Publication: Hepatosellüler karsinoma hücre hatlarında mTOR (Mammalian Target of Rapamycin) gen ekspresyonunun plazmid bazlı mikroRNA-7/ kitozan kompleksleri ile baskılanmasına ilişkin in vitro çalışmalar
Abstract
Amaç: Tez çalışmasında amaç; hepatosellüler karsinomada ekspresyonu downregüle olan miR-7'nin özellikleri incelerek katyonik bir biyopolimer olan kitozanla hazırlanan kompleks formlarının Hep 3B ve Hep G2 hepatosellüler karsinoma hücre hatlarındaki etkinliği ve hepatosellüler karsinoma tedavisindeki kullanılabilirliği in vitro olarak araştırmaktır.Gereç ve Yöntem: Hep 3B ve Hep G2 hepatosellüler karsinoma hücrelerinde mTOR geninin ekspresyonunu baskılayabilecek miR-7'nin etkin bir şekilde aktarılabilmesi için doğal kitozan polimerleri ile kompleksler hazırlanmıştır. Hazırlanan komplekslerin partikül boyutu ve yüzey yükü ölçülerek fizikokimyasal karakterizasyonları yapılmıştır. Taşıyıcı sistemlerin ise serum stabilitesi, sitotoksisitesi, transfeksiyon etkinliği, hepatosellüler karsinoma hücrelerinin migrasyonu üzerine etkisi değerlendirilmiştir.Bulgular: Farklı oranlarda kitozan kullanılarak hazırlanan kompleksler ile Hep 3B ve Hep G2 hepatosellüler karsinoma hücresine transfeksiyonu sağlanmıştır. Kitozan ile 1/ 1 ve 2/ 1 oranında hazırlanmış miR-7 tam kompleks oluşumu sağlamıştır.Sonuçlar: Çalışma sonucunda miR-7'nin kitozan kompleksleri ile hücrelere stabil olarak taşındığı, hücreye internalize olarak kanser hücrelerinde bozulan miRNA regülasyonunun tedavi edilmesi ile kanser hücrelerinin invazivliğini azalttığı gösterilmiştir.
Objective: The aim of this study is to investigate the properties of miR-7 which is downregulated expression in hepatocellular carcinoma and to investigate its efficacy in Hep 3B and Hep G2 hepatocellular carcinoma cell lines and its usefulness in hepatocellular carcinoma treatment in vitro.Material and Method: Complexes with natural chitosan polymers were prepared to efficiently transfer miR-7, which can suppress the expression of mTOR gene in Hep 3B and Hep G2 hepatocellular carcinoma cells. The particle size and surface charge of the prepared complexes were measured and physicochemical characterizations were made. The effects of carrier systems on serum stability, cytotoxicity, transfection efficiency, and migration of hepatocellular carcinoma cells were evaluated.Result: Transfection of Hep 3B and Hep G2 hepatocellular carcinoma cells was achieved with complexes prepared using chitosan in different ratios. MiR-7 prepared with chitosan in a ratio of 1/ 1 and 2/ 1 provided complete complex formation.Conclusion: As a result of this study, it was shown that miR-7 was transported to cells with chitosan complexes and decreased the invasiveness of cancer cells by treatment of miRNA regulation in cancer cells which were internalized to the cell.
Objective: The aim of this study is to investigate the properties of miR-7 which is downregulated expression in hepatocellular carcinoma and to investigate its efficacy in Hep 3B and Hep G2 hepatocellular carcinoma cell lines and its usefulness in hepatocellular carcinoma treatment in vitro.Material and Method: Complexes with natural chitosan polymers were prepared to efficiently transfer miR-7, which can suppress the expression of mTOR gene in Hep 3B and Hep G2 hepatocellular carcinoma cells. The particle size and surface charge of the prepared complexes were measured and physicochemical characterizations were made. The effects of carrier systems on serum stability, cytotoxicity, transfection efficiency, and migration of hepatocellular carcinoma cells were evaluated.Result: Transfection of Hep 3B and Hep G2 hepatocellular carcinoma cells was achieved with complexes prepared using chitosan in different ratios. MiR-7 prepared with chitosan in a ratio of 1/ 1 and 2/ 1 provided complete complex formation.Conclusion: As a result of this study, it was shown that miR-7 was transported to cells with chitosan complexes and decreased the invasiveness of cancer cells by treatment of miRNA regulation in cancer cells which were internalized to the cell.