Dokument: Epigenetische Analyse von Biomarkerkandidaten auf ihre Methylierungsmuster im Prostatakarzinom
| Titel: | Epigenetische Analyse von Biomarkerkandidaten auf ihre Methylierungsmuster im Prostatakarzinom | |||||||
| Weiterer Titel: | Epigenetic analysis of biomarker candidates for their methylation patterns in prostate cancer | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=66679 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20240909-112941-0 | |||||||
| Kollektion: | Dissertationen | |||||||
| Sprache: | Deutsch | |||||||
| Dokumententyp: | Wissenschaftliche Abschlussarbeiten » Dissertation | |||||||
| Medientyp: | Text | |||||||
| Autor: | Sheikh, Jamal [Autor] | |||||||
| Dateien: |
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| Beitragende: | Prof. Dr. rer. nat. Santourlidis, Simeon [Gutachter] Prof. Dr. med. Krieg, Andreas [Gutachter] | |||||||
| Stichwörter: | Epigenetik, Biomarker, Kandidatengene, Prostatakarzinom, DNA-Methylierung | |||||||
| Dewey Dezimal-Klassifikation: | 600 Technik, Medizin, angewandte Wissenschaften » 610 Medizin und Gesundheit | |||||||
| Beschreibungen: | Das Prostatakarzinom stellt in den entwickelten Ländern den häufigsten und weltweit den zweithäufigsten Tumor des Mannes dar. Die Diagnostik umfasst u.a. die Bestimmung des PSA-Werts. Dieser ist jedoch vermindert sensitiv und wenig spezifisch für das Prostatakarzinom. Aus den genannten Tatsachen erwächst eine besondere Wichtigkeit für die gezielte Diagnostik, um eine Überdiagnostik und -therapie abzuwenden. Ein vielversprechender Ansatz stellt hierbei das Feld der Epigenetik dar. Epigenetik meint dabei alle Mechanismen, die die Genexpression beeinflussen, ohne dabei die Basenabfolge der DNA selbst zu verändern. Unter anderem ist die Methylierung von CpG-Inseln in Promotorregionen ein Mechanismus, welcher diagnostisches Potenzial birgt. Es wurde bereits in methylierungsspezifischen Arrays beobachtet, dass bestimmte CpG-Inseln in pathologisch begutachteten gesunden Gewebeproben der Prostata eher hypermethyliert sind, während sie beim Prostatakarzinom hypomethyliert sind. Für die vorliegende Arbeit wurde je ein Genlocus des XIST-Gens (x inactivation specific transcript) und des KIR2DL3-Gens (transmembranäres Protein auf der Oberfläche von Natürlichen Killerzellen) auf ihren Methylierungsstatus in mehreren frei verkäuflichen, prostataassoziierten Zelllinien sowie in der DNA, isoliert aus insgesamt sechs Primärproben aus Prostatatumorgeweben, analysiert. Ziel war es, den Methylierungsstatus von Prostatatumorzellen und pathologischen nicht-malignen Zellen der Prostata graphisch abzubilden und zu quantifizieren.
Die wesentlichen Schritte umfassten dabei zunächst die Bisulfitkonvertierung der aus Zellkulturen isolierten Zelllinien-DNA. Die relevanten Abschnitte der genannten Genloci wurden in vitro amplifiziert und mittels TA-Klonierung in chemisch kompetenten E. coli eingeführt und vervielfältigt. Nach weiterer Vervielfältigung in Flüssigmedien wurden die Genloci sequenziert, um ein detailliertes Methylierungsprofil dieser Regionen zu erstellen. Ergebnis war, dass im KIR2DL3-Gen ein vorhandener Unterschied zwischen Tumor-DNA und Nicht-Tumor-DNA in sechs von elf Proben festgestellt werden konnte, während im XIST-Genlocus lediglich ein subtiler Unterschied nachweisbar war. Die genauen DNA-Methylierungsprofile dieser Regionen wurden erstmalig für das Prostatakarzinom erhoben. Diese Studie könnte einen ersten Schritt darstellen, um der Entwicklung eines potenten Biomarkers näherzukommen, jedoch sind hierfür weitere langfristig angelegte Studien notwendig. Ein vorhandener Unterschied in einer einzelnen CpG-Position könnte sich diagnostisch zunutze gemacht werden und wäre mittels Idiolokaler Normierung durch eine Methylierungsspezifische PCR (MSPCR) verlässlich und schnell nachweisbar.Prostate cancer is the most common male tumour in developed countries and the second most common tumour worldwide. Diagnostics include the determination of PSA levels. However, this is reduced sensitive and not very specific for prostate cancer. These facts make targeted diagnostics particularly important in order to avoid overdiagnosis and overtreatment. The field of epigenetics represents a promising approach in this matter. Epigenetics refers to all mechanisms that influence gene expression without changing the base sequence of the DNA itself. Among other things, the methylation of CpG islands in promoter regions is a mechanism with diagnostic potential. It has already been observed in methylation-specific arrays that certain CpG islands tend to be hypermethylated in pathologically reviewed non.-malignant prostate tissue samples, whereas they are rather hypomethylated in prostate carcinoma. For the present study, one gene locus each of the XIST gene (x inactivation specific transcript, important for X inactivation) and the KIR2DL3 gene (transmembrane protein found on the surface of natural killer cells) were analyzed for their methylation status in several freely available prostate-associated cell lines and DNA isolated from a total of six primary samples of prostate tumor tissue. The aim was to graphically depict and quantify the methylation status of prostate tumour cells and non-malignant cells of the prostate. The individual steps involved the bisulphite conversion of cell line DNA isolated from cell cultures. The relevant segments of these gene loci were amplified in vitro and amplified by TA cloning in chemically competent E. coli. After further amplification in liquid media, the gene loci were sequenced to generate a detailed methylation profile of these regions. The result was that a difference between tumour DNA and non-tumour DNA could be detected in six out of eleven DNA samples in the gene KIR2DL3, while a subtle difference was detectable in the XIST gene locus. The exact DNA methylation profiles of these regions were collected for the first time for prostate cancer. This study could be a first step towards the development of a potent biomarker, but further long-term studies are needed. An existing difference in a single CpG position could be used diagnostically and could be easily and quickly determined by means of idiolocal normalization of Methylation Specific PCR. | |||||||
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| Lizenz: |  Dieses Werk ist lizenziert unter einer Creative Commons Namensnennung 4.0 International Lizenz | |||||||
| Bezug: | 2 Jahre | |||||||
| Fachbereich / Einrichtung: | Medizinische Fakultät | |||||||
| Dokument erstellt am: | 09.09.2024 | |||||||
| Dateien geändert am: | 09.09.2024 | |||||||
| Promotionsantrag am: | 19.03.2024 | |||||||
| Datum der Promotion: | 29.08.2024 |
