Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors:
Gespeichert in:
| Beteilige Person: | |
|---|---|
| Format: | Hochschulschrift/Dissertation Buch |
| Sprache: | Englisch |
| Veröffentlicht: |
2012
|
| Schlagwörter: | |
| Links: | http://mediatum.ub.tum.de/node?id=1107228 https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120821-1107228-0-4 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025699521&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| Umfang: | 113 Bl. Ill., graph. Darst. |
Internformat
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV040719258 | ||
| 003 | DE-604 | ||
| 005 | 20130322 | ||
| 007 | t| | ||
| 008 | 130201s2012 xx ad|| m||| 00||| eng d | ||
| 035 | |a (OCoLC)828790992 | ||
| 035 | |a (DE-599)BVBBV040719258 | ||
| 040 | |a DE-604 |b ger |e rakwb | ||
| 041 | 0 | |a eng | |
| 049 | |a DE-384 |a DE-473 |a DE-703 |a DE-1051 |a DE-824 |a DE-29 |a DE-12 |a DE-91 |a DE-19 |a DE-1049 |a DE-92 |a DE-739 |a DE-898 |a DE-355 |a DE-706 |a DE-20 |a DE-1102 | ||
| 100 | 1 | |a Grünewald, Thomas G. P. |d 1980- |e Verfasser |0 (DE-588)13673426X |4 aut | |
| 245 | 1 | 0 | |a Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors |c Thomas Georg Philipp Grünewald |
| 264 | 1 | |c 2012 | |
| 300 | |a 113 Bl. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a München, Techn. Univ., Diss., 2012 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 776 | 0 | 8 | |i Erscheint auch als |n Online-Ausgabe |o urn:nbn:de:bvb:91-diss-20120821-1107228-0-4 |
| 856 | 4 | 1 | |u http://mediatum.ub.tum.de/node?id=1107228 |x Verlag |z kostenfrei |3 Volltext |
| 856 | 4 | |u https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120821-1107228-0-4 |x Resolving-System | |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025699521&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 912 | |a ebook | ||
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-025699521 | |
Datensatz im Suchindex
| DE-BY-OTHR_katkey | 5072214 |
|---|---|
| DE-BY-TUM_call_number | 0001 DM 30360 |
| DE-BY-TUM_katkey | 1908825 |
| DE-BY-TUM_location | Mag |
| DE-BY-TUM_media_number | TEMP3217811 |
| _version_ | 1831256833025638400 |
| adam_text | IMAGE 1
1 INTRODUCTION
1
1.1 EWING TUMOR NOSOLOGY AND EPIDEMIOLOGY 1
1.2 CLINICAL AND BIOLOGICAL MARKERS FOR OUTCOME PREDICTION 3
1.3 TARGETED THERAPY OF EWING TUMORS 3
1.4 CURRENT STATE OF KNOWLEDGE ON STEAP1 4
1.5 THE DUAL ROLE OF REACTIVE OXYGEN SPECIES IN CANCER 7
2 RESEARCH OBJECTIVES AND SCIENTIFIC AIMS 10
2.1 RESEARCH OBJECTIVES 10
2.2 SCIENTIFIC AIMS 10
3 MATERIALS AND METHODS 11
3.1 MATERIALS 11
3.1.2 GENERAL MATERIALS 14
3.1.3 INSTRUMENTS AND EQUIPMENT 14
3.1.4 CHEMICAL AND BIOLOGICAL REAGENTS 16
3.1.5 COMMERCIAL REAGENT KITS 18
3.1.6 MEDIA, BUFFERS AND SOLUTIONS 19
3.1.6.1 CELL CULTURE MEDIA AND UNIVERSAL SOLUTIONS 19
3.1.6.2 BUFFERS AND GELS FOR WESTERN BLOT ANALYSIS 19
3.1.6.3 BUFFER AND GEL FOR DNA/RNA ELECTROPHORESIS 20
3.1.6.4 BUFFERS AND SOLUTIONS FOR FLOW CYTOMETRY AND CELL CYCLE ANALYSIS
20
3.1.7 HUMAN CELL LINES, MOUSE MODEL AND BACTERIAL STRAIN 20
3.1.7.1 HUMAN CANCER CELL LINES 21
3.1.7.2 MOUSE MODEL 21
3.1.7.3 BACTERIAL STRAIN USED FOR PLASMID EXPANSION 22
3.1.8 PRIMER ASSAYS USED FOR QUANTITATIVE REAL-TIME PCR (QRT-PCR) 2 2
3.1.9 SEQUENCES O F SMALL INTERFERING RNAS (SIRNAS) 23
3.1.10 SEQUENCES O F PRIMERS FOR CHROMATIN IMMUNOPRECIPITATION (CHIP) 23
3.1.11 ANTIBODIES FOR WESTERN BLOT AND IMMUNOHISTOCHEMISTRY 24
3.2 METHODS 2 5
3.2.1 CELL CULTURE CONDITIONS AND CELL CRYOCONSERVATION 25
3.2.2 TRANSFORMATION O F COMPETENT BACTERIA 2 6
3.2.3 MINI- AND MAXI-PREPARATION O F PLASMID DNA 2 6
3.2.4 AGAROSE GELELECTROPHORESIS 2 6
3.2.5 ANNEALING O F OLIGONUCLEOTIDES 2 7
3.2.6 RNA ISOLATION USING RNEASY MINI KIT 2 7
3.2.7 ISOLATION O F TOTAL RNA USING TRI REAGENT RNA ISOLATION KIT 2 7
HTTP://D-NB.INFO/1036941965
IMAGE 2
3.2.8 REVERSE TRANSCRIPTION 28
3.2.9 QUANTITATIVE REAL-TIME PCR (QRT-PCR) 28
3.2.10 DETECTION O F EWS/FLI1 ; 29
3.2.11 LIGATION O F DNA FRAGMENTS, CONSTRUCTS AND RETROVIRAL GENE
TRANSFER 29
3.2.12 CHROMATIN IMMUNOPRECIPITATION (CHIP) 30
3.2.13 WESTERN BLOT (WB) 31
3.2.14 RNA INTERFERENCE (RNAI) 31
3.2.15 MICROARRAYS 3 2
3.2.16 MEASUREMENT O F ROS AND MITOCHONDRIAL MASS 33
3.2.17 FLOW CYTOMETRY 34
3.2.18 ASSESSMENT O F CELL CYCLE, APOPTOSIS AND NECROSIS 35
3.2.19 PROLIFERATION ASSAY 35
3.2.20 INVASION ASSAY 35
3.2.21 COLONY FORMING ASSAY 36
3.2.22 ANGIOGENESIS ASSAY 36
3.2.23 MIGRATION ASSAY 36
3.2.24 INDIRECT IMMUNOFLUORESCENCE (IF) 3 7
3.2.25 ADHESION ASSAY 3 7
3.2.26 MICE AND IN VIVO EXPERIMENTS 3 7
3.2.27 GLUTATHIONE ASSAY 38
3.2.28 ELECTRON MICROSCOPY 38
3.2.29 2D GELELECTROPHORESIS, SPOT SELECTION AND MASS SPECTROMETRY 39
3.2.30 STUDY POPULATION, E T SAMPLES AND TISSUE MICROARRAY (TMA) 40
3.2.31 IMMUNOHISTOCHEMISTRY (IHC) AND EVALUATION O F IMMUNOREACTIVITY 41
3.2.32 ANALYSIS O F CELLULAR IRON CONTENT 43
3.2.33 ELI SPOT ANALYSIS FOR DETECTION O F INTERFERON SECRETION 43
3.2.34 STATISTICAL ANALYSES 43
4 RESULTS 45
4.1 ANALYSIS OF STEAP1 IN VITRO AND IN E T XENOGRAFTS 4 5
4.1.1 STEAP1 IS INDUCED BY EWS/FLI1 AND HIGHLY EXPRESSED IN E T 45
4.1.2 SILENCING OFSTEAPI NEITHER AFFECTS NEUROENDOTHELIAL PHENOTYPE NOR
ALTERS
CELLULAR MIGRATION AND ADHESION IN VITRO 47
4.1.3 KNOCKDOWN OFSTEAPI INHIBITS PROLIFERATION, INVASION,
ANCHORAGEINDEPENDENT COLONY-FORMATION, TUMORIGENICITY AND METASTASIS O F
E T CELLS 47
IMAGE 3
4.1.4 STEAP1 EXPRESSION NEITHER IMPACTS CELL SURVIVAL NOR CELL CYCLE
PROGRESSION,
BUT CORRELATES WITH INVASIVENESS 51
4.1.5 STEAP1 -SILENCING LEADS TO ADAPTATIONS IN OXIDATIVE STRESS
RESPONSE
SYSTEMS 5 2
4.1.6 STEAP1 EXPRESSION IS NOT LINKED WITH IRON UPTAKE AND REDUCTION,
BUT
INFLUENCES KEY COMPONENTS O F THE UPS 55
4.1.7 STEAP1-SILENCING REGULATES REDOX-SENSITIVE MIRNAS 57
4.1.8 STEAP1 EXPRESSION IS ASSOCIATED WITH ROS LEVELS O F E T CELLS 58
4.1.9 ROS ARE CRITICAL FOR E T PROLIFERATION AND INVASIVENESS 59
4.1.10 STEAP1 KNOCKDOWN INHIBITS STAT1 ACTIVATION 62
4.2 ANALYSIS OF STEAP1 IN CLINICAL E T SAMPLES 6 4
4.2.1 STEAP1 IS EXPRESSED IN MOST CLINICAL E T AND PREDOMINANTLY LOCATES
TO
PLASMA MEMBRANES 64
4.2.2 MEMBRANOUS AND CYTOPLASMIC STEAP1 EXPRESSION AND OVERALL SURVIVAL_
65
4.2.3 MULTIVARIATE ANALYSIS 66
4.2.4 ASSOCIATION O F STEAP1 EXPRESSION WITH HISTOLOGICAL RESPONSE TO
CHEMOTHERAPY 68
5 DISCUSSION 70
6 CONCLUSIONS, LIMITATIONS AND PERSPECTIVES 77
6.1 CONCLUSIONS 7 7
6.2 LIMITATIONS AND PERSPECTIVES 7 7
7 SUMMARY 80
8 REFERENCES 82
9 APPENDIX 95
9.1 LIST OF DIFFERENTIALLY REGULATED GENES AS IDENTIFIED BY MICROARRAYS
9 5
9.2 LIST OF DIFFERENTIALLY REGULATED PROTEINS AS IDENTIFIED BY MASS
SPECTROMETRY 9 8
9.3 LIST OF DIFFERENTIALLY REGULATED MIRNAS 102
9.4 CLINICOPATHOLOGICAL CHARACTERISTICS OF PATIENTS INCLUDED IN T M A
105
9.5 LIST OF FIGURES 1 0 9
9.6 LIST OF TABLES 1 1 0
9.7 LIST OF ABBREVIATIONS 111
10 ACKNOWLEDGEMENT 112
11 PUBLICATIONS DERIVED FROM THIS PH.D. THESIS 113
|
| any_adam_object | 1 |
| author | Grünewald, Thomas G. P. 1980- |
| author_GND | (DE-588)13673426X |
| author_facet | Grünewald, Thomas G. P. 1980- |
| author_role | aut |
| author_sort | Grünewald, Thomas G. P. 1980- |
| author_variant | t g p g tgp tgpg |
| building | Verbundindex |
| bvnumber | BV040719258 |
| collection | ebook |
| ctrlnum | (OCoLC)828790992 (DE-599)BVBBV040719258 |
| format | Thesis Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01591nam a2200325 c 4500</leader><controlfield tag="001">BV040719258</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">20130322 </controlfield><controlfield tag="007">t|</controlfield><controlfield tag="008">130201s2012 xx ad|| m||| 00||| eng d</controlfield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)828790992</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)BVBBV040719258</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-384</subfield><subfield code="a">DE-473</subfield><subfield code="a">DE-703</subfield><subfield code="a">DE-1051</subfield><subfield code="a">DE-824</subfield><subfield code="a">DE-29</subfield><subfield code="a">DE-12</subfield><subfield code="a">DE-91</subfield><subfield code="a">DE-19</subfield><subfield code="a">DE-1049</subfield><subfield code="a">DE-92</subfield><subfield code="a">DE-739</subfield><subfield code="a">DE-898</subfield><subfield code="a">DE-355</subfield><subfield code="a">DE-706</subfield><subfield code="a">DE-20</subfield><subfield code="a">DE-1102</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Grünewald, Thomas G. P.</subfield><subfield code="d">1980-</subfield><subfield code="e">Verfasser</subfield><subfield code="0">(DE-588)13673426X</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors</subfield><subfield code="c">Thomas Georg Philipp Grünewald</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2012</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">113 Bl.</subfield><subfield code="b">Ill., graph. Darst.</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="a">München, Techn. Univ., Diss., 2012</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="776" ind1="0" ind2="8"><subfield code="i">Erscheint auch als</subfield><subfield code="n">Online-Ausgabe</subfield><subfield code="o">urn:nbn:de:bvb:91-diss-20120821-1107228-0-4</subfield></datafield><datafield tag="856" ind1="4" ind2="1"><subfield code="u">http://mediatum.ub.tum.de/node?id=1107228</subfield><subfield code="x">Verlag</subfield><subfield code="z">kostenfrei</subfield><subfield code="3">Volltext</subfield></datafield><datafield tag="856" ind1="4" ind2=" "><subfield code="u">https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120821-1107228-0-4</subfield><subfield code="x">Resolving-System</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025699521&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">ebook</subfield></datafield><datafield tag="943" ind1="1" ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-025699521</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV040719258 |
| illustrated | Illustrated |
| indexdate | 2024-12-20T16:24:08Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-025699521 |
| oclc_num | 828790992 |
| open_access_boolean | 1 |
| owner | DE-384 DE-473 DE-BY-UBG DE-703 DE-1051 DE-824 DE-29 DE-12 DE-91 DE-BY-TUM DE-19 DE-BY-UBM DE-1049 DE-92 DE-739 DE-898 DE-BY-UBR DE-355 DE-BY-UBR DE-706 DE-20 DE-1102 |
| owner_facet | DE-384 DE-473 DE-BY-UBG DE-703 DE-1051 DE-824 DE-29 DE-12 DE-91 DE-BY-TUM DE-19 DE-BY-UBM DE-1049 DE-92 DE-739 DE-898 DE-BY-UBR DE-355 DE-BY-UBR DE-706 DE-20 DE-1102 |
| physical | 113 Bl. Ill., graph. Darst. |
| psigel | ebook |
| publishDate | 2012 |
| publishDateSearch | 2012 |
| publishDateSort | 2012 |
| record_format | marc |
| spellingShingle | Grünewald, Thomas G. P. 1980- Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors |
| subject_GND | (DE-588)4113937-9 |
| title | Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors |
| title_auth | Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors |
| title_exact_search | Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors |
| title_full | Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors Thomas Georg Philipp Grünewald |
| title_fullStr | Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors Thomas Georg Philipp Grünewald |
| title_full_unstemmed | Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors Thomas Georg Philipp Grünewald |
| title_short | Functional characterization of the protein STEAP1 for the development of specific biomarkers and targeted therapeutics for Ewing tumors |
| title_sort | functional characterization of the protein steap1 for the development of specific biomarkers and targeted therapeutics for ewing tumors |
| topic_facet | Hochschulschrift |
| url | http://mediatum.ub.tum.de/node?id=1107228 https://nbn-resolving.org/urn:nbn:de:bvb:91-diss-20120821-1107228-0-4 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025699521&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT grunewaldthomasgp functionalcharacterizationoftheproteinsteap1forthedevelopmentofspecificbiomarkersandtargetedtherapeuticsforewingtumors |
Online lesen (frei zugänglich)
Inhaltsverzeichnis
Paper/Kapitel scannen lassen
Inhaltsverzeichnis
Paper/Kapitel scannen lassen
Bibliotheksmagazin
| Signatur: |
0001 DM 30360
Lageplan |
|---|---|
| Exemplar 1 | Ausleihbar Am Standort |