Interaction of human papillomavirus 20 and 4-nitroquinoline-1-oxide in carcinogenesis [Elektronische Ressource] / vorgelegt von Elsa Herráez Hernández

V4035ea

Sujets

• Gesundheit

Informations

• Publié par: ruprecht-karls-universitat_heidelberg
• Publié le: 01 janvier 2009
• Nombre de lectures: 20
• Poids de l'ouvrage: 55 Mo

Extrait

Aus dem Deutschen Krebsforschungszentrum Heidelberg
(Geschäftsführender Direktor: Prof. Dr. med. Otmar D. Wiestler)
Abteilung für Tumorvirus% Charakterisierung
(Abteilungsleiter: Prof. Dr. Ethel%Michele de Villeirs)





Interaction of human papillomavirus 20 and
4nitroquinoline1oxide in carcinogenesis



Inaugural dissertation
zur
Erlangung der Doktorwürde
Der
Naturwissenschaftlich%Mathematischen Gesamtfakultä t
der
Ruprecht % Karls % Universität
Heidelberg


vorgelegt von
Elsa Herráez Hernández

aus
Ávila, Spanien
2009


Dissertation
submitted to the
Combined Faculties for the Natural Sciences and for Mathematics
of the Ruperto%Carola University of Heidelberg, Gemr any
for the degree of
Doctor of Natural Sciences
















presented by
Elsa Herráez Hernández
born in Ávila, Spain
Oral%examination: ................................................
































Referees: (Herr) Prof. Dr. G.J. Hämmerling.
(Herr) Prof. Dr. P. Angel.





























To Lilín Table of contents

Table of contents
Abbreviations ......................................................................................................vi
Summary .............................................................................................................ix
Zusammenfassung ..............................................................................................xi
1 Introduction ....................................................................................................13
1.1 Papillomaviruses ........................................................................................13
1.1.1 General aspects .......................................................................................................13
1.1.2 HPV types ................................................................................................................14
1.1.2.1 Mucosal types...................................................................................................14
1.1.2.2 Cutaneous types................................................................................................14
1.1.3 HPV20 .....................................................................................................................15
1.1.4 HPV genomic organization .....................................................................................15
1.1.4.1 Early proteins....................................................................................................17
1.1.4.2 Late proteins .....................................................................................................22
1.1.4.3 Upstream regulatory region ..............................................................................23
1.1.5 HPV life cycle ..........................................................................................................23
1.1.6 HPV and cancer.......................................................................................................25
1.1.6.1 Cervical cancer .................................................................................................25
1.1.6.2 Head and neck cancer .......................................................................................26
1.1.6.3 Non melanoma skin cancer...............................................................................27
1.2 4%NQO......................................................................................................28
1.3 p53 gene family..........................................................................................29
1.3.1 p53 ...........................................................................................................................29
1.3.2 p63 ...........................................................................................................................30
1.4 Cell cycle control by cyclin%dependent kinase nihibitors..........................31
1.5 The skin......................................................................................................31
1.5.1 Skin organization.....................................................................................................31
1.5.2 Epidermal layers......................................................................................................32
1.5.3 The skin barrier .......................................................................................................33
1.5.4 Keratins ...................................................................................................................34
1.5.4.1 Keratin expression in simple epithelia..............................................................35
1.5.4.2 Keratin expression in stratified epithelia..........................................................35 Table of contents

1.5.4.3 Keratin expression in squamous cell carcinoma...............................................36
1.5.5 Differentiation markers ...........................................................................................37
1.5.5.1 Involucrin..........................................................................................................37
1.5.5.2 Loricrin .............................................................................................................37
1.6 Aim of the project ......................................................................................37
2 Materials..........................................................................................................39
2.1 Equipment ..................................................................................................39
2.2 Consumables ..............................................................................................41
2.3 Reagents .....................................................................................................42
2.4 Software .....................................................................................................46
2.5 Commercial kits .........................................................................................46
2.6 Solutions.....................................................................................................47
2.7 Bacteria.......................................................................................................52
2.7.1 Strain .......................................................................................................................52
2.7.2 Medium ....................................................................................................................53
2.8 Mammalian cell lines .................................................................................53
2.8.1 Cell lines and mediums............................................................................................53
2.9 Plasmids .....................................................................................................55
2.10 Primers .....................................................................................................56
2.11 Enzymes ...................................................................................................58
2.12 Antibodies ................................................................................................58
2.13 Markers.....................................................................................................59
3 Methods ...........................................................................................................60
3.1 Bacterial manipulation ...............................................................................60
3.1.1 Bacterial culture......................................................................................................60
3.1.2 Bacterial stocks........................................................................................................60
3.1.3 Bacterial transformation .........................................................................................60
3.2 DNA ...........................................................................................................61
3.2.1 Small scale preparation...........................................................................................61
3.2.2 Large scale preparation ..........................................................................................61
3.2.3 Determination of nucleic acid concentration ..........................................................62 Table of contents

3.2.4 Agarose gel electrophoresis ....................................................................................62
3.2.4.1 DNA isolation from agarose gels .....................................................................62
3