Macrophage-related interaction pathways in tumor stroma:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2012
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| Schlagworte: | |
| Online-Zugang: | kostenfrei https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120712-1094701-1-2 Inhaltsverzeichnis |
| Beschreibung: | Zsfassung in dt. Sprache |
| Beschreibung: | 100 S. Ill., graph. Darst. |
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Datensatz im Suchindex
| _version_ | 1804149401470369792 |
|---|---|
| adam_text | IMAGE 1
1 INTRODUCTION 1
1.1 COMPOSITION OF THE TUMOR STROMA 1
1.1.1 TUMOR STROMA ECM 2
1.1.2 CELLS O F T H E T U M O R STROMA 2
1.1.2.1 FIBROBLASTS 3
1.1.2.2 BLOOD VESSEL CELLS 3
1.1.2.3 CELLS O F T H E IMMUNE SYSTEM 4
1.2 TUMOR-ASSOCIATED MACROPHAGES 5
1.3 INFLAMMATION AND TUMORIGENESIS 7
1.4 TUMOR STROMA INTERACTION PATHWAYS 8
1.4.1 EGFR FAMILY SIGNALING 9
1.4.2 HGF/MET SIGNALING 12
1.4.3 STAT3 SIGNALING 14
1.5 TARGETING TUMOR-ASSOCIATED MACROPHAGES FOR CANCER THERAPY 16
2 SPECIFIC A I M S 1 8
3 MATERIAL A N D METHODS 1 9
3.1 MATERIAL SOURCES 19
3.1.1 LABORATORY HARDWARE 19
3.1.2 LABORATORY CHEMICALS A N D BIOCHEMICAL 2 0
3.1.3 CHEMICALS A N D O T H E R MATERIAL FOR SILAC A N D MS ANALYSIS 2 1
3.1.4 ENZYMES 2 1
3.1.5 KITS A N D O T H E R MATERIALS 2 2
3.1.6 GROWTH FACTORS A N D LIGANDS 22
3.1.7 MEDIA 2 3
3.1.7.1 BACTERIAL MEDIA 2 3
3.1.7.2 CELL CULTURE MEDIA 2 3
3.1.8 STOCK SOLUTIONS A N D COMMONLY USED BUFFERS 2 3
3.1.9 CELLS 2 6
3.1.9.1 HUMAN CELL LINES 26
HTTP://D-NB.INFO/1033910198
IMAGE 2
3.1.9.2 MOUSE CELL LINES I 2 6
3.1.9.3 PRIMARY H U M A N CELLS 27
3.1.9.4 PRIMARY MURINE MONOCYTES A N D MACROPHAGES 27
3.1.9.5 BACTERIAL STRAINS 27
3.1.10 ANTIBODIES 2 8
3.1.10.1 PRIMARY ANTIBODIES FOR WESTERN BLOT 2 8
3.1.10.2 PRIMARY ANTIBODIES FOR IMMUNOPRECIPITATION 29
3.1.10.3 SECONDARY ANTIBODIES FOR WESTERN BLOT 2 9
3.1.10.4 ANTIBODIES FOR ELISA 2 9
3.1.10.5 ANTIBODIES FOR FACS 3 0
3.1.10.6 BLOCKING ANTIBODIES 3 1
3.1.11 OLIGONUCLEOTIDES A N D PLASMIDS 3 1
3.1.11.1 PRIMARY VECTORS 3 1
3.1.11.2 CONSTRUCTS 32
3.1.11.3 SIRNA OLIGONUCLEOTIDES (HUMAN SPECIFIC) 32
3.1.11.4 PRIMERS FOR REAL-TIME PCR (HUMAN SPECIFIC) 3 2
3.1.11.5 PRIMERS FOR REAL-TIME PCR (MOUSE SPECIFIC) 3 3
3.1.12 SOFTWARE 3 4
3.2 METHODS 34
3.2.1 METHODS IN MOLECULAR BIOLOGY 3 4
3.2.1.1 CULTIVATION A N D MAINTENANCE O F BACTERIAL STRAINS 3 4
3.2.1.2 PREPARATION O F C O M P E T E N T BACTERIA 3 4
3.2.1.3 TRANSFORMATION O F C O M P E T E N T BACTERIA 3 5
3.2.1.4 AMPLIFICATION O F PLASMIDS 3 5
3.2.1.5 PLASMID PURIFICATION 3 5
3.2.1.6 RNA ISOLATION A N D CDNA SYNTHESIS 3 5
3.2.1.7 QUANTITATIVE REAL-TIME PCR ANALYSIS 3 6
3.2.2 M E T H O D S IN MAMMALIAN CELL CULTURE 3 6
3.2.2.1 GENERAL CELL CULTURE TECHNIQUES 3 6
3.2.2.2 GROWTH FACTOR STIMULATION A N D INHIBITOR T R E A T M E N T 3 6
3.2.2.3 ISOLATION O F PRIMARY HUMAN PERIPHERAL BLOOD MONOCYTES 3 7
3.2.2.4 ISOLATION O F PRIMARY MURINE PERITONEAL MACROPHAGES 3 7
3.2.2.5 ISOLATION O F PRIMARY MURINE SPLEEN MONOCYTES 3 8
3.2.2.6 ISOLATION O F PRIMARY MURINE B O N E MARROW MONOCYTES 3 8
3.2.2.7 T R E A T M E N T O F H U M A N MACROPHAGES WITH CANCER CELL
CONDITIONED MEDIUM 39
IMAGE 3
3.2.2.8 TREATMENT O F MURINE MONOCYTES/MACROPHAGES WITH CANCER CELL
CONDITIONED MEDIUM 3 9
3.2.2.9 COCULTURE EXPERIMENT 39
3.2.2.10 PREPARATION O F CELL CULTURE CONDITIONED MEDIA (CM) 4 0
3.2.2.11 TREATMENT O F CM WITH BLOCKING ANTIBODIES 4 0
3.2.2.12 PREPARATION O F H U M A N SERUM 4 0
3.2.2.13 HEAT INACTIVATION OF HUMAN S E R U M 4 1
3.2.2.14 PREPARATION OF GELATIN COATED PLATES 4 1
3.2.2.15 TRANSFECTION OF MAMMALIAN CELLS 4 1
3.2.3 PROTEIN ANALYTICAL M E T H O D S 4 2
3.2.3.1 LYSIS O F CELLS WITH TRITON X-100 LYSIS BUFFER 4 2
3.2.3.2 DETERMINATION O F PROTEIN CONCENTRATION IN CELL LYSATES 4 2
3.2.3.3 IMMUNOPRECIPITATION OF PROTEINS 4 2
3.2.3.4 SDS-POLYACRYLAMIDE-GELELECTROPHORESIS (SDS-PAGE) 4 3
3.2.3.5 WESTERN BLOTTING 4 3
3.2.3.6 IMMUNOBLOT DETECTION 4 3
3.2.3.7 STRIPPING 4 3
3.2.3.8 ELISA 4 4
3.2.4 ANALYSIS O F WHOLE CELL-BASED ASSAYS 4 4
3.2.4.1 MIGRATION ASSAY 4 4
3.2.4.2 MTT ASSAY 4 5
3.2.4.3 FACS ANALYSES 4 5
3.2.5 SILAC EXPERIMENTS A N D MS ANALYSIS 4 6
3.2.5.1 CELL CULTURE IN SILAC MEDIUM 4 6
3.2.5.2 STABLE ISOTOPE LABELING, CELL LYSIS A N D ANTI-EGFR
IMMUNOPRECIPITATION FOR MASS SPECTROMETRY 4 6
3.2.5.3 MASS SPECTROMETRY ANALYSIS 4 6
RESULTS 4 7
1 HGF-MEDIATED EGFR PARACRINE CROSSTALK 47
4.1.1 HGF-INDUCED EGFR LIGAND RELEASE 4 7
4.1.1.1 IDENTIFICATION OF HGF-INDUCED EGFR LIGANDS IN SCC9 CELLS 4 7
4.1.1.2 HGF-INDUCED EGFR LIGAND RELEASE IS A COMMON CHARACTERISTIC O F
CANCER CELLS A N D D E P E N D S O N N E W
PROTEIN SYNTHESIS 4 9
4.1.1.3 METALLOPROTEASE-DEPENDENT SHEDDING O F EGFR LIGANDS 5 1
4.1.1.4 CM OF A MONOCYTIC CELL LINE INDUCES EGFR LIGAND INDUCTION VIA
HGF/MET 52
IMAGE 4
4.1.1.5 IDENTIFICATION O F T H E PATHWAY UNDERLYING EGF LIGAND INDUCTION
52
4.1.1.6 TGFO IS A STRONG INDUCER O F DIRECTED CELL MIGRATION IN SCC9
CELLS 5 4
4.1.2 HGF-INDUCED EGFR NEGATIVE REGULATION 5 5
4.1.2.1 HGF IS A PARACRINE INHIBITOR O F CLASSICAL EGFR ACTIVATION 5 5
4.1.2.2 LOSS OF EGFR TK ACTIVITY IS I N D E P E N D E N T OF A N
AUTOCRINE EGFR FEEDBACK LOOP 5 5
4.1.2.3 PATHWAY UNDERLYING EGFR TK BLOCKAGE 5 7
4.1.2.4 LOSS O F EGFR TK ACTIVITY IS INDEPENDENT O F T H E EGFR NEGATIVE
REGULATOR MIG6 5 7
4.1.2.5 HGF LEADS T O IMPAIRED EGFR-MEDIATED MIGRATION 5 8
4.1.2.6 HGF INDUCES VARIOUS EGFR INTERACTIONS 5 9
4.1.2.7 HGF LEADS T O E N H A N C E D SURVIVAL UPON GEFITINIB T R E A T
M E N T 62
4.1.2.8 HGF-INDUCED EGFR INTERACTIONS A R E RESISTANT T O GEFITINIB T R
E A T M E N T 6 3
4.1.2.9 M E T TK-INHIBITION RESTORES GEFITINIB SENSITIVITY 6 4
4.1.2.10 HGF-INDUCED EGFRTK BLOCKAGE IS A CHARACTERISTIC O F CANCER
CELLS 6 5
4.1.2.11 SIMILAR T O EGFR, ALSO HER2 A N D HER3 ACTIVATION IS BLOCKED
UPON HGF T R E A T M E N T 6 6
4.2 INTERACTION PATHWAYS BETWEEN BREAST CANCER CELLS AND MACROPHAGES IN
HUMAN AND MOUSE 67
4.2.1 INTERACTION PATHWAYS B E T W E E N BREAST CANCER CELLS A N D H U M
A N MACROPHAGES 6 7
4.2.1.1 CHARACTERIZATION OF H U M A N MACROPHAGES 6 7
4.2.1.2 HB-EGF A N D OSM RELEASE IN TUMOR-EDUCATED MACROPHAGES 6 7
4.2.1.3 MORPHOLOGY CHANGE O F H U M A N MACROPHAGES AFTER T R E A T M E
N T WITH CANCER CELL CM 7 0
4.2.2 GENERATING A N IN VITRO M O U S E MODEL 7 0
4.2.2.1 SCREEN FOR M O U S E REPORTER CELL LINES 7 0
4.2.2.2 CHARACTERIZATION O F M O U S E PERITONEAL MACROPHAGES 72
4.2.2.3 MORPHOLOGY CHANGE O F M O U S E MACROPHAGES AFTER T R E A T M E
N T WITH CANCER CELL CM 7 2
4.2.2.4 OSM RELEASE IN TUMOR-EDUCATED M O U S E MACROPHAGES 7 2
4.2.2.5 OSM TRANSCRIPT A N D EGFR LIGAND TRANSCRIPT INDUCTION IN
TUMOR-EDUCATED MACROPHAGES 74
4.2.2.6 OSM RELEASE IN B O N E MARROW A N D SPLEEN-DERIVED MONOCYTES 7 6
5 DISCUSSION 7 8
5.1 HGF-MEDIATED EGFR PARACRINE CROSSTALK 78
5.1.1 HGF IS A P O T E N T PARACRINE INDUCER O F EGFR LIGANDS 7 8
5.1.2 HGF-INDUCED EGFR NEGATIVE REGULATION 8 0
5.2 INTERACTION PATHWAYS BETWEEN BREAST CANCER CELLS AND MACROPHAGES IN
HUMAN AND MOUSE 82
IMAGE 5
6 S U M M A R Y 8 5
7 ZUSAMMENFASSUNG 8 6
8 REFERENCES 8 7
9 APPENDIX 9 6
9.1 ABBREVIATIONS 9 6
9.2 ACKNOWLEDGEMENTS 100
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| spelling | Gusenbauer, Simone Verfasser aut Macrophage-related interaction pathways in tumor stroma Simone Salome Gusenbauer 2012 100 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zsfassung in dt. Sprache München, Techn. Univ., Diss., 2012 Tumorassoziierter Makrophage (DE-588)4294570-7 gnd rswk-swf Protein-Tyrosin-Kinasen (DE-588)4409305-6 gnd rswk-swf Enzymaktivität (DE-588)4152470-6 gnd rswk-swf Signaltransduktion (DE-588)4318717-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Tumorassoziierter Makrophage (DE-588)4294570-7 s Signaltransduktion (DE-588)4318717-1 s Protein-Tyrosin-Kinasen (DE-588)4409305-6 s Enzymaktivität (DE-588)4152470-6 s b DE-604 Erscheint auch als Online-Ausgabe urn:nbn:de:bvb:91-diss-20120712-1094701-1-2 http://mediatum.ub.tum.de/node?id=1094701 Verlag kostenfrei Volltext https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120712-1094701-1-2 Resolving-System DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025214193&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Gusenbauer, Simone Macrophage-related interaction pathways in tumor stroma Tumorassoziierter Makrophage (DE-588)4294570-7 gnd Protein-Tyrosin-Kinasen (DE-588)4409305-6 gnd Enzymaktivität (DE-588)4152470-6 gnd Signaltransduktion (DE-588)4318717-1 gnd |
| subject_GND | (DE-588)4294570-7 (DE-588)4409305-6 (DE-588)4152470-6 (DE-588)4318717-1 (DE-588)4113937-9 |
| title | Macrophage-related interaction pathways in tumor stroma |
| title_auth | Macrophage-related interaction pathways in tumor stroma |
| title_exact_search | Macrophage-related interaction pathways in tumor stroma |
| title_full | Macrophage-related interaction pathways in tumor stroma Simone Salome Gusenbauer |
| title_fullStr | Macrophage-related interaction pathways in tumor stroma Simone Salome Gusenbauer |
| title_full_unstemmed | Macrophage-related interaction pathways in tumor stroma Simone Salome Gusenbauer |
| title_short | Macrophage-related interaction pathways in tumor stroma |
| title_sort | macrophage related interaction pathways in tumor stroma |
| topic | Tumorassoziierter Makrophage (DE-588)4294570-7 gnd Protein-Tyrosin-Kinasen (DE-588)4409305-6 gnd Enzymaktivität (DE-588)4152470-6 gnd Signaltransduktion (DE-588)4318717-1 gnd |
| topic_facet | Tumorassoziierter Makrophage Protein-Tyrosin-Kinasen Enzymaktivität Signaltransduktion Hochschulschrift |
| url | http://mediatum.ub.tum.de/node?id=1094701 https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120712-1094701-1-2 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025214193&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT gusenbauersimone macrophagerelatedinteractionpathwaysintumorstroma |