The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway: microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line
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| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2004
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | VII, 132 Bl. graph. Darst. 30 cm |
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MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV019774551 | ||
| 003 | DE-604 | ||
| 005 | 20050620 | ||
| 007 | t | ||
| 008 | 050415s2004 gw d||| m||| 00||| eng d | ||
| 015 | |a 04,H11,2347 |2 dnb | ||
| 016 | 7 | |a 971726337 |2 DE-101 | |
| 035 | |a (DE-599)BVBBV019774551 | ||
| 040 | |a DE-604 |b ger |e rakddb | ||
| 041 | 0 | |a eng | |
| 044 | |a gw |c XA-DE | ||
| 084 | |a 610 |2 sdnb | ||
| 100 | 1 | |a Wu, Xiaoqing |d 1966- |e Verfasser |0 (DE-588)129191345 |4 aut | |
| 245 | 1 | 0 | |a The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway |b microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line |c von Xiaoqing Wu |
| 264 | 1 | |c 2004 | |
| 300 | |a VII, 132 Bl. |b graph. Darst. |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Berlin, Humboldt-Univ., Diss., 2004 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 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=013100635&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-013100635 | |
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| _version_ | 1807503904128106496 |
|---|---|
| adam_text |
CONTENTS
1.
INTRODUCTION
.
1
1.1.
SMAD4
-
A
POTENT
TUMOR
SUPPRESSOR
.
2
1.2.
TGF-B
-
A
SUPERFAMILY
OF
CYTOKINES
.
4
1.2.1.
TGF-B
EXPRESSION
AND
ITS
ISOFORMS
.
4
1.2.2.
TGF-B
SECRETION
AND
ACTIVATION
.
5
1.2.3.
MULTIPLEX
FUNCTIONS
OF
TGF-B
.
5
1.2.4.
THE
DUAL
ROLE
OF
TGF-B
IN
TUMORIGENESIS
.
7
1.3.
THE
ROLE
OF
THE
SMAD4
PROTEINS
IN
TGF-B
SIGNALING
.
8
1.3.1.
THE
TGF-B
RECEPTOR
FAMILY
.
9
1.3.2.
THE
SMAD
FAMILY
.
9
1.3.3.
THE
TGF-B
SIGNALING
PATHWAY
.
11
1.3.4.
CROSSTALK
OF
THE
TGF-B
SIGNALING
PATHWAY
AND
THE
RAS/MAPK
PATHWAY
.
14
1.4.
TRANSCRIPTIONAL
ACTIVITIES
REGULATED
BY
TGF-B
AND
SMADS
.
17
2.
AIMS
.
20
3.
MATERIALS
AND
METHODS
.
21
3.1.
MATERIALS
.
21
3.1.1.
APPLIANCES
.
21
3.1.2.
CHEMICALS,
SOLUTIONS
AND
BUFFERS
.
21
3.1.3.
PLASMIDS
AND
VIRUSES
.
23
3.1.4.
ENZYMES
.
24
3.1.5.
KITS
.
25
3.1.6.
ANTIBODIES
.
26
3.1.7.
OLIGONUCLEOTIDES
.
27
3.1.8.
SOFTWARE
.
28
3.2.
METHODS
.
28
3.2.1.
CELL
LINES
AND
CELL
CULTURE
.
28
3.2.2.
STABLE
T
RANSFECTION
.
28
3.2.3.
IN
SITU
X-GAL
STAINING
.
29
3.2.4.
WESTERN
BLOT
ANALYSIS
.
29
3.2.5.
TOTAL
RNA
PREPARATION
.
30
3.2.6.
CDNA
SYNTHESIS
.
31
3.2.7.
REVERSE
TRANSCRIPTASE
PCR
(RT-PCR)
.
31
3.2.8.
TRANSIENT
TRANSFECTIONS
AND
LUCIFERASE
REPORTER
ASSAY
.
32
3.2.9.
CELL
CYCLE
ANALYSIS
.
32
3.2.10.
IN
VITRO
CELL
GROWTH
ASSAY
.
33
3.2.11.
OLIGONUCLEOTIDE
MICROARRAY
ANALYSIS
.
33
3.2.12.
RELATIVE
QUANTITATIVE
RT-PCR
.
36
3.2.13.
ADENOVIRAL
INFECTION
.
37
3.2.14.
IMMUNOPRECIPITATION
.
38
3.2.15.
ISOLATION
OF
PLASMID
DNA
FROM
E.
COLI
.
.
38
3.2.16. INVASION
ASSAY
.
38
3.2.17.
MEASUREMENT
OF
TGF-B1
BY
ELISA
.
39
4.
RESULTS
.
40
4.1.
RECONSTITUTION
OF
FUNCTIONAL
SMAD4
IN
THE
MDA-MB-468
BREAST
CARCINOMA
CELL
LINE
.
40
4.1.1.
STABLE
REEXPRESSION
OF
SMAD4
.
40
4.1.2.
RESTORATION
OF
TGF-B-RESPONSIVENESS
IN
MDA-MB-468
STABLE
CLONES
.
42
4.1.2.1.
SMAD4
RESTORES
TGF-B-MEDIATED
INDUCTION
OF
P21
WAF1
.
42
4.1.2.2.
TGF-B
INDUCED
G1-ARREST
IN
MDA-MB-468
CELLS
REQUIRES
THE
PRESENCE
OF
SMAD4
.
45
4.1.2.3.
TGF-B
INDUCED
INHIBITION
OF
CELL
PROLIFERATION
IS
SMAD4-DEPENDENT
.
46
4.1.2.4.
TGF-B
INDUCES
THE
ACTIVATION
OF
THE
PAI-1
PROMOTER
IN
THE
PRESENCE
OF
SMAD4
.
47
4.2.
GENE
EXPRESSION
PROFILING
USING
AFFYMETRIX
OLIGONUCLEOTIDE
MICROARRAYS
.
49
4.2.1.
DATA
ANALYSIS
.
49
4.2.1.1.
ABSOLUTE
ANALYSIS
.
51
4.2.1.2.
COMPARISON
ANALYSIS
.
54
4.2.1.3.
GENE
SORTING
ACCORDING
TO
EXPRESSION
PROFILES
.
57
4.2.1.4.
VALIDATION
OF
ARRAY
DATA
.
61
4.2.1.5.
FUNCTIONAL
CLASSIFICATION
OF
SMAD4-DEPENDENT
TGF-B-REGULATED
GENE
63
4.2.1.5.1.
FUNCTIONAL
CLASSIFICATION
OF
SMAD4-DEPENDENT
INDUCED
GENES
.
63
4.2.1.5.2.
FUNCTIONAL
CLASSIFICATION
OF
SUPPRESSED
GENES
.
69
4.2.2. DATA
CONFIRMATION
OF
SELECTED
GENES
BY
QUANTITATIVE
RT-PCR
.
71
4.3.
INVESTIGATION
OF
CELL
INVASION
IN
VITRO
.
76
4.4.
TGF-B
INDUCES
THE
EXPRESSION
OF
E-CADHERIN
IN
THE
PRESENCE
OF
SMAD4
.
80
VI
4.5.
INVESTIGATION
OF
SIGNALING
PATHWAYS
RESPONSIBLE
FOR
SMAD4-
DEPENDENT
GENE
REGULATION
IN
THE
ABSENCE
OF
EXOGENOUS
TGF-B
.
81
4.5.1.
MEASUREMENT
OF
ACTIVE
TGF-B
IN
CELL
CULTURE
SUPERNATANTS
.
82
4.5.2.
ANALYSIS
OF
SIGNALING
VIA
R-SMADS
IN
THE
ABSENCE
OF
EXOGENOUS
TGF-B
.
83
4.5.3.
DETERMINATION
OF
MAPK
ACTIVITIES
IN
THE
ABSENCE
OF
EXOGENOUS
TGF-B
.
84
5.
DISCUSSION
.
95
5.1.
THE
INCREASING
COMPLEXITY
OF
SMAD4
FUNCTION
.
95
5.2.
SMAD4-RECONSTITUTED
CELLS
REGAIN
TGF-B
RESPONSIVENESS
.
97
5.3.
INTERPRETATION
OF
ARRAY
DATA
WITH
REGARD
TO
THE
DUAL
ROLE
OF
SMAD4
AND
TGF-B
IN
TUMORIGENESIS
.
99
5.4.
THE
ROLE
OF
SMAD4
IN
CANCER
CELL
INVASION
.
104
5.5.
THE
ROLE
OF
SERPINS
IN
SMAD4
'
S
TUMOR
SUPPRESSOR
FUNCTION
.
107
6.
CONCLUDING
REMARKS
.
111
7.
LITERATURE
.
112
8.
ABBREVIATION
.
128
9.
APPENDIX
.
129
9.1.
CURRICULUM
VITAE
.
129
9.2.
ACKNOWLEDGEMENTS
.
131
9.3.
STATEMENT
.
132
VII |
| any_adam_object | 1 |
| author | Wu, Xiaoqing 1966- |
| author_GND | (DE-588)129191345 |
| author_facet | Wu, Xiaoqing 1966- |
| author_role | aut |
| author_sort | Wu, Xiaoqing 1966- |
| author_variant | x w xw |
| building | Verbundindex |
| bvnumber | BV019774551 |
| ctrlnum | (DE-599)BVBBV019774551 |
| discipline | Medizin |
| format | Thesis Book |
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| genre_facet | Hochschulschrift |
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| indexdate | 2024-08-16T01:00:49Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-013100635 |
| open_access_boolean | |
| physical | VII, 132 Bl. graph. Darst. 30 cm |
| publishDate | 2004 |
| publishDateSearch | 2004 |
| publishDateSort | 2004 |
| record_format | marc |
| spelling | Wu, Xiaoqing 1966- Verfasser (DE-588)129191345 aut The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line von Xiaoqing Wu 2004 VII, 132 Bl. graph. Darst. 30 cm txt rdacontent n rdamedia nc rdacarrier Berlin, Humboldt-Univ., Diss., 2004 (DE-588)4113937-9 Hochschulschrift gnd-content DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=013100635&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Wu, Xiaoqing 1966- The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line |
| subject_GND | (DE-588)4113937-9 |
| title | The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line |
| title_auth | The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line |
| title_exact_search | The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line |
| title_full | The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line von Xiaoqing Wu |
| title_fullStr | The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line von Xiaoqing Wu |
| title_full_unstemmed | The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line von Xiaoqing Wu |
| title_short | The role of the tumor suppressor Smad4 in the TGF-b [TGF beta] pathway |
| title_sort | the role of the tumor suppressor smad4 in the tgf b tgf beta pathway microarray survey of smad4 dependent target genes in a breast adenocarcinoma cell line |
| title_sub | microarray survey of Smad4 dependent target genes in a breast adenocarcinoma cell line |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=013100635&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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