Functional genomics of ORP2 or protrudin knockdown in human umbilical vein endothelial cells

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http://urn.fi/URN:NBN:fi:hulib-201910303803
Title: Functional genomics of ORP2 or protrudin knockdown in human umbilical vein endothelial cells
Alternative title: ORP2 ja protrudiini hiljennyksen funktionaalinen genetiikka, ihmisen napanuoran suonen soluissa
Author: Taskinen, Juuso
Other contributor: Helsingin yliopisto, Bio- ja ympäristötieteellinen tiedekunta
University of Helsinki, Faculty of Biological and Environmental Sciences
Helsingfors universitet, Bio- och miljövetenskapliga fakulteten
Publisher: Helsingin yliopisto
Date: 2019
Language: eng
URI: http://urn.fi/URN:NBN:fi:hulib-201910303803
http://hdl.handle.net/10138/306553
Thesis level: master's thesis
Degree program: Genetiikan ja molekulaaristen biotieteiden maisteriohjelma
Master's Programme in Genetics and Molecular Biosciences
Magisterprogrammet i genetik och molekylära biovetenskaper
Specialisation: Molekulaaristen ja analyyttisten biotieteiden opintosuunta
Molecular and Analytical Health Biosciences
Molekylärä och analytiska biovetenskaper
Abstract: Human umbilical vein endothelial cells are responsible for maintaining and forming new vessels from existing ones, in a biological process called sprouting angiogenesis. Sprouting angiogenesis is a crucial mechanism for the resolution of hypoxia and normal development of tissues. It also plays a key role in internal plague hemorrhages, which can lead to embolisms and other cardiovascular complications. Angiogenesis is also crucial for cancer development. Sprouting angiogenesis is initiated by hypoxic tissue excreted vascular endothelial growth factor gradient, which induces normal endothelial cells into either a proliferative stalk cell or a signal sensing tip cell phenotype. Both of these cell types depend on the rapid flow of lipids to their plasma membrane, either to form plasma membrane protrusions in tip cells or as new plasma membrane material in dividing stalk cells. This flow is envisioned to involve both vesicle-mediated and non-vesicular mechanisms. A major non-vesicular route of lipid transfer occurs at membrane contact sites via lipid transport proteins. Furthermore, lipids can be transported to the plasma membrane by the direct fusion of vesicles or endosomes with the plasma membrane This thesis set out to explore the role of two membrane contact site proteins, oxysterol-binding protein- related protein 2 and protrudin, in angiogenesis and lipid transfer. Their role was examined by RNA-sequencing transient knock-down samples of these proteins in HUVECs. The RNA-sequencing data was examined by differential expression, gene ontology overrepresentation and gene set enrichment analyses. Gene expression analysis provided almost 10 000 significantly changed transcripts (adjusted p-values < 0.05), in each silenced cell type. The distribution of differentially expressed genes in oxysterol-binding protein- related protein 2 silenced cells, is skewed toward negative fold changes, whereas the distribution of differentially expressed genes in protrudin silenced samples is normally distributed. The results also show significant changes in gene ontologies related to proliferation, cell cycle, angiogenesis as well as hypoxia in both sample types. Gene set enrichment analysis showed upregulation in angiogenesis related pathways, such as the PI3K-Akt and MAPK pathways, in both samples. Significant downregulation was present in cell cycle related pathways and cholesterol biosynthesis pathway in both ORP2 and protrudin silenced samples.
Subject: angiogeneesi
endoteelisolu
geenien ilmentyminen
ORP2
protrudiini
RNA-sekvensointi
kalvokontaktiproteiini
funktionaalinen genetiikka
RNA-sequencing
ORP2
protrudin
membrane contacts site
angiogenesis
endothelial cell
gene expression
functional genomics


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