The role of 17beta-Estradiol on 2,3,7,8-Tetrachlorodibenzo-p-dioxin-mediated axidative DNA damage in liver cell models [Elektronische Ressource] / vorgelegt von Manuela Göttel

Benjamin Franklin

Informations

• Publié par: technische_universitat_kaiserslautern
• Publié le: 01 janvier 2009
• Nombre de lectures: 46
• Langue: Deutsch
• Poids de l'ouvrage: 2 Mo

Extrait

THE ROLE OF 17BETA-ESTRADIOL ON
2,3,7,8-TETRACHLORODIBENZO-P-DIOXIN-
MEDIATED OXIDATIVE DNA DAMAGE IN
LIVER CELL MODELS


dem Fachbereich Chemie der Universität Kaiserslautern
zur Erlangung des akademischen Grades
“Doktor der Naturwissenschaften”
eingereichte Dissertation

(D386)



vorgelegt von
Diplom Lebensmittelchemikerin
Manuela Göttel








Betreuung:

Prof. Dr. Dr. Dieter Schrenk,
Technische Universität Kaiserslautern


Der experimentelle Teil der vorliegenden Arbeit entstand im Zeitraum von Mai 2005 bis
Dezember 2008 im Fachbereich Chemie, Fachrichtung Lebensmittelchemie und Toxikolo-
gie an der Technischen Universität Kaiserslautern sowie in der Lebensmitteltoxikologie an
der Universität von Burgund in Dijon.















Eröffnung des Promotionsverfahrens: 07. Januar 2009
Tag der wissenschaftlichen Aussprache: 10. Dezember 2009

Prüfungskommision:
Vorsitzender: Prof. Dr. Werner Thiel
1. Berichterstatter: Prof. Dr. Dr. Dieter Schrenk
2. Berichterstatter: Prof. Dr. Marie-Christine Chagnon


ACKNOWLEDGEMENTS
First of all, I greatly want to thank my principal supervisor Prof. Dr. Dr. Dieter Schrenk for his support and the
opportunity to perform my doctoral thesis in his lab and work on such an interesting toxicological topic. The
presentation of my research results during international toxicology congresses such as EUROTOX or SOT were
a great experience. I also want to thank Prof. Schrenk for supporting my wish to organize a stay abroad to
perform parts of my research work in France.
Likewise, I profoundly want to thank Prof. Dr. Marie-Christine Chagnon at the laboratory of Food Toxicology
at the University of Burgundy in Dijon, France for having accepted me to work in her research group for
several months. The possibility to establish cooperation between the two labs even afterwards further added
to the immeasurable experience. I am thankful for her support, extensive advice, and fruitful discussions.
I would also like to thank Prof. Dr. Werner Thiel for accepting the responsibility of leading the board of ex-
aminers.
Thanks to all members of Prof. Schrenk’s, Prof. Eisenbrand’s, and Prof. Richling’s research groups for the nice
working atmosphere and the exchange and cooperation between the groups. I want to thank all the col-
leagues for the great time we spent together, even outside the lab. Especially, I want to thank my friends
Yvonne Fery, Yufanyi Ngiewih, Nicole Raquet, and Sven Triebel with whom I shared a lot of fun at the lab and
beyond.
Special thanks to our technicians for their expert technical support: Monika Gross for her record-worthy
isolation of rat hepatocytes and her organizing talent, Andreas Kamychnikow for his Best Western support,
and Wilja Daub for any kind of technical assistance.
Thanks to Anita Strohauer for her friendliness and help with any kind of problems. Thanks as well to Rainer
Scheuermann, who continuously spread his good humor in the lab, keep your smile! Thanks to Ari Vetter for
his availability when technical problems occured, especially with the comet software. Thanks to Sylvia
Schmidt, without your company in the comet lab many of the endless comet assay hours would have been
much less exciting, and to Phillip Bellion for the helpful discussions on the GSH assay.
Many thanks to all my internship students for their contribution to this work in any way: Christina Pachel,
Maria Tsaneva, Martin Chopra, and Estelle Renard.
Merci beaucoup to all the colleagues and friends in Dijon. Je vous remercie énormément for having welcomed
and integrated me in such a friendly fashion. It was always a great pleasure working with you.
In particular I would like to thank Isabelle Severin for her valuable and extensive advice on genotoxicity tests,
Coralie Dumont for her friendship and support in the transfection assays, Ludovic LeCorre for the fruitful
discussions, especially during the last months, and Laurence Dahbi for the numerous scientific discussions.
Catherine Berges and Willy Potier deserve my gratefulness, among other things, for helping me out with dry
ice for the transport of the uncounted samples that travelled between Kaiserslautern and Dijon. Many thanks
to all of you for the pauses café conviviales, which I always enjoyed.
I also want to express many thanks to Hélène Poirier at the laboratory of Nutrition at the University of Bur-
gundy in Dijon for providing me deep insights into the method of qPCR, for her time and patience, and many
helpful ideas.
My best wishes to Prof. Bernd Epe at the laboratory of pharmacy at the University of Mainz, who kindly
-/- mut/mut
provided us with repair-deficient OGG1 /csb mice. Special thanks to Markus Fussher, who performed
the alkaline elution technique with me, for his support and help.
Dr. Karl-Heinrich Fritzemeier (Bayer Schering, Germany) has to be thanked for kindly providing us with the
anti-estrogen ZK 191 703 and for answering my questions.
I would like to acknowledge the financial support in the award of a Doctoral Scholarship (Landesgraduier-
tenförderungsgesetz (LGFG) Rheinland-Pfalz) and a Foreign Exchange Scholarship (Deutscher Akademischer
Austauschdienst (DAAD)).
I am most grateful to my friends, especially to Anja and Jamie, for encouraging me throughout this work, for
their support and motivation.
Thanks to Portugal. The Man, Muse, and Black Rust for the inspiring music.
Without the constant support and help of my parents and my family this doctoral thesis would not have been
possible.
Special thanks to Benjamin: your support and encouragement always kept me going!
LIST OF ABBREVIATIONS

[3H]-thymidine tritium-labelled thymidine
µg microgram [unit]
µL microliter [unit]
µM Micromolar
17beta-HSD 17β-hydroxysteroid dehydrogenase
2-AAF 2-acetylaminofluorene
2-OHE2/4-OHE2 2-/4-hydroxyestradiol
8-MOP 8-methoxypsoralen
8-oxo-dG 8-hydroxy-2’-deoxyguanosine
3-MC 3-methylcholanthrene
Ade Adenine
AF activation functions
aFGF acidic fibroblast growth factor
AhR aryl hydrocarbon receptor
AhRC aryl hydrocarbon receptor complex
AhRR aryl hydrocarbon receptor repressor
AMP adenosine monophosphate
AR autoradiography
ARNT aryl hydrocarbon nuclear translocator
ATP adenosine triphosphate
ATSDR Agency for Toxic Substances and Disease Registry
BaP benzo(a)pyrene
BER base excision repair
beta-galactosidase β-gal
bHLH basic-helix-loop-helix
bp base pair
Brg1 Brahma/SWI2-related gene 1
BROD benzyloxyresorufin O-debenzylase
BSA bovine serum albumin
bw body weight
cAMP cyclic adenosine monophosphate
CBP cAMP response element binding protein
CE catechol estradiol
cDNA complementary DNA
ChIP chromatin immunoprecipitation
CHO chinese hamster ovary cells
COMT catechol-O-methyltransferase
CPRG chlorophenol-red β-D-galactopyranoside
csb cockayne syndrome B
CT treshold cycle
CYP cytochrome P450-dependent mono-oxygenases
DBD DNA binding domain
DCF 2’,7’-dichlorofluorescein
dCMP deoxycytidine monophosphate
DEN diethyl-N-nitrosamine
dG deoxyguanosine
DMEM Dulbecco’s modified Eagle’s medium
DMSO dimethylsulfoxide
DNA deoxyribonucleicacid
dNTP deoxynucleotide triphosphate
DRIP vitamin D receptor interacting protein
dRPase deoxyribophosphodiesterase
DSB double-strand break
dsDNA double-stranded DNA
DTNB 5,5’-dithiobis-2-nitrobenzoic acid
E.coli escherichia coli
e.g. for example
E1 estrone
E2 17beta- estradiol
E3 estriol
EGF epidermal growth factor
Endo endonuclease
EndRet endoplasmatic reticulum
ER estrogen receptor
ERE estrogen response elements
ERE sponse element
EROD 7-ethoxyresorufin-O-deethylase
et al. and others
EtOH ethanol
EtRes 7-ethoxyresorufin
FAD flavin adenine dinucleotide
Fapy formamidopyrimidine/4,6-diamino-5-formamidopyrimidine/2,6-
diamino-4-hydroxy-5-formamidopyrimidine
FBS fetal bovine serum
Fen1 flap endonuclease I
FMN flavin mononucleotide.
FPG Formamidopyrimidine(Fapy)-DNA glycosylase
GC-MS gas chromatography mass spectrometry
GPx glutathione peroxidases
GR gluthatione reductase
GSH glutathione (reduced form)
GSSG oxidized glutathione/glutathione disulfide
GST glutathione-S-transferases
Gua guanine
Gy gray [unit]
h hour
h .../r ... human .../rat ...
HDCFDA 2’, 7’-dichlorodihydrofluorescein diacetate 2
HAH halogenated aromatic hydrocarbon
HAP1 human 5’AP-endonuclease
HAT histone acetyltransferase
HGF hepatocytes growth factor
HhH helix-hairpin-helix
HPLC-ECD high performance liquid chromatography with electrochemical
detection
HSD hydroxysteroid dehydrogenase
HSP heat shock protein
i.e. that is
i.p. intraperitoneal
I3C indole-3-carbinol
IARC Internation