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https://olympias.lib.uoi.gr/jspui/handle/123456789/33348| Title: | MiR-9, an IKKα-regulated miRNA in lung cancer |
| Institution and School/Department of submitter: | Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας |
| Keywords: | lung cancer,microRNA,miR-9-5p,IKKα |
| URI: | https://olympias.lib.uoi.gr/jspui/handle/123456789/33348 http://dx.doi.org/10.26268/heal.uoi.13064 |
| Table of contents: | Contents Acknowledgements 2 Summary 5 Περίληψη 7 1. Introduction 10 1.1. Lung cancer and its molecular mechanisms 10 1.1.1. Genetic changes in lung cancer 10 1.1.2. Epigenetic changes in lung cancer 12 1.2. The role miRNAs in cancer regulation 13 1.2.1. MicroRNA biogenesis 14 1.2.2. MiRNA mechanism of action 15 1.2.3. MiRNAs in cancer 16 1.2.4. MiRNAs in lung cancer 17 1.2.5. MiR-9-5p in cancer 18 1.2.6. MiR-9-5p in lung cancer 21 1.2.7. MiRNAs as therapy targets 22 1.3. The NF-κB pathway in cancer 23 1.3.1 NF-κB signalling in lung cancer 25 Aims 28 2. Materials and Methods 29 2.1. Bacterial cell methods 29 2.1.1. Preparation of L-broth (Luria-Bertani; LB) and L-agar plates 29 2.1.2. Preparation of competent bacteria 29 2.1.3. Bacterial Transformation with plasmid DNA 30 2.1.4. Isolation of Plasmid DNA on a Small Scale (Mini Prep) 30 2.1.5. Isolation of Plasmid DNA on a Large Scale (Midi Prep) 30 2.2. Tissue culture methods 31 2.2.1. Cell culture 31 2.2.2. Cryopreservation of cell lines 31 2.2.3. Stable cell line generation 31 2.2.4. Cell Proliferation Assay 34 2.2.5. Flow cytometry (FACs) for cell cycle analysis 34 2.2.6. Matrigel Invasion Assay 35 2.2.7. Tumour xenografts 35 2.3. Molecular and Biochemical methods 36 2.3.1. RNA Isolation 36 2.3.2. Total RNA extraction from mouse xenografts 36 2.3.2. Nanostring Analysis – miRNome profiling 36 2.3.3. cDNA synthesis 36 2.3.4. Quantitative Polymerase Chain Reaction (qPCR) 37 2.3.5. Target Prediction Algorithms 37 2.3.6. Total Protein Extraction 37 2.3.7. Total protein Isolation from mouse xenografts 38 2.3.8. Western Blotting 38 2.4. Statistical Analysis 39 3. Results 40 3.1. Nanostring analysis of IKKαKD cell lines 40 3.2. Generation of miR-9-5p overexpression cell lines 42 3.3. miR-9-5p effect on cell proliferation, cell cycle and invasion in vitro 43 3.4. Predicted targets of miR-9-5p 45 3.5. miR-9-5p targets E-Cadherin in LUAD 47 3.6. miR-9-5p overexpression promotes tumour formation in vivo 49 4. Discussion 52 4.1. Introduction 52 4.2. IKKα loss induces miR-9-5p expression 53 4.3. MiR-9-5p targets in NSCLC 53 4.3.1. EMT and cancer 54 4.3.2. Wnt signalling pathway 55 4.4. miR-9-5p targets CDH1 in lung cancer 58 4.5. miR-9-5p promotes tumour growth in vivo 59 4.6. Conclusions 60 References 61 |
| Appears in Collections: | Διατριβές Μεταπτυχιακής Έρευνας (Masters) - ΙΑΤ |
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| File | Description | Size | Format | |
|---|---|---|---|---|
| Μ.Ε. Μπέστα Σιμώνη (2023).pdf | 2.12 MB | Adobe PDF | View/Open |
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