Cytokine resistance in melanoma [Elektronische Ressource] / vorgelegt von Waraporn Komyod

Waraporn

Sujets

• Gesundheit

Informations

• Publié par: rheinisch-westfalischen_technischen_hochschule_-rwth-_aachen
• Publié le: 01 janvier 2007
• Nombre de lectures: 22
• Langue: Deutsch
• Poids de l'ouvrage: 2 Mo

Extrait

Cytokine resistance in melanoma



Von der Fakultät für Mathematik, Informatik und Naturwissenschaften
der Rheinisch-Westfälischen Technischen Hochschule Aachen
zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften
genehmigte Dissertation



vorgelegt von
Diplom-Biochemikerin
Waraporn Komyod
aus Phitsanulok, Thailand



Berichter: Universitätsprofessorin Dr. Iris Behrmann
Universitätsprofessor Dr. Fritz M. Kreuzaler


Tag der mündlichen Prüfung: 30. November 2007


Diese Dissertation ist auf den Internetseiten der Hochschulbibliothek online verfügbar.





































Publications from this work

1. Komyod, W., Böhm, M., Metze, D., Heinrich, P.C. & Behrmann, I. (2007)
Constitutive suppressor of cytokine signaling 3 expression confers a growth advantage
to a human melanoma cell line.
Mol Cancer Res 5, 271-281.

2. Komyod, W., Bauer, U.M., Heinrich, P.C., Haan, S. & Behrmann, I. (2005)
Are STATs arginine-methylated?
J Biol Chem 280, 21700-21705.

Further publications:

3. Behrmann, I., Wallner, S., Komyod, W., Heinrich, P.C., Schuierer, M., Buettner,
R. & Bosserhoff, A.K. (2003)
Characterization of methylthioadenosin phosphorylase (MTAP) expression in
malignant melanoma.
Am J Pathol 163, 683-690.

4. Behrmann, I., Smyczek, T., Heinrich, P.C., Schmitz-Van de Leur, H., Komyod,
W., Giese, B., Müller-Newen, G., Haan, S. & Haan, C. (2004)
Janus kinase (Jak) subcellular localization revisited: the exclusive membrane
localization of endogenous Janus kinase 1 by cytokine receptor interaction uncovers
the Jak/receptor complex to be equivalent to a receptor tyrosine kinase.
J Biol Chem 279, 35486-35493.

5. Kortylewski, M., Komyod, W., Kauffmann, M.E., Bosserhoff, A., Heinrich, P.C.
& Behrmann, I. (2004)
Interferon-gamma-mediated growth regulation of melanoma cells: involvement of
STAT1-dependent and STAT1-independent signals.
J Invest Dermatol 122, 414-422.
Table of content I
Table of content
Abbreviations........................................................................................................................... V
1 Introduction........................................................................................................................ 1
1.1 Progression of cutaneous melanoma...................................................................................... 1
1.2 IL-6-type cytokines .................................................................................................................... 4
1.2.1 Interleukin 6 (IL-6)......................................................................................................... 5
1.2.2 Oncostatin M (OSM)......................................................................................................5
1.3 Signaling pathways activated by IL-6-type cytokines........................................................ 6
1.3.1 Jak/STAT signaling pathway ......................................................................................... 6
1.3.2 Activation of MAPK cascades ....................................................................................... 8
1.4 Interferon signaling pathway .................................................................................................. 8
1.5 Negative regulation of the Jak/STAT signaling pathway ............................................... 10
1.5.1 Protein tyrosine phosphatases (PTPs) .......................................................................... 10
1.5.2 Protein inhibitor of activated STAT (PIAS) ................................................................ 11
1.5.3 Suppressors of cytokine signaling (SOCS) .................................................................. 11
1.6 Protein arginine methylation................................................................................................. 13
1.6.1 Protein arginine methyltransferases (PRMTs) ............................................................. 13
1.6.2 The role of protein arginine methylation in signal transduction .................................. 14
1.6.3 Methylation inhibitors..................................................................................................15
1.7 Cytokine responses in melanoma.......................................................................................... 16
1.8 Aims of this study ..................................................................................................................... 17
2 Materials and Methods .................................................................................................... 18
2.1 Materials..................................................................................................................................... 18
2.1.1 Chemicals and buffers 18
2.1.2 Radiochemicals............................................................................................................ 18
2.1.3 Prokaryotic cells........................................................................................................... 18
2.1.4 Eukaryotic cells............................................................................................................ 18
2.1.5 Media and reagents for cell culture.............................................................................. 19
2.1.6 Cytokines and soluble IL-6Rα ..................................................................................... 19
2.1.7 Inhibitors...................................................................................................................... 19
II Table of content
2.1.8 Enzymes....................................................................................................................... 19
2.1.9 Antibodies.................................................................................................................... 20
2.1.10 Plasmids....................................................................................................................... 21
2.1.11 Oligonucleotides........................................................................................................... 22
2.1.12 Commercial kits........................................................................................................... 23
2.2 Methods ...................................................................................................................................... 24
2.2.1 Culture of bacteria........................................................................................................ 24
2.2.2 Preparation of transformation-competent E. coli cells................................................. 24
2.2.3 Transformation of competent E. coli cells ................................................................... 24
2.2.4 Purification of plasmid DNA.......................................................................................25
2.2.5 Quantification of nucleic acids..................................................................................... 25
2.2.6 Restriction digestion of plasmid DNA......................................................................... 25
2.2.7 Agarose gel electrophoresis.........................................................................................25
2.2.8 DNA purification from agarose gels ............................................................................ 26
2.2.9 DNA ligation................................................................................................................ 26
2.2.10 Construction of expression vectors .............................................................................. 26
2.2.11 Automated DNA sequencing ....................................................................................... 27
2.2.12 Cell culture................................................................................................................... 28
2.2.13 Cell Transfection.......................................................................................................... 28
2.2.14 Generation of stable cell lines ...................................................................................... 29
2.2.15 Preparation of cell lysates ............................................................................................ 29
2.2.16 Determination of protein concentration according to Bradford................................... 29
2.2.17 Immunoprecipitations...................................................................................................30
2.2.18 SDS-polyacrylamide gel electrophoresis ..................................................................... 30
2.2.19 Western blotting........................................................................................................... 31
2.2.20 Immunodetection.......................................................................................................... 31
2.2.21 Flow cytometry