Differential gene expression in colorectal cancer:
Saved in:
| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2002
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | 78 S. Ill., graph. Darst. : 21 cm |
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MARC
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| 008 | 020924s2002 gw ad|| m||| 00||| eng d | ||
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| 100 | 1 | |a Gelos, Marcos J. |d 1975- |e Verfasser |0 (DE-588)123893844 |4 aut | |
| 245 | 1 | 0 | |a Differential gene expression in colorectal cancer |c vorgelegt von Marcos J. Gelos |
| 264 | 1 | |c 2002 | |
| 300 | |a 78 S. |b Ill., graph. Darst. : 21 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Berlin, Freie Univ., Diss., 2002 | ||
| 650 | 7 | |a Hochschulschrift |2 gtt | |
| 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=009999677&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-009999677 | |
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| _version_ | 1807319675076345856 |
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CONTENTS
1
CONTENTS
1.
INTRODUCTION
5
1.1
DIFFERENT
PATHWAYS
IN
THE
DEVELOPMENT
OF
COLORECTAL
CANCER
5
1.2
MUCINOUS
PHENOTYPE
OF
COLORECTAL
CANCER
7
1.3
ALTERED
GENE
EXPRESSION
IN
COLORECTAL
CANCER
AS
A
MARKER
OF
DIFFERENT
7
PATHWAYS
OF
CARCINOGENESIS
1.4
GENES
WHOSE
EXPRESSION
IS
ALTERED
IN
COLON
CANCER
8
1.5
METHODS
FOR
DETECTION
OF
DIFFERENTIAL
GENE
EXPRESSION
9
2.
OBJECTIVES
OF
THE
WORK
11
3.
MATERIALS
12
3.1.
MOLECULAR
BIOLOGY
12
3.1.1
TISSUE
SAMPLES
12
3.1.2
CELL
LINES
12
3.1.3
CHEMICALS
12
3.1.4
RADIOACTIVE
NUCLEOTIDES
13
3.1.5
KITS
13
3.1.6
ENZYMES
13
3.1.7
PRIMERS
14
3.1.8
PLASMIDS
15
3.1.9
INSTRUMENTS
15
3.2
IMMUNOHISTOCHEMISTRY
16
3.2.1
TISSUE
SAMPLES
16
3.2.2
ANTIBODIES
16
3.2.3
CHEMICALS
16
4.
METHODS
17
4.1
MOLECULAR
BIOLOGY
17
4.1.1
AGAROSE
GEL
ELECTROPHORESIS
17
4.1.2
TOTAL
RNA
ISOLATION
17
4.1.2.1
FROM
PATIENTS
TISSUES
17
4.1.2.2
FROM
CELL
LINES
18
4.1.3
POLY
A
+
RNA
ISOLATION
18
4.1.4
DIFFERENTIAL
HYBRIDIZATION
(ATLAS
ARRAY)
19
A)
RADIOACTIVE
CDNA
SYNTHESIS
19
B)
COLUMN
CHROMATOGRAPHY
19
CONTENTS
2
C)
HYBRIDIZATION
20
D)
WASHING
OF
HYBRIDIZED
MEMBRANES
20
E)
EVALUATION
OF
GENE
EXPRESSION
20
F)
STRIPPING
OF
MEMBRANES
21
4.1.5
RT-PCR
21
4.1.5.1
DNASE
DIGESTION
21
4.1.5.2
CDNA
SYNTHESIS
21
4.1.5.3
PCR
22
4.1.5.4
PCR
PROGRAMMES
23
4.1.6
DNA
ISOLATION
FROM
AGAROSE
GELS
24
4.1.7
PLASMID
ISOLATION
24
4.1.8
NORTHERN
BLOT
24
A)
ELECTROPHORESIS
24
B)
BLOTTING
PROCEDURE
25
4.1.9
RADIOACTIVE
LABELING
OF
DNA
FRAGMENTS
WITH
[A
32
P]
DCTP
26
4.1.10
PURIFICATION
OF
THE
LABELED
PROBE
26
4.1.11
HYBRIDIZATION
26
4.1.11.1
PREHYBRIDIZATION
27
4.1.11.2
HYBRIDIZATION
27
4.1.11.3
WASHING
OF
THE
MEMBRANES
27
4.1.11.4
EXPOSURE
27
4.1.11.5
EVALUATION
OF
THE
FILMS
27
4.1.11.6
STRIPPING
OF
A
MEMBRANE
27
4.2
IMMUNOHISTOCHEMISTRY
28
5.
RESULTS
29
5.1
DETECTION
OF
DIFFERENTIAL
GENE
EXPRESSION
IN
MUCINOUS
AND
NONMUCINOUS
COLORECTAL
CARCINOMA
29
5.1.1
DIFFERENTIAL
GENE
EXPRESSION
PATTERN
IN
MUCINOUS
AND
NONMUCINOUS
COLORECTAL
CARCINOMA
CELL
LINES
DETECTED
BY
CDNA
ARRAY
HYBRIDIZATION
29
5.1.2
VERIFICATION
OF
ATLAS
ARRAY
RESULTS
BY
NORTHERN
BLOT
OR
RT-PCR
33
5.1.2.1
TOPOISOMERASE
II
A
34
5.1.2.2
P21
34
5.1.2.3
WEEIHU
35
5.1.2.4
TFIIH
(44
KDA
SUBUNIT)
35
5.1.2.5
TFIIH
(62
KDA
SUBUNIT)
35
CONTENTS
3
5.1.2.6
CCAAT
DISPLACEMENT
PROTEIN
(CUTL1)
35
5.1.3
EXAMINATION
OF
TISSUE
SAMPLES
FOR
THE
EXPRESSION
OF
GENES
WHOSE
EXPRESSION
IS
ALTERED
IN
CELL
LINES
36
5.1.3.1
TOPOISOMERASE
II
A
37
5.1.3.2
P21
37
5.1.3.3
WEEIHU
5.1.4
EXAMINATION
OF
P21
PROTEIN
EXPRESSION
IN
TISSUES
39
5.2
EXAMINATION
OF
ADDITIONAL
GENES
WHOSE
EXPRESSION
COULD
BE
ALTERED
IN
MUCINOUS
COLON
CARCINOMA
43
5.2.1
MRNA
EXPRESSION
OF
THE
TRANSFORMING
GROWTH
FACTOR-B
TYPE
II
RECEPTOR
40
5.2.1.1
EXPRESSION
IN
CELL
LINES
5.2.1.2
EXPRESSION
IN
TISSUES
5.2.2
MUC6
43
5.2.2.1
EXPRESSION
IN
CELL
LINES
43
5.2.2.2
EXPRESSION
IN
TISSUES
43
5.2.3
FMR-1
44
5.3
SUMMARY
AND
CONCLUSIONS
OF
THE
FIRST
PART
OF
THE
WORK
45
5.4.
TARGET
GENES
OF
THE
B-CATENIN-TCITLEF
SIGNALING
PATHWAY
IN
COLORECTAL
CARCINOMA
46
5.4.1
CHARACTERIZATION
OF
THE
MODEL
CELL
LINES
46
5.4.2
TARGET
GENES
OF
THE
B-CATENIN-TCF/LEF
PATHWAY
IN
COLORECTAL
CARCINOMA
CELLS
5.4.2.1
ALTERED
GENE
EXPRESSION
AFTER
B-CATENIN
TRANSFECTION
DETECTED
BY
ATLAS
48
ARRAY
HYBRIDIZATION
5.4.2.2
VERIFICATION
OF
THE
OBTAINED
RESULTS
BY
NORTHERN
BLOT
HYBRIDIZATION
49
5.4.2.3
ZO-1
AND
UPAR
PROTEIN
EXPRESSION
IN
B-CATENIN-TRANSFECTED
AND
51
CONTROL
CELLS
5.4.2.4
ZO-1
EXPRESSION
IN
CELL
LINES
WITH
HIGH
ENDOGENOUS
B-CATENIN
LEVELS
51
5.4.2.5
EXAMINATION
OF
UPAR
AND
B-CATENIN
EXPRESSION
IN
PRIMARY
TUMOURS
AND
METASTASES
OF
PATIENTS
WITH
COLORECTAL
CARCINOMAS
AT
UICC
STAGE
IV
52
5.5
SUMMARY
AND
CONCLUSION
OF
THE
SECOND
PART
OF
THE
WORK
54
6.
DISCUSSION
52
6.1
CDNA
ARRAY
HYBRIDIZATION
AS
METHOD
FOR
DETECTION
OF
ALTERED
GENE
EXPRESSION
52
6.2.1
ALTERATIONS
IN
GENE
EXPRESSION
CHARACTERISTIC
FOR
MUCINOUS
COLON
53
CONTENTS
4
CARCINOMA
CELLS
6.2.1.1
P21
OVEREXPRESSION
IN
MUCINOUS
CARCINOMAS
OF
THE
COLORECTUM
-
DOES
IT
INHIBIT
TUMOUR
GROWTH?
53
6.2.1.2
DOES
A
RELATIONSHIP
BETWEEN
P2
1
AND
TGFB-RII
EXPRESSION
EXIST
IN
THE
MUCINOUS
PHENOTYPE
OF
COLON
CANCER?
54
6.2.1.3
EXPRESSION
OF
MUC6
GENE
55
6.3
ALTERATIONS
IN
GENE
EXPRESSION
COMMON
TO
MUCINOUS
AND
NONMUCINOUS
COLON
CANCER
CELLS
56
6.3.1
POTENTIAL
CONSEQUENCES
OF
THE
LOSS
OF
THE
CHECKPOINT-CONTROLLING
GENE
WEEIHU
56
6.3.2
OTHER
TUMOUR-ASSOCIATED
ALTERATIONS
OF
GENE
EXPRESSION
DETECTED
IN
THE
PRESENT
WORK
57
6.4
FUTURE
PERSPECTIVES
58
6.5.
IDENTIFICATION
OF
TARGET
GENES
OF
THE
B-CATENIN-TCF/LEF
SIGNALING
PATHWAY
6.5.1
ROLE
OF
B-CATENIN
IN
COLORECTAL
CARCINOGENESIS
59
6.5.2
SIGNIFICANCE
OF
THE
IDENTIFIED
TARGET
GENES
FOR
COLONIC
TUMOURIGENESIS
59
6.5.2.1
BIOLOGICAL
CONSEQUENCES
OF
C-JUN,
FRA-1
AND
UPAR
EXPRESSION
59
6.5.2.2
SUPPRESSION
OF
ZO-1
60
6.5.2.3
FUTURE
PERSPECTIVES
61
7.
SUMMARY
62
8.
REFERENCES
63
9.
PUBLICATIONS
76
10.
ACKNOWLEDGEMENTS
77 |
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| author_GND | (DE-588)123893844 |
| author_facet | Gelos, Marcos J. 1975- |
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| indexdate | 2024-08-14T00:12:34Z |
| institution | BVB |
| language | English |
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| physical | 78 S. Ill., graph. Darst. : 21 cm |
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| spelling | Gelos, Marcos J. 1975- Verfasser (DE-588)123893844 aut Differential gene expression in colorectal cancer vorgelegt von Marcos J. Gelos 2002 78 S. Ill., graph. Darst. : 21 cm txt rdacontent n rdamedia nc rdacarrier Berlin, Freie Univ., Diss., 2002 Hochschulschrift gtt (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=009999677&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Gelos, Marcos J. 1975- Differential gene expression in colorectal cancer Hochschulschrift gtt |
| subject_GND | (DE-588)4113937-9 |
| title | Differential gene expression in colorectal cancer |
| title_auth | Differential gene expression in colorectal cancer |
| title_exact_search | Differential gene expression in colorectal cancer |
| title_full | Differential gene expression in colorectal cancer vorgelegt von Marcos J. Gelos |
| title_fullStr | Differential gene expression in colorectal cancer vorgelegt von Marcos J. Gelos |
| title_full_unstemmed | Differential gene expression in colorectal cancer vorgelegt von Marcos J. Gelos |
| title_short | Differential gene expression in colorectal cancer |
| title_sort | differential gene expression in colorectal cancer |
| topic | Hochschulschrift gtt |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009999677&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT gelosmarcosj differentialgeneexpressionincolorectalcancer |