Cancer proteomics of mouse serum and liver tissue samples to discover candidate biomarkers for hepatocellular carcinoma (HCC) in c-Myc transgenic mice [Elektronische Ressource] / Maria Stella Ritorto

Maria Stella Ritorto

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• Gesundheit

Informations

• Publié par: gottfried_wilhelm_leibniz_universitat_hannover
• Publié le: 01 janvier 2011
• Nombre de lectures: 61
• Langue: Deutsch
• Poids de l'ouvrage: 4 Mo

Extrait

Cancer proteomics of mouse serum and liver tissue samples
to discover candidate biomarkers for
hepatocellular carcinoma (HCC) in c-Myc transgenic mice

Von der Naturwissenschaftlichen Fakultät
der Gottfried Wilhelm Leibniz Universität Hannover
zur Erlangung des Grades
Doktorin der Naturwissenschaften

Dr. rer. nat.

genehmigte Dissertation von


Dottore in Farmacia
Maria Stella Ritorto
geboren am 27.09.1979 in Rom (Italien)








2011

Cancer proteomics of mouse serum and liver tissue samples to discover candidate biomarkers for HCC in c-Myc transgenic mice




















Referent: Prof. Dr. Juergen Borlak
Lehrstuhl fuer Pharmaco- und Toxikogenomik
Medizinische Hochschule Hannover
Abteilung Molekulare Medizin und Medizinische Biotechnologie
Fraunhofer Institut fuer Toxikologie und Experimentelle Medizin


Korreferent: Prof. Dr. Rer. Nat. Juergen Alves
Biophysikalische Chemie
Medizinische Hochschule Hannover


Tag der Promotion: 10. Februar 2011


iii
Die vorliegende Arbeit wurde in der Zeit von 2006 bis 2010 am Fraunhofer Institut für
Toxicologie und Experimentelle Medizin in Hannover in der Abteilung für Molekulare
und Medizinische Biotechnologie angefertigt.



















Erklärung:
Hiermit erkläre ich, dass ich diese Dissertation selbstständig verfasst und alle
benutzten Hilfsmittel und Quelle sowie gegebenenfalls die zu Hilfeleistung
herangezogene Institutionen vollständig angegeben habe.

Diese Dissertation wurde nicht schon als Masterarbeit, Diplomarbeit oder andere
Prüfungsarbeit verwendet.




Hannover, im November 2010







Acknowledgments













Cancer proteomics of mouse serum and liver tissue samples to discover candidate biomarkers for HCC in c-Myc transgenic mice


he work behind this dissertation has been an incredible journey and a monumental milestone in
my academic life. I could not have embarked on this expedition and travelled this far without the T
passionate and continued support of advisors, colleagues, friends and family.

First of all,
…………...I would like to express my gratitude to Professor Jürgen Borlak, my main dissertation
advisor and supervisor. Over the past four years, we had numerous conversations and electronic
exchanges that constantly improved and refined this work.
A big gratitude
……………goes to Professors Uwe Heinrich and Clemens Dasenbrock from Fraunhofer Institute for
Toxicology and Experimental Medicine who were of great importance in finalizing this work and make
it true.
Moreover, I thank
…………..Professors Jürgen Alves and Andreas Pich for their particular kindness in accepting to be
my Coreferent and third Examiner, as well as Prof Christoph Peterhaensel for moderating the defence.
…………..Professor Arndt Vogel and his collaborators Jutta Lamle and Silke Marhenke, for their
professional help in clinical studies.
………….Among my colleagues, Kathrin Hoffmeyer for genomic data, as well as Gudrun Muehlbauer
and Kathrin Eidenschink for the optimum technical support in IHC analysis.
In particular,
……………..I would like to thank Dott Ignazio Garaguso, Dr. Bijon Chatterji, Dr. Roman Halter as well
as Eckhard Belau which were more than colleagues and they gave colour to my hard task.

This extraordinary voyage would certainly have been less extraordinary and colorful if not for the
engaging and heartwarming time spent with my fantastic friends, Tatjana, Christine, Dietmar and
Luan, along to all my Italian and other European crew of friends: to them, I want to say thank for
supporting my dark moments and giving a spread of smiles. I will be thankful to them forever.

Vorrei ringraziare in particolare Ignazio, con cui ho condiviso tutto il dottorato e a cui devo molto,
poiché è grazie al suo supporto professionale che sono riuscita a raggiungere la meta.

Il ringraziamento piú grande é rivolto alla mia famiglia ed in particolare ai miei fantastici genitori, con
cui ho condiviso gioie e dolori di questo dottorato e senza cui non ce l´avrei mai potuta fare.








iii Cancer proteomics of mouse serum and liver tissue samples to discover candidate biomarkers for HCC in c-Myc transgenic mice





















To my parents, for their love
(Ai miei genitori, per il loro amore)


























iv















Contents
















Cancer proteomics of mouse serum and liver tissue samples to discover candidate biomarkers for HCC in c-Myc transgenic mice


ABSTRACT..............................................................................................................................1
KURZZUSAMMENFASSUNG .................................................................................................3
INTRODUCTION......................................................................................................................5
THEORY ..................................................................................................................................7
THE ROLE OF MYC TRANSCRIPTION FACTOR IN NORMAL BIOLOGY AND CANCER .................................................8
Myc biological functions................................................................................................................ 11
Myc and cancer............................................................................................................................. 12
HEPATOCELLULAR CARCINOMA16
Diagnosis of HCC ......................................................................................................................... 17
Treatment of HCC 18
The role of c-Myc in HCC....... 19
Biomarkers for HCC...................................................................................................................... 19
Animal models of HCC.................................................................................................................. 20
A c-Myc transgenic mouse model of HCC.................................................................................... 22
PROTEOMICS-BASED BIOMARKER DISCOVERY....................................................................................................23
AIMS OF THE DOCTORAL THESIS.....................................................................................29
APPLIED METHODS31
MANTAINANCE OF TRANSGENIC MICE................................................................................................................32
PATIENT CHARACTERISTICS ...............................................................................................................................32
TISSUE AND SERUM PROTEOME ANALYSIS..........................................................................................................34
Protein separation and quantitation by 2-DE................................................................................ 35
MALDI-TOF/TOF mass spectrometry........................................................................................... 39
VALIDATION OF PROTEIN REGULATIONS............................................................................................................41
Western immunoblotting ............................................................................................................... 41
Immunohistochemistry.................................................................................................................. 41
GENE EXPRESSION DATA OF REGULATED PROTEINS42
BIO