PRODUCTION AND CHARACTERIZATION OF siRNA LOADED MAGNETIC NANOPARTICLES  TO BE USED IN CANCER TREATMENT

SELIMOVIC, AMINA

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2017

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SELIMOVIC, AMINA

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ABSTRACT PRODUCTION AND CHARACTERIZATION OF siRNA LOADED MAGNETIC NANOPARTICLES TO BE USED IN CANCER TREATMENT AMINA SELIMOVIC Master of Science, Bioengineering Supervisor: Prof. Dr. Emir Baki DENKBAŞ June 2017, 73 pages Cancer is one of the leading causes of death in economically developed and developing countries and factors such as smoking, physical inactivity and nutritional habits are increasing the possibility of its development. After decades of extensive research of cancer, today it can be described as a genetic disease of the somatic cell. Genetic changes such as rearrangements of chromosomes (deletion, translocation, insertion), point mutation, gene amplification are affecting kinase inhibitors, growth factors, receptors, a cascade of transcription factors and signal transduction members are leading to an impaired balance of cell proliferation and changes in the function of genes that induce apoptosis. These actions are leading to the abnormal growth of the cells called neoplasia. An approach based on nanotechnology provides a great promise in developing strategies for cancer treatment by helping to improve the safety and efficacy of therapeutic delivery vehicles. Powerful investigational tools with great potential in therapeutics-RNA interference (RNAi) is known as a highly efficient regulatory process in which short double-stranded RNAs are giving rise to sequence-specific posttranscriptional gene silencing. With time it has been proven that specific protein expression can be inhibited. Small interfering RNA (siRNA) as part of RNA interference process has been extensively studied to treat various genetic diseases, cardiovascular diseases and various cancers. However, due to its polyanionic nature siRNAs cannot cross the cellular membrane that is why it needs a carrier to prevent enzymatic degradation and to take siRNA to the specific target inside the cell. Properties such as safety, effectiveness, ease of manufacturing and production are quite important to consider when selecting a proper carrier for siRNA. In this work iron oxide nanoparticles are coated with natural biopolymer gelatin, loaded with mTOR siRNA targeting specific oncogene with the aim to deliver gene silencing complex to colon cancer cells inducing therapeutic effect. For this iron oxide and gelatin coated iron oxide nanoparticles were synthesized, optimized and characterized. Morphological characterization was done using SEM (Scanning Electron Microscopy). Size and surface charge of produced nanoparticles was revealed by Zeta-Sizer (3000 HSA, Malvern, England). To determine the chemical structures of the nanoparticles, molecular bond characterization had been performed using Fourier Transform Infrared Spectroscopy (FTIR) (Nicolet iS10, USA). VSM (vibrating sample magnetometer) and ESR (electro spin resonance) are used to analyse magnetic properties of the prepared particles. siRNA was loaded to gelatin coated iron oxide nanoparticles and its binding efficiency (%) was examined. siRNA loaded nanoparticles were transfected to colon cancer cell line CaCo-2 and mouse fibroblast cell line L929. Cell cytotoxicity test, MTT was performed using different concentrations of siRNA and under different incubation time MTT assay showed that toxic effect in both cell lines was significantly higher when siRNA loaded gelatin coated IONs were used. Also, according to the results obtained, synthesized gelatin coated IONs showed similar anticancer activity as HiperFect which is commercial siRNA carrier. This work showed that gelatin coated iron oxide nanoparticles as cheap and easily synthesized carrier are promissing tool for siRNAs delivery. Keywords: nanooncology, iron oxide nanoparticles, gelatin, siRNA

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http://hdl.handle.net/11655/3769

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Biyomühendislik Bölümü Tez Koleksiyonu [98]

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