Person: ÖZBAŞ, SUNA
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ÖZBAŞ
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SUNA
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Publication Open Access A new gene therapy approach by tenascin-c genome editing induces apoptosis and cell cycle arrest in triple- negative breast cancer cells(2023-02-01) ÖZBAŞ, SUNA; Bareke H., ŞALVA E., ÖZBAŞ S.BACKGROUND/AIMS: There is a pressing need for new therapies for the most aggressive subtype of breast cancer, triple-negative breast cancer (TNBC). Tenascin-C (TN-C) codes for a tumor microenvironment-specific protein, which promotes apoptosis evasion and cell proliferation. The aim of this study was to knock down TN-C by using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system to induce cancer cell apoptosis and stunt cell proliferation, laying the grounds for a new gene therapy approach in TNBC.MATERIALS and METHODS: The human TNBC cell line, MDA-MB-231 cells were transfected by TN-C-specific CRISPR/Cas9 plasmids. TN-C messenger RNA levels were assessed by real-time polymerase chain reaction to determine the knock-down efficiency. Two days after the transfection, the percentage of apoptotic cells and the proportion of cells in cell cycle phases were compared between the treatment and the control groups using flow cytometry. The resultant change in cell proliferation due to the knock-down was determined by MTT assay. RESULTS: Transfection with the TN-C CRISPR/Cas9 plasmid reduced TN-C levels in the cells by approximately 49% relative to the scrambled -control CRISPR/Cas9 transfected cells. This TN-C downregulation increased the percentage of cells in apoptosis and induced G1-phase arrest. The combined effect of apoptosis and cell cycle arrest led to a significant decrease in the number of cancer cells in the treatment group.CONCLUSION: Our successful preliminary study of a potential TNBC gene therapy based on TN-C genome editing by the CRISPR/Cas9 system led to significant decrease in TNBC cell numbers and it justifies the testing of this system in more advanced preclinical studies.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 Chitosan-based delivery of CRISPR-Cas9 plasmid in breast cancer stem cells(2023-01-01) EKENTOK ATICI, CEYDA; ÖZBAŞ, SUNA; Canak-Ipek T., Avci-Adali M., EKENTOK ATICI C., ŞALVA E., ÖZBAŞ S.© 2023 Marmara University Press.Clustered regularly interspaced short palindromic repeat (CRISPR)-associated Cas9 nuclease system (CRISPR/Cas9) has emerged as a powerful toolbox for cancer therapy, serving as a gene fixed-point knock-out method. However, suitable gene carrier systems are urgently needed to encapsulate the CRISPR/Cas9 system and to improve the uptake into the cancer cells for anti-cancer therapy. In cancer therapy, breast cancer stem cells should be also targeted besides tumor cells. In this study, we prepared chitosan/CRISPR-Cas9/protamine nanoplexes and performed in vitro characterization. The results showed that the chitosan/protamine complex increased the zeta potential of the VEGF CRISPR/Cas9 plasmid from negative to positive. In vitro cell culture studies showed that VEGF silencing efficiency was 46.19% and 30.2% in MCF-7 and MCF-7s, respectively, after 7 days. The invasion capacity of cancer cells decreased significantly for both cell types. The results indicate that chitosan/VEGF CRISPR/Cas9 plasmid/protamine complexes can be used to reduce VEGF expression, leading to a decrease in the invasion capacity of breast cancer as well as breast cancer stem cells and providing proof of concept for more advanced studies, including in vivo studies, of this system.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.Publication Open Access Investigation of therapeutic effects in the wound healing of chitosan/pGM-CSF complexes(2022-01-01) ÖZBAŞ, SUNA; ŞALVA E., ALAN S., Karakoyun B., Cakalagaoglu F., ÖZBAŞ S., Akbuga J.Granulocyte macrophage colony-stimulating factor (GM-CSF) has been shown to promote the growth, proliferation, and migration of endothelial and keratinocyte cells. Chitosan has been widely used as a biopolymer in wound-healing studies. The aim of this study was to investigate the in vitro proliferative effects of chitosan/pGM-CSF complexes as well as the therapeutic role of the complexes in an in vivo rat wound model. The effect of complexes on cell proliferation and migration was examined. Wounds were made in Wistar-albino rats, and examined histopathologically. The cell proliferation and migration were increased weight ratio- and time-dependently in HaCaT and NIH-3T3 cell lines. Wound healing was significantly accelerated in rats treated with the complexes. These results showed that the delivery of pGM-CSF using chitosan complexes could play an accelerating role in the cell proliferation, migration, and wound-healing process.