Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma:
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| Main Author: | |
|---|---|
| Format: | Book |
| Language: | English |
| Published: |
Köln
[2021]
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Item Description: | Enthält Sonderabdruck aus Zeitschrift. - Zusammenfassung in deutscher und englischer Sprache |
| Physical Description: | X, 176, XIII, Seiten Illustrationen, Diagramme |
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| 245 | 1 | 0 | |a Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma |c vorgelegt von Nasrin Refaian aus Köln |
| 264 | 1 | |a Köln |c [2021] | |
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| adam_text | CONTENTS
CONTENTS
...............................................................................................................................
I
LIST
OF
FIGURES
....................................................................................................................IV
LIST
OF
TABLES
.....................................................................................................................
VII
CODE
OF
ABBREVIATIONS
...................................................................................................VIII
ZUSAMMENFASSUNG
............................................................................................................
1
ABSTRACT
...............................................................................................................................
3
1.
INTRODUCTION
..................................................................................................................
5
1.1
CANCER
...................................................................................................................
5
1.2
MALIGNANT
MELANOMA
............................................................................................
6
1.2.1
UVEAL
MELANOMA
........................................................................................
6
1.2.2
CONJUNCTIVAL
MELANOMA
.............................................................................
8
1.3
METASTASIS
OF
OCULAR
MELANOMA
.........................................................................
10
1.4
TUMOR
MICROENVIRONMENT
IN
OCULAR
MELANOMA
.................................................
13
1.4.1
ENDOTHELIAL
CELLS
.......................................................................................
13
1.4.2
CANCER-ASSOCIATED
FIBROBLASTS
.................................................................
14
1.5
AIMS
....................................................................................................................
17
2.
MATERIAL
........................................................................................................................
19
2.1
CHEMICALS,
REAGENTS,
AND
SOLVENTS
....................................................................
19
2.2
BUFFERS
AND
SOLUTIONS
..........................................................................................
20
2.3
PROTEINS
...............................................................................................................
21
2.4
ANTIBODIES,
DYES,
AND
ISOTYPE
CONTROLS
..............................................................
21
2.5
COMMERCIALLY
AVAILABLE
KITS
...............................................................................
23
2.6
CELL
CULTURE
MEDIA
AND
SUPPLEMENTS
.................................................................
23
2.7
CELLS
AND
CELL
LINES
..............................................................................................
27
2.8
CONSUMABLES
.....................................................................................................
27
2.9
EQUIPMENT
...........................................................................................................
29
2.10
SOFTWARE
..............................................................................................................
30
3.
METHODS
......................................................................................................................
32
3.1
CELL
CULTURE
.....................
32
3.1.1
CULTURING
CONDITIONS
.................................................................................
32
3.1.2
ISOLATION
OF
FIBROBLASTS
.............................................................................
32
3.1.3
DETERMINATION
OF
CELL
COUNT
.....................................................................
33
3.1.4
CONDITIONED
MEDIA
...................................................................................
33
3.2
CHARACTERIZATION
OF
FIBROBLASTS
...........................................................................
34
3.2.1
IMMUNOCYTOCHEMISTRY
............................................................................
34
II
CONTENTS
3.2.2
FLOW
CYTOMETRY
.......................................................................................
35
3.3
PROTEOMICS
...........................................................................................................
37
3.3.1
STABLE
ISOTOPE
LABELING
BY
AMINO
ACIDS
IN
CELL
CULTURE
...........................
37
3.3.2
PHOSPHOPROTEOMICS
.................................................................................
42
3.3.3
ENZYME-LINKED
IMMUNOSORBENT
ASSAY
(ELISA)
.....................................
45
3.4
FUNCTIONAL
ASSAYS
...............................................................................................
46
3.4.1
CELL
VIABILITY
ASSAY
...................................................................................
46
3.4.2
CELL
PROLIFERATION
ASSAY
............................................................................
47
3.4.3
CELL
MIGRATION
(SCRATCH
WOUND
HEALING)
ASSAY
.....................................
48
3.4.4
ENDOTHELIAL
TUBE
FORMATION
ASSAY
...........................................................
51
3.5
STATISTICAL
ANALYSES
.............................................................................................
52
4.
RESULTS
.........................................................................................................................
54
4.1
CHARACTERIZATION
OF
FIBROBLASTS
...........................................................................
54
4.1.1
CHARACTERIZATION
OF
HCMFS
AND
HCFS
....................................................
54
4.1.2
VALIDATION
OF
MARKER
EXPRESSION
OF
HCMFS
AND
HCFS
...........................
62
4.2
PROTEOMICS
...........................................................................................................
63
4.2.1
DETERMINING
HCMF
AND
HCF
VIABILITY
UNDER
SILAC
LABELING
CONDITIONS
.................................................................................................
64
4.2.2
PROTEOME
AND
SECRETOME
PROFILE
OF
HCMFS
AND
HCFS
USING
SILAC
.
.
64
4.2.3
DETERMINING
CMC
AND
MC
VIABILITY
UNDER
SILAC
LABELING
CONDITIONS
.................................................................................................
68
4.2.4
PROTEOME
AND
SECRETOME
PROFILING
OF
MCS
AND
CMCS
USING
SILAC
....69
4.2.5
PHOSPHOPROTEOMICS
OF
CMCS
AND
MCS
.................................................
78
4.2.6
QUANTITATIVE
ANALYSIS
OF
SECRETED
PRO(LYMPH-)ANGIOGENIC
PROTEINS
.......
93
4.3
FUNCTIONAL
ASSAYS
...............................................................................................
95
4.3.1
EFFECT
OF
SECRETED
PROTEINS
ON
ENDOTHELIAL
CELL
PROLIFERATION
..................
95
4.3.2
EFFECT
OF
SECRETED
PROTEINS
ON
MELANOMA
CELL
PROLIFERATION
...................
97
4.3.3
INFLUENCE
OF
SFLT1
ON
EC
AND
CMC
PROLIFERATION
..................................
99
4.3.4
EFFECT
OF
SECRETED
PROTEINS
ON
ENDOTHELIAL
CELL
MIGRATION
.....................
100
4.3.5
EFFECT
OF
SECRETED
PROTEINS
ON
THE
DEVELOPMENT
OF
CAPILLARY-LIKE
STRUCTURES
OF
ECS
....................................................................................
109
5.
DISCUSSION
................................................................................................................
114
5.1
HETEROGENEITY
OF
TME
.......................................................................................
114
5.2
QUANTITATIVE
PROTEOMICS
OF
HCMFS
AND
CMCS
...............................................
116
5.3
CMCS
INDUCE
TUMOR-ASSOCIATED
(LYMPH-)ANGIOGENESIS
AND
METASTASIS
......
118
5.4
INFLUENCE
OF
TME
ON
(LYMPH-)ANGIOGENESIS
AND
METASTASIS
.........................
121
5.5
INFLUENCE
OF
CFS
AND
CMCS
ON
MELANOMA
GROWTH
........................................
123
5.6
ACTIVATED
SIGNALING
PATHWAYS
IN
CMS
...............................................................
124
5.7
CONCLUSION
AND
OUTLOOK
....................................................................................
132
SUPPLEMENT
A:
DATA
.......................................................................................................
136
A.1
CHARACTERIZATION
OF
FIBROBLASTS
(IN
ADDITION
TO
CHAPTER
4.1)
.............................
136
CONTENTS
III
A.
2
PROLIFERATION
OF
ENDOTHELIAL
CELLS
(IN
ADDITION
TO
CHAPTER
4.3.1)
........................
139
SUPPLEMENT
B:
PUBLICATIONS
........................................................................................
140
B.
1
COMPARING
THE
HEM
AND
LYMPHANGIOGENIC
PROFILE
OF
CONJUNCTIVAL
AND
UVEAL
MELANOMA
CELL
LINES
.............................................................................
140
B.2
AUTOCRINE
IMPACT
OF
VEGF-A
ON
UVEAL
MELANOMA
CELLS
.................................
147
REFERENCES
......................................................................................................................
155
ACKNOWLEDGMENT
.............................................................................................................
XI
ERKLARUNG
.........................................................................................................................
XIII
|
| adam_txt |
CONTENTS
CONTENTS
.
I
LIST
OF
FIGURES
.IV
LIST
OF
TABLES
.
VII
CODE
OF
ABBREVIATIONS
.VIII
ZUSAMMENFASSUNG
.
1
ABSTRACT
.
3
1.
INTRODUCTION
.
5
1.1
CANCER
.
5
1.2
MALIGNANT
MELANOMA
.
6
1.2.1
UVEAL
MELANOMA
.
6
1.2.2
CONJUNCTIVAL
MELANOMA
.
8
1.3
METASTASIS
OF
OCULAR
MELANOMA
.
10
1.4
TUMOR
MICROENVIRONMENT
IN
OCULAR
MELANOMA
.
13
1.4.1
ENDOTHELIAL
CELLS
.
13
1.4.2
CANCER-ASSOCIATED
FIBROBLASTS
.
14
1.5
AIMS
.
17
2.
MATERIAL
.
19
2.1
CHEMICALS,
REAGENTS,
AND
SOLVENTS
.
19
2.2
BUFFERS
AND
SOLUTIONS
.
20
2.3
PROTEINS
.
21
2.4
ANTIBODIES,
DYES,
AND
ISOTYPE
CONTROLS
.
21
2.5
COMMERCIALLY
AVAILABLE
KITS
.
23
2.6
CELL
CULTURE
MEDIA
AND
SUPPLEMENTS
.
23
2.7
CELLS
AND
CELL
LINES
.
27
2.8
CONSUMABLES
.
27
2.9
EQUIPMENT
.
29
2.10
SOFTWARE
.
30
3.
METHODS
.
32
3.1
CELL
CULTURE
.
32
3.1.1
CULTURING
CONDITIONS
.
32
3.1.2
ISOLATION
OF
FIBROBLASTS
.
32
3.1.3
DETERMINATION
OF
CELL
COUNT
.
33
3.1.4
CONDITIONED
MEDIA
.
33
3.2
CHARACTERIZATION
OF
FIBROBLASTS
.
34
3.2.1
IMMUNOCYTOCHEMISTRY
.
34
II
CONTENTS
3.2.2
FLOW
CYTOMETRY
.
35
3.3
PROTEOMICS
.
37
3.3.1
STABLE
ISOTOPE
LABELING
BY
AMINO
ACIDS
IN
CELL
CULTURE
.
37
3.3.2
PHOSPHOPROTEOMICS
.
42
3.3.3
ENZYME-LINKED
IMMUNOSORBENT
ASSAY
(ELISA)
.
45
3.4
FUNCTIONAL
ASSAYS
.
46
3.4.1
CELL
VIABILITY
ASSAY
.
46
3.4.2
CELL
PROLIFERATION
ASSAY
.
47
3.4.3
CELL
MIGRATION
(SCRATCH
WOUND
HEALING)
ASSAY
.
48
3.4.4
ENDOTHELIAL
TUBE
FORMATION
ASSAY
.
51
3.5
STATISTICAL
ANALYSES
.
52
4.
RESULTS
.
54
4.1
CHARACTERIZATION
OF
FIBROBLASTS
.
54
4.1.1
CHARACTERIZATION
OF
HCMFS
AND
HCFS
.
54
4.1.2
VALIDATION
OF
MARKER
EXPRESSION
OF
HCMFS
AND
HCFS
.
62
4.2
PROTEOMICS
.
63
4.2.1
DETERMINING
HCMF
AND
HCF
VIABILITY
UNDER
SILAC
LABELING
CONDITIONS
.
64
4.2.2
PROTEOME
AND
SECRETOME
PROFILE
OF
HCMFS
AND
HCFS
USING
SILAC
.
.
64
4.2.3
DETERMINING
CMC
AND
MC
VIABILITY
UNDER
SILAC
LABELING
CONDITIONS
.
68
4.2.4
PROTEOME
AND
SECRETOME
PROFILING
OF
MCS
AND
CMCS
USING
SILAC
.69
4.2.5
PHOSPHOPROTEOMICS
OF
CMCS
AND
MCS
.
78
4.2.6
QUANTITATIVE
ANALYSIS
OF
SECRETED
PRO(LYMPH-)ANGIOGENIC
PROTEINS
.
93
4.3
FUNCTIONAL
ASSAYS
.
95
4.3.1
EFFECT
OF
SECRETED
PROTEINS
ON
ENDOTHELIAL
CELL
PROLIFERATION
.
95
4.3.2
EFFECT
OF
SECRETED
PROTEINS
ON
MELANOMA
CELL
PROLIFERATION
.
97
4.3.3
INFLUENCE
OF
SFLT1
ON
EC
AND
CMC
PROLIFERATION
.
99
4.3.4
EFFECT
OF
SECRETED
PROTEINS
ON
ENDOTHELIAL
CELL
MIGRATION
.
100
4.3.5
EFFECT
OF
SECRETED
PROTEINS
ON
THE
DEVELOPMENT
OF
CAPILLARY-LIKE
STRUCTURES
OF
ECS
.
109
5.
DISCUSSION
.
114
5.1
HETEROGENEITY
OF
TME
.
114
5.2
QUANTITATIVE
PROTEOMICS
OF
HCMFS
AND
CMCS
.
116
5.3
CMCS
INDUCE
TUMOR-ASSOCIATED
(LYMPH-)ANGIOGENESIS
AND
METASTASIS
.
118
5.4
INFLUENCE
OF
TME
ON
(LYMPH-)ANGIOGENESIS
AND
METASTASIS
.
121
5.5
INFLUENCE
OF
CFS
AND
CMCS
ON
MELANOMA
GROWTH
.
123
5.6
ACTIVATED
SIGNALING
PATHWAYS
IN
CMS
.
124
5.7
CONCLUSION
AND
OUTLOOK
.
132
SUPPLEMENT
A:
DATA
.
136
A.1
CHARACTERIZATION
OF
FIBROBLASTS
(IN
ADDITION
TO
CHAPTER
4.1)
.
136
CONTENTS
III
A.
2
PROLIFERATION
OF
ENDOTHELIAL
CELLS
(IN
ADDITION
TO
CHAPTER
4.3.1)
.
139
SUPPLEMENT
B:
PUBLICATIONS
.
140
B.
1
COMPARING
THE
HEM
AND
LYMPHANGIOGENIC
PROFILE
OF
CONJUNCTIVAL
AND
UVEAL
MELANOMA
CELL
LINES
.
140
B.2
AUTOCRINE
IMPACT
OF
VEGF-A
ON
UVEAL
MELANOMA
CELLS
.
147
REFERENCES
.
155
ACKNOWLEDGMENT
.
XI
ERKLARUNG
.
XIII |
| any_adam_object | 1 |
| any_adam_object_boolean | 1 |
| author | Refaian, Nasrin |
| author_GND | (DE-588)1255739460 |
| author_facet | Refaian, Nasrin |
| author_role | aut |
| author_sort | Refaian, Nasrin |
| author_variant | n r nr |
| building | Verbundindex |
| bvnumber | BV048285336 |
| ctrlnum | (DE-599)DNB1255735546 |
| discipline | Medizin |
| discipline_str_mv | Medizin |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
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| id | DE-604.BV048285336 |
| illustrated | Illustrated |
| index_date | 2024-07-03T20:01:50Z |
| indexdate | 2024-07-10T09:34:11Z |
| institution | BVB |
| language | English |
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| physical | X, 176, XIII, Seiten Illustrationen, Diagramme |
| publishDate | 2021 |
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| spelling | Refaian, Nasrin Verfasser (DE-588)1255739460 aut Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma vorgelegt von Nasrin Refaian aus Köln Köln [2021] X, 176, XIII, Seiten Illustrationen, Diagramme txt rdacontent n rdamedia nc rdacarrier Enthält Sonderabdruck aus Zeitschrift. - Zusammenfassung in deutscher und englischer Sprache (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1255735546/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=033665459&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Refaian, Nasrin Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma |
| subject_GND | (DE-588)4113937-9 |
| title | Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma |
| title_auth | Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma |
| title_exact_search | Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma |
| title_exact_search_txtP | Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma |
| title_full | Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma vorgelegt von Nasrin Refaian aus Köln |
| title_fullStr | Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma vorgelegt von Nasrin Refaian aus Köln |
| title_full_unstemmed | Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma vorgelegt von Nasrin Refaian aus Köln |
| title_short | Influence of melanoma cells and cancer-associated fibroblasts on (lymph-)angiogenesis in conjunctival melanoma |
| title_sort | influence of melanoma cells and cancer associated fibroblasts on lymph angiogenesis in conjunctival melanoma |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1255735546/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=033665459&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT refaiannasrin influenceofmelanomacellsandcancerassociatedfibroblastsonlymphangiogenesisinconjunctivalmelanoma |
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