting different exons of PDGF-D gene. Then, nanoparticles were characterized in vitro and transfection efficiency of these nanoparticles were investigated in breast cancer cell lines (MCF7, MDA-MB-231 and MDA-MB-435). The effects of single and multiple shRNA sequences, molecular weight of chitosan (150 kDa and 400 kDa) and the amount of shRNA (100 and 500 mu g) on the characterization and transfection efficiencies of nanoparticles have been studied. Size of nanoparticles changed between 200-400 nm and approximately 95-100% encapsulation efficiency were obtained. Release of shRNA changed with the molecular weight of chitosan. It was obtained that formulation containing shRNA plasmid targeting PDGF-D exon 6 (NP1) has the highest silencing efficiency in MDA-MB-231 cell line. It was also evaluated that chitosan can be a suitable gene delivery system for shRNA targeting PDGF-D.Publication Metadata only Investigation of antineoplastic activity of achillea nobilis subsp. Neilreichii plant in experimental breast cancer(2019-07-03) YAVUZ, AYŞE NUR; EKENTOK ATICI, CEYDA; TAŞKIN, TURGUT; ÖZBAŞ, SUNA; KABASAKAL, LEVENT; Sehlan S. S., Yavuz A. N., Ekentok Atıcı C., Taşkın T., Alan S., Özbaş S., Kabasakal L.Publication Metadata only Mammaglobin gen ekspresyonunun kitozan/siRNA nanopleksleri ile baskılanması ve in vitro karakterizasyonu(2018-11-16) EKENTOK ATICI, CEYDA; ÖZBAŞ, SUNA; Özkavak Ş. B., Ekentok Atıcı C., Şalva E., Özbaş S.Publication Metadata only The effects of some traditional medical plants and beta amyloid protein in cell viability(2019-07-03) YAVUZ, AYŞE NUR; EKENTOK ATICI, CEYDA; TAŞKIN, TURGUT; ÖZBAŞ, SUNA; KABASAKAL, LEVENT; Saleh Al-Rabeei M. A., Yavuz A. N., Ekentok Atıcı C., Taşkın T., Özbaş S., Kabasakal L.Publication Metadata only Kitozan/mammaglobin shRNA nanoplekslerinin hazırlanması ve in vitro karakterizasyonu(2018-11-16) ÖZBAŞ, SUNA; EKENTOK ATICI, CEYDA; Özkan A., Ekentok Atıcı C., Şalva E., Özbaş S.Meme kanseri kadınlar arasında dünyada en sık görülen malign tümörlerden biridir. İnsan mammaglobin (hMAM), genellikle meme ve meme kanseri hücrelerinde eksprese edilen bir glikoproteindir. Meme spesifik hMAM’ın RNA interferans (RNAi) teknolojisi ile short hairpin RNA (shRNA) kullanılarak baskılanması önemli bir gen tedavi stratejisidir. Nükleaz degredasyonundan korunma ve hücreye giriş etkinliğini arttırmak için viral olmayan gen taşıyıcı sistem olarak kitozan; biyobozunur olması, toksik olmayışı ve katyonik yapısı sebebiyle sıklıkla tercih edilmektedir. Bu çalışmada da, hMAM genini baskılama amacıyla kitozan/mammaglobin shRNA nanopleksleri hazırlanmıştır. Bu amaçla, transformasyon sonrası [1] izole edilen shRNA’nın spektrofotometrik ve elektroforetik kontrolleri yapılmıştır [2]. Elektrostatik etkileşim yöntemiyle farklı oranlarda (0.5/1, 1/1, 2/1, 3/1, 4/1, 5/1; a/a) kitozan/shRNA nanopleksleri hazırlanarak partikül boyutu, zeta potansiyel, jel elektroforez ve serum stabilitesi analizleri yapılmıştır. Elektroforez sonuçlarına göre tam kompleks oluşumu gözlenmiş ve orana bağlı olarak nanopleks boyutlarının 153.2 ile 436.3 nm arasında, zeta potansiyel değerlerinin de +0.6 ile +31.9 mV arasında olduğu belirlenmiştir. Serum stabilitesi sonuçlarına göre hazırlanan nanoplekslerin shRNA’yı degredasyondan koruduğu görülmüştür. Elde edilen sonuçlar ışığında, hazırlanan formülasyonların meme kanseri tedavisindeki etkinliğinin araştırılması için ileri in vitro ve in vivo çalışmalar planlanmaktadır.