{"id":{"repo_id":"freiburg-diss","oai_identifier":"oai:freidok.uni-freiburg.de:2481"},"canonical_url":"https://search.dev.ndltd.org/etd/freiburg-diss/oai:freidok.uni-freiburg.de:2481","repository":{"repo_id":"freiburg-diss","name":"University of Freiburg","base_url":"https://freidok.uni-freiburg.de/oai/oai2.php"},"display":{"title":"Posttranscriptional gene silencing of p27 in primary prostate epithelial cells by small inhibitory RNA (siRNA)","abstract":"The cell cycle is an ordered set of events, culminating in cell growth and division into two daughter cells. The cell cycle is regulated by the intricate interplay of many molecules. Cdks (cyclin dependent kinases), along with cyclins, are major control switches for the cell cycle, causing the cell to move from G1 to S or G2 to M. Their function is tightly regulated by Cdk-inhibitors such as the p21 and p27 Cip/Kip proteins. Defects in many of the molecules that regulate the cell cycle have been implicated in cancer development. An important example is the common downregulation of the Cdk inhibitor p27 in prostate cancers. p27/Kip1 regulates the cell cycle by inhibiting the checkpoint kinase Cdk2/cyclin E kinase and blocking cell cycle progression in G1. In prostate cancer, reduced levels of p27 protein are associated with clinically aggressive tumors and poor prognosis. However, the molecular mechanisms of how the decrease in p27 levels contributes to prostate tumorigenesis are still poorly understood. <br>The aim of this study was to establish a system to downregulate p27 protein in primary diploid prostate epithelial cells (PrEC) for prolonged periods in order to create a model for further studies on its potential role in prostate cancer. In the experiments described here, sustained long term knockdown of p27 was achieved in human primary prostate cells by designing a lentivirus based vector system to deliver p27 specific siRNA. Immunoblotting showed a striking though not complete downregulation of p27 in PrEC. This may imitate p27 downregulation in tumorigenesis even better than a complete genetic knockout as the p27 gene is virtually never homozygously inactivated in prostate cancers. Thus, this system should provide a suitable model for future studies addressing the potential role of p27 in human prostate cancer development.","abstract_html":"The cell cycle is an ordered set of events, culminating in cell growth and division into two daughter cells. The cell cycle is regulated by the intricate interplay of many molecules. Cdks (cyclin dependent kinases), along with cyclins, are major control switches for the cell cycle, causing the cell to move from G1 to S or G2 to M. Their function is tightly regulated by Cdk-inhibitors such as the p21 and p27 Cip/Kip proteins. Defects in many of the molecules that regulate the cell cycle have been implicated in cancer development. An important example is the common downregulation of the Cdk inhibitor p27 in prostate cancers. p27/Kip1 regulates the cell cycle by inhibiting the checkpoint kinase Cdk2/cyclin E kinase and blocking cell cycle progression in G1. In prostate cancer, reduced levels of p27 protein are associated with clinically aggressive tumors and poor prognosis. However, the molecular mechanisms of how the decrease in p27 levels contributes to prostate tumorigenesis are still poorly understood. &lt;br&gt;The aim of this study was to establish a system to downregulate p27 protein in primary diploid prostate epithelial cells (PrEC) for prolonged periods in order to create a model for further studies on its potential role in prostate cancer. In the experiments described here, sustained long term knockdown of p27 was achieved in human primary prostate cells by designing a lentivirus based vector system to deliver p27 specific siRNA. Immunoblotting showed a striking though not complete downregulation of p27 in PrEC. This may imitate p27 downregulation in tumorigenesis even better than a complete genetic knockout as the p27 gene is virtually never homozygously inactivated in prostate cancers. Thus, this system should provide a suitable model for future studies addressing the potential role of p27 in human prostate cancer development.","abstract_has_math":false,"creators":["Mertelsmann-Voss, Christina"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Veelken, Hendrik"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":null,"date_issued":"","date_published":null,"updated_at":"2026-07-24T02:22:46Z","subjects":["siRNA","PrEC","p27","Zellzyklus","Prostate Cancer, Posttranscriptional Gene Silencing"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://freidok.uni-freiburg.de/data/2481","outbound_label":"Repository record","outbound_source":"source_url"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Veelken, Hendrik"]},{"key":"dc:creator","label":"Author","values":["Mertelsmann-Voss, Christina"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:type","label":"Dc Type","values":["DoctoralThesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["siRNA","PrEC","p27","Zellzyklus","Prostate Cancer, Posttranscriptional Gene Silencing"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["The cell cycle is an ordered set of events, culminating in cell growth and division into two daughter cells. The cell cycle is regulated by the intricate interplay of many molecules. Cdks (cyclin dependent kinases), along with cyclins, are major control switches for the cell cycle, causing the cell to move from G1 to S or G2 to M. Their function is tightly regulated by Cdk-inhibitors such as the p21 and p27 Cip/Kip proteins. Defects in many of the molecules that regulate the cell cycle have been implicated in cancer development. An important example is the common downregulation of the Cdk inhibitor p27 in prostate cancers. p27/Kip1 regulates the cell cycle by inhibiting the checkpoint kinase Cdk2/cyclin E kinase and blocking cell cycle progression in G1. In prostate cancer, reduced levels of p27 protein are associated with clinically aggressive tumors and poor prognosis. However, the molecular mechanisms of how the decrease in p27 levels contributes to prostate tumorigenesis are still poorly understood. <br>The aim of this study was to establish a system to downregulate p27 protein in primary diploid prostate epithelial cells (PrEC) for prolonged periods in order to create a model for further studies on its potential role in prostate cancer. In the experiments described here, sustained long term knockdown of p27 was achieved in human primary prostate cells by designing a lentivirus based vector system to deliver p27 specific siRNA. Immunoblotting showed a striking though not complete downregulation of p27 in PrEC. This may imitate p27 downregulation in tumorigenesis even better than a complete genetic knockout as the p27 gene is virtually never homozygously inactivated in prostate cancers. Thus, this system should provide a suitable model for future studies addressing the potential role of p27 in human prostate cancer development.","Der Zellzyklus ist eine streng koordinierte Folge von Ereignissen, in deren Folge es zu Wachstum und schlussendlich zur Teilung der jeweiligen Zelle in zwei Tochterzellen kommt. Die Kontrolle der genauen Abfolge des Zellzyklus wird durch das komplexe Zusammenspiel einer Vielzahl regulatorischer Moleküle ermöglicht. Cdk (cyclin dependent kinases), zusammen mit Cyklinen, tragen übergeordnete Kontrollfunktionen und bewirken so das Fortschreiten der Zelle innerhalb des Zellzyklus von der G1 zur S Phase oder auch von G2 zu M. Die Aktivität der Cdk wird streng reguliert von sogenannten Cdk-Inhibitoren wie dem p21- und p27-Cip/Kip Protein. Defekte innerhalb solcher Regulationsproteine werden heute als wichtiger Schritt in der Krebsentstehung gewertet. Ein Beispiel hierfür ist die Herunterregulierung der Expression des Cdk Inhibitors p27 im Rahmen der Entstehung von Prostata Karzinomen. p27/Kip1 reguliert den Zell Zyklus, indem es die Checkpoint Kinase Cdk2/cyclin E inhibiert und somit die Progression der Zelle in die G1 Phase hemmt. Beim Prostata Karzinom sind erniedrigte p27 Protein Level mit klinisch aggressiven Tumoren und schlechter Prognose assoziiert. Der genaue Mechanismus hinter diesem Phänomen ist jedoch bisher nur schlecht verstanden. <br>Das Ziel dieser Arbeit war es, ein System zu entwickeln, in dem p27 Protein Level in primär diploiden Prostata Epithelzellen (primary diploid prostate epithelial cells , PrEC) künstlich über lange Zeit supprimiert werden können, um ein Modell für das Studium der Entstehung des Prostata Carcinoms zu schaffen. In den hier beschriebenen Experimenten wurde eine dauerhafte Suppression von p27 Leveln mit Hilfe eines lentiviral basierten Vektor Systems zur Übertragung von p27 spezifischer siRNA erreicht. Immunoblots zeigten eine deutliche, wenn auch nicht komplette Herunterregulierung von p27 in PrEC. Dies könnte die p27 Suppression bei der Entstehung von Prostatakrebs sogar besser imitieren als ein kompletter genetischer Knockout, da bei diesen Tumoren das p27 Gen praktisch niemals homozygot inaktiviert wird. Das von uns entwickelte System sollte also ein adäquates Modell zur Erforschung der Rolle von p27 in der Entstehung humaner Prostata Karzinome liefern."]},{"key":"dc:format.medium","label":"Dc Format Medium","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Posttranscriptional gene silencing of p27 in primary prostate epithelial cells by small inhibitory RNA (siRNA)","Posttranskriptionelles Gene Silencing von p27 in primären Prostata Epithelzellen mit Hilfe von small inhibitory RNA (siRNA)"]}]}],"canonical_facts":{"dc:contributor":["Veelken, Hendrik"],"dc:creator":["Mertelsmann-Voss, Christina"],"dc:description.abstract":["The cell cycle is an ordered set of events, culminating in cell growth and division into two daughter cells. The cell cycle is regulated by the intricate interplay of many molecules. Cdks (cyclin dependent kinases), along with cyclins, are major control switches for the cell cycle, causing the cell to move from G1 to S or G2 to M. Their function is tightly regulated by Cdk-inhibitors such as the p21 and p27 Cip/Kip proteins. Defects in many of the molecules that regulate the cell cycle have been implicated in cancer development. An important example is the common downregulation of the Cdk inhibitor p27 in prostate cancers. p27/Kip1 regulates the cell cycle by inhibiting the checkpoint kinase Cdk2/cyclin E kinase and blocking cell cycle progression in G1. In prostate cancer, reduced levels of p27 protein are associated with clinically aggressive tumors and poor prognosis. However, the molecular mechanisms of how the decrease in p27 levels contributes to prostate tumorigenesis are still poorly understood. <br>The aim of this study was to establish a system to downregulate p27 protein in primary diploid prostate epithelial cells (PrEC) for prolonged periods in order to create a model for further studies on its potential role in prostate cancer. In the experiments described here, sustained long term knockdown of p27 was achieved in human primary prostate cells by designing a lentivirus based vector system to deliver p27 specific siRNA. Immunoblotting showed a striking though not complete downregulation of p27 in PrEC. This may imitate p27 downregulation in tumorigenesis even better than a complete genetic knockout as the p27 gene is virtually never homozygously inactivated in prostate cancers. Thus, this system should provide a suitable model for future studies addressing the potential role of p27 in human prostate cancer development.","Der Zellzyklus ist eine streng koordinierte Folge von Ereignissen, in deren Folge es zu Wachstum und schlussendlich zur Teilung der jeweiligen Zelle in zwei Tochterzellen kommt. Die Kontrolle der genauen Abfolge des Zellzyklus wird durch das komplexe Zusammenspiel einer Vielzahl regulatorischer Moleküle ermöglicht. Cdk (cyclin dependent kinases), zusammen mit Cyklinen, tragen übergeordnete Kontrollfunktionen und bewirken so das Fortschreiten der Zelle innerhalb des Zellzyklus von der G1 zur S Phase oder auch von G2 zu M. Die Aktivität der Cdk wird streng reguliert von sogenannten Cdk-Inhibitoren wie dem p21- und p27-Cip/Kip Protein. Defekte innerhalb solcher Regulationsproteine werden heute als wichtiger Schritt in der Krebsentstehung gewertet. Ein Beispiel hierfür ist die Herunterregulierung der Expression des Cdk Inhibitors p27 im Rahmen der Entstehung von Prostata Karzinomen. p27/Kip1 reguliert den Zell Zyklus, indem es die Checkpoint Kinase Cdk2/cyclin E inhibiert und somit die Progression der Zelle in die G1 Phase hemmt. Beim Prostata Karzinom sind erniedrigte p27 Protein Level mit klinisch aggressiven Tumoren und schlechter Prognose assoziiert. Der genaue Mechanismus hinter diesem Phänomen ist jedoch bisher nur schlecht verstanden. <br>Das Ziel dieser Arbeit war es, ein System zu entwickeln, in dem p27 Protein Level in primär diploiden Prostata Epithelzellen (primary diploid prostate epithelial cells , PrEC) künstlich über lange Zeit supprimiert werden können, um ein Modell für das Studium der Entstehung des Prostata Carcinoms zu schaffen. In den hier beschriebenen Experimenten wurde eine dauerhafte Suppression von p27 Leveln mit Hilfe eines lentiviral basierten Vektor Systems zur Übertragung von p27 spezifischer siRNA erreicht. Immunoblots zeigten eine deutliche, wenn auch nicht komplette Herunterregulierung von p27 in PrEC. Dies könnte die p27 Suppression bei der Entstehung von Prostatakrebs sogar besser imitieren als ein kompletter genetischer Knockout, da bei diesen Tumoren das p27 Gen praktisch niemals homozygot inaktiviert wird. Das von uns entwickelte System sollte also ein adäquates Modell zur Erforschung der Rolle von p27 in der Entstehung humaner Prostata Karzinome liefern."],"dc:format.medium":["application/pdf"],"dc:subject":["siRNA","PrEC","p27","Zellzyklus","Prostate Cancer, Posttranscriptional Gene Silencing"],"dc:title":["Posttranscriptional gene silencing of p27 in primary prostate epithelial cells by small inhibitory RNA (siRNA)","Posttranskriptionelles Gene Silencing von p27 in primären Prostata Epithelzellen mit Hilfe von small inhibitory RNA (siRNA)"],"dc:type":["DoctoralThesis"]},"updated_at":"2026-07-24T02:22:46Z"}