Applications of the multifunctional magnetic nanoparticles for development of molecular therapies for breast cancer

Download
2015
Aşık, Elif
The understanding of how magnetic nanoparticles (MNPs) interact with living system is one of the prerequisite pieces of information needed to be obtained before any further development for desired biomedical applications. In this study, Cobalt Ferrite magnetic nanoparticles (CoFe-MNPs) in their naked and silica-coated forms were characterized. In vitro cell culture for their likely cytotoxicity and genotoxicity potential were examined. The apoptosis, lipid peroxidation, ROS formation and oxidative stress related gene expression levels of some drug metabolizing enzymes in human cancer (MDA-MB- 231, MCF-7) and non-cancer (MCF-10A) breast cell lines were analyzed in response to MNPs treatment. Our results revealed that, uptake of the highest amounts of CoFe-MNPs was observed in metastatic cells and the uptake of the silica coated CoFe-MNPs were higher than the naked ones in all cells. Naked CoFe-MNPs represented higher levels of cytotoxicity, genotoxicity and ROS generation compared to silica coated CoFe-MNPs. Silica coated CoFe-MNPs were functionalized with COOH groups for further modifications. 2-amino-2-deoxy-glucose (2DG), which is a potential targeting molecule for cancer treatment, was conjugated on silica coated CoFe-MNPs surface through - vi COOH groups. Internalization and accumulation of both -COOH modified (COOHMNPs) and 2DG conjugated (2DG-MNPs) were studied in MDA-MB-231 and MCF-7 cancer and MCF-10A non-cancer breast cells by transmission electron microscopy (TEM), Prussian blue staining and Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). According to our results, it was apparent that 2DG-MNPs were internalized more efficiently than COOH-MNPs under same conditions, in all cell types studied. Moreover, the highest amount of uptake was observed in MDA-MB-231 cells, which is followed by MCF-7 and normal MCF-10A for both MNPs. The apoptotic effects of 2DG-MNPs was further evaluated, and it was found that apoptosis was not induced at low concentration of 2DG-MNPs, regardless of the cell types, whereas dramatic cell death was observed at higher concentrations. In addition, the gene expression levels of some drug metabolizing enzymes, two Phase I (CYP1A1, CYP1B1) and two Phase II (GSTM3, GSTZ1) were also seen to increase at high concentration of 2DG-MNPs, whereas at low concentration no induction was observed. Eukaryotic elongation factor-2 kinase (eEF2K) is gaining potential as a prognostic marker and therapeutic target to treat breast cancer. Hence, eEF2K siRNA was decorated on silica coated CoFe-MNPs through -COOH groups and the expression and role of this gene investigated in the BRCA1 mutated breast cancer cells (MDA-MB-436, HCC- 1937). According to our results, the silencing of eEF2K using siRNA decorated MNPs inhibited colony formation; proliferation, migration and invasion of BRCA1 mutated cells. Furthermore, the down modulation of eEF2K was investigated in an orthotropic model of BRCA1 mutated breast cancer in nude mice in vivo by through systematically administration of eEF2K siRNA decorated MNPs. In vivo silencing of eEF2K lead to inhibition of molecules and pathways that are involved in migration/invasion (Src/FAK/paxillin), angiogenesis (VEGF), proliferation (c-Myc), cell cycle (CyclinD1), survival/drug resistance (PI3K/Akt) translational regulation (4EBP1). Taken together, our data suggest, for the first time, which eEF2K is associated with tumorigenesis and progression of BRCA1 mutated breast cancer and may be a novel potential therapeutic target in this cancer

Suggestions

Investigation of Multi-task Deep Neural Networks in Automated Protein Function Prediction
Rifaioğlu, Ahmet Süreyya; Martin, Maria Jesus; Atalay, Rengül; Atalay, Mehmet Volkan; Doğan, Tunca (2017-07-20)
Functional annotation of proteins is a crucial research field for understanding molecular mechanisms of living-beings and for biomedical purposes (e.g. identification of disease-causing functional changes in genes and for discovering novel drugs). Several Gene Ontology (GO) based protein function prediction methods have been proposed in the last decade to annotate proteins. However, considering the prediction performances of the proposed methods, it can be stated that there is still room for significant imp...
Assessment of changes in the dielectric properties of multidrug resistant cancer cells by electrorotation technique /
Bahrieh, Garsha; Külah, Haluk; Department of Electrical and Electronics Engineering (2014)
With recent advances in microfabrication technologies different methodologies are employed to study the dielectric properties of the biological systems. These include electrorotation (ER), dielectrophoresis, microelectrical impedance spectroscopy, and impedance flow cytometry. Among them, ER is utilized as a high accuracy approach in determining the membrane and interior dielectric properties of single cells. In addition, in cancer therapy, cancer cells are developing resistance toward the chemotherapeutic ...
Data Acquisition System for Harmonic Motion Microwave Doppler Imaging
Tafreshi, Azadeh Kamali; Karadas, Muersel; Top, Can Baris; Gençer, Nevzat Güneri (2014-08-30)
Harmonic Motion Microwave Doppler Imaging (HMMDI) is a hybrid method proposed for breast tumor detection, which images the coupled dielectric and elastic properties of the tissue. In this paper, the performance of a data acquisition system for HMMDI method is evaluated on breast phantom materials. A breast fat phantom including fibro-glandular and tumor phantom regions is produced. The phantom is excited using a focused ultrasound probe and a microwave transmitter. The received microwave signal level is mea...
Integration of a Glutamate Sensitive Genetically Encoded Sensor Protein into Photocrosslinkable Hydrogel Optrodes
Kahyaoğlu, Leyla Nesrin (2016-01-01)
Immobilization into 3D matrices stabilizes proteins in comparison to flat planar surfaces and facilitates the study of the biomolecular interactions as well as integration into optrodes for cell physiology. Photocrosslinkable hydrogels have received significant attention in recent years as they provide not only a highly hydrophilic 3D environment to promote protein stabilization and its interactions with analyte molecules, but enable optically addressable patterning for spatial control of protein localizati...
Investigation of SNX3 in colon cancer
Cingöz, Harun; Erson Bensan, Ayşe Elif; Department of Biology (2019)
Sorting Nexin 3 (SNX3) is part of the retromer complex that recycles cargo receptors back to plasma membrane or to Trans Golgi Network. WNT ligand carrier protein Wntless (WLS) is a known SNX3 cargo protein. Our earlier data suggested over expression of SNX3 in colon cancer cells. Considering its importance in receptor recycling, we hypothesized SNX3 to be a potential modulator of cancer related receptors. To begin understanding the role of SNX3, we developed RNAi models of SNX3 in SW480 colon cancer cells ...
Citation Formats
E. Aşık, “Applications of the multifunctional magnetic nanoparticles for development of molecular therapies for breast cancer,” Ph.D. - Doctoral Program, Middle East Technical University, 2015.