Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy:
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| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
2000
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | 163 S. Ill., graph. Darst. |
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MARC
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| 008 | 010418s2000 ad|| m||| 00||| eng d | ||
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| 035 | |a (OCoLC)48739969 | ||
| 035 | |a (DE-599)BVBBV013685793 | ||
| 040 | |a DE-604 |b ger |e rakwb | ||
| 041 | 0 | |a eng | |
| 049 | |a DE-91 |a DE-355 |a DE-188 | ||
| 100 | 1 | |a Leech, Siân |d 1971- |e Verfasser |0 (DE-588)122808789 |4 aut | |
| 245 | 1 | 0 | |a Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy |c vorgelegt von Siân Leech |
| 264 | 1 | |c 2000 | |
| 300 | |a 163 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |a Stuttgart, Univ., Diss., 2000 | ||
| 650 | 7 | |a Hochschulschrift |2 gtt | |
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| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-009352230 | |
Record in the Search Index
| _version_ | 1807504454122995712 |
|---|---|
| adam_text |
1.0
C
ONTENTS
2
1.0
C
ONTENTS
2.0
A
BBREVIATIONS
.
7
3.0
S
UMMARY
.
8
4.0
Z
USAMMENFASSUNG
.
10
5.0
I
NTRODUCTION
.
19
5.1
LUNG
CANCER
.
19
5.2
CURRENT
CONVENTIONAL
THERAPIES
FOR
LUNG
CANCER
TREATMENT
.
20
5.2.1
NON-SMALL
CELL
LUNG
CANCER
.
20
5.2.2
SMALL
CELL
LUNG
CANCER
.
20
53
APOPTOSIS
.
21
5.3.1
THE
BCL-2
PROTEIN
FAMILY
.
23
5.3.2
THE
CASPASE
FAMILY
.
28
5.3.3
APOPTOTIC
PATHWAYS:
DEATH
RECEPTOR,
OR
MITOCHONDRIA
MEDIATED
.
30
5.3.3.1
MITOCHONDRIAL
MEDIATED
APOPTOSIS
.
31
5.3.3.2
RECEPTOR
MEDIATED
APOPTOSIS
.
33
5.4
ANTISENSE
OLIGONUCLEOTIDES:
REASONS,
MECHANISMS
AND
ACHIEVEMENTS
.
34
5.4.1
MECHANISM
OF
ACTION
OF
ANTISENSE
OLIGONUCLEOTIDES
.
35
5.4.2
STABILITY
OF
ANTISENSE
OLIGONUCLEOTIDES
(PHOSPHOROTHIOATE
OLIGONUCLEOTIDE
DEVELOPMENT)
.
37
5.4.3
DEVELOPMENT
OF
"
SECOND
GENERATION
" ANTISENSE
OLIGONUCLEOTIDES
.
39
5.4.4
ANTISENSE
OLIGONUCLEOTIDE
DELIVERY
.
40
5.5
EXPERIMENTAL
AIMS
AND
OBJECTIVES
.
41
6.0
M
ATERIALS
AND
M
ETHODS
.
43
6.1
CELL
LINES
.
43
6.2
ANTISENSE
OLIGONUCLEOTIDES
.
43
6.3
ANTISENSE
OLIGONUCLEOTIDE
TREATMENT
OF
CELLS
.
45
6.4
WESTERN
BLOT
ANALYSIS
.
47
6.4.1
COLLECTION
OF
CELLS/SAMPLE
PREPARATION
.
47
6.4.2
DETERMINATION
OF
PROTEIN
CONCENTRATION
WITHIN
SAMPLES
.
48
6.4.3
PREPARATION
AND
RUNNING
OF
SDS-PAGE
GEL
.
48
6.4.4
BLOTTING
OF
PROTEINS
ONTO
PVF-MEMBRANE
.
50
1.0
C
ONTENTS
3
6.4.5
BLOCKING
AND
PROBING
OF
MEMBRANES
.
50
6.4.6
ANALYSIS
OF
MEMBRANES
.
50
6.5
REAL
TIME
REVERSE
TRANSCRIPTION
POLYMERASE
CHAIN
REACTION
.
52
6.5.1
CELL
PELLET
COLLECTION
AND
RNA
ISOLATION
.
53
6.5.2
PREPARATION
OF
CDNA
.
54
6.5.3
REAL
TIME
PCR
ANALYSIS
.
55
6.6
DETERMINATION
OF
CELL
VIABILITY:
MTT
PROLIFERATION
ASSAY
.
58
6.6.1
CELL
VIABILITY
FOLLOWING
OLIGONUCLEOTIDE
TREATMENT
.
58
6.6.2
CELL
VIABILITY
FOLLOWING
OLIGONUCLEOTIDE
TREATMENT
IN
COMBINATION
WITH
CHEMOTHERAPY
.
59
6.7
IMMUNOCYTOCHEMISTRY
.
60
6.8
APOPTOSIS
ASSAYS
.
61
6.8.1
NUCLEAR
MORPHOLOGY
ASSAYS
.
61
6.8.2
DETECTION
OF
MITOCHONDRIAL
TRANS-MEMBRANE
POTENTIAL
DISRUPTION
.
62
6.8.3
DETECTION
OF
CASPASE
3-LIKE
PROTEASE
ACTIVITY
.
63
7.0
R
ESULTS
.
65
7.1
OPTIMISATION
OF
IN
VITRO
TRANSFECTION
PROTOCOLS
.
65
7.2
TREATMENT
OF
A549
NSCLC
CELLS
WITH
FIRST
GENERATION
PHOSPHOROTHIOATE
OLIGONUCLEOTIDE
3011
TARGETING
BCL-XL
.
68
7.2.1
DETERMINATION
OF
CELL
PROLIFERATION
FOLLOWING
TREATMENT
OF
A549
CELLS
WITH
PHOSPHOROTHIOATE
ANTISENSE
OLIGONUCLEOTIDE
3011
.
68
7.2.2
EFFECT
OF
TREATMENT
OF
A549
NSCLC
CELLS
WITH
PHOSPHOROTHIOATE
ANTISENSE
OLIGONUCLEOTIDE
3011
ON
BCL-XL
PROTEIN
LEVELS
.
69
73
TREATMENT
OF
A549
NSCLC
CELLS
WITH
SECOND
GENERATION
2
'
-MOE
PHOSPHOROTHIOATE
OLIGONUCLEOTIDE
4259
.
70
7.3.1
EFFECT
ON
PROLIFERATION
OF
A549
NSCLC
CELLS
FOLLOWING
TREATMENT
WITH
ANTISENSE
OLIGONUCLEOTIDE
4259
.
70
7.3.2
LEVELS
OF
BCL-XL
IN
A549
CELLS
FOLLOWING
TREATMENT
WITH
OLIGONUCLEOTIDE
4259
.
'
.
72
7.3.3
EFFECT
OF
TREATMENT
WITH
OLIGONUCLEOTIDE
4259
ON
MORPHOLOGY
.
OF
A549
NSCLC
CELLS
.
73
7.3.4
NUCLEAR
MORPHOLOGY
OF
A549
NSCLC
CELLS
FOLLOWING
TREATMENT
WITH
OLIGONUCLEOTIDE
4259
.
74
7.3.5
ACTIVATION
OF
CASPASE
3-LIKE
PROTEASES
IN
A549
CELLS
FOLLOWING
TREATMENT
1.0
C
ONTENTS
4
WITH
OLIGONUCLEOTIDE
4259
.
75
7.4
TREATMENT
OF
ADDITIONAL
LUNG
CANCER
CELL
LINES
WITH
OLIGONUCLEOTIDE
4259
TARGETING
BCL-XL
.
76
7.4.1
BASAL
LEVELS
OF
BCL-2
AND
BCL-XL
ANTI-APOPTOTIC
PROTEINS
IN
LUNG
CANCER
CELL
LINES
TESTED
.
76
7.4.2
EFFECT
OF
OLIGONUCLEOTIDE
4259
TREATMENT
ON
PROLIFERATION
OF
ADDITIONAL
LUNG
CANCER
CELL
LINES
.
77
7.4.3
EFFECT
OF
OLIGONUCLEOTIDE
4259
TREATMENT
ON
BCL-2
AND
BCL-XL
PROTEIN
LEVELS
WITHIN
ADDITIONAL
LUNG
CANCER
CELL
LINES
.
80
7.4.4
ACTIVATION
OF
CASPASE
3-LIKE
PROTEASES
IN
ADDITIONAL
CELL
LINES
TESTED
(NCI-H125,
SW2,
AND
NCI-H69)
FOLLOWING
TREATMENT
WITH
OLIGONUCLEOTIDE
4259
.
82
7.5
DEVELOPMENT
OF
A
FIRST
GENERATION
PHOSPHOROTHIOATE
OLIGONUCLEOTIDE
TARGETING
BOTH
BCL-2
AND
BCL-XL
EXPRESSION
SIMULTANEOUSLY
.
83
7.6
DEVELOPMENT
OF
THE
SECOND
GENERATION
"
BI-SPECIFIC
"
2
'
-MOE
PHOS
PHOROTHIOATE
ANTISENSE
OLIGONUCLEOTIDES
4625
AND
4627
.
86
7.6.1
EFFICACY
OF
MODIFIED
BI-SPECIFIC
OLIGONUCLEOTIDES
ON
CELL
PROLIFERATION
.
86
7.6.2
LEVELS
OF
BCL-2
AND
BCL-XL
PROTEIN
IN
SW2
SCLC
CELLS
FOLLOWING
TREATMENT
WITH
BI-SPECIFIC
ANTISENSE
OLIGONUCLEOTIDES
4625
AND
4627
.
87
7.6.3
INVESTIGATIONS
INTO
THE
EFFECT
OF
TREATMENT
OF
SW2
SCLC
CELLS
WITH
BI-SPECIFIC
OLIGONUCLEOTIDES
4625
AND
4627
ON
RNRNA
LEVELS
.
88
7.6.4
NUCLEAR
MORPHOLOGY
OF
SW2
SCLC
CELLS
FOLLOWING
TREATMENT
WITH
BI-SPECIFIC
OLIGONUCLEOTIDES
4625
AND
4627
.
91
7.6.5
ANALYSIS
OF
ACTIVATION
OF
CASPASE
3-LIKE
PROTEASES
FOLLOWING
TREATMENT
WITH
BI-SPECIFIC
ANTISENSE
OLIGONUCLEOTIDES
4625
AND
4627
.
92
7.7
TREATMENT
OF
ADDITIONAL
LUNG
CANCER
CELL
LINES
(A549
AND
NCI-H125)
WITH
BI-SPECIFIC
OLIGONUCLEOTIDE
4625
AND
MONO-SPECIFIC
OLIGONUCLEOTIDE
4259
.
92
7.7.1
MTT
PROLIFERATION
ASSAYS
OF
SW2,
NCLH125
AND
A549
LUNG
CANCER
CELL
LINES
FOLLOWING
OLIGONUCLEOTIDE
TREATMENT
.
93
7.7.2
DOWN-REGULATION
OF
BCL-XL
AND
BCL-2
PROTEIN
IN
A549,
NCI-H125
AND
SW2
CELL
LINES
FOLLOWING
TREATMENT
WITH
OLIGONUCLEOTIDES
4259
AND
4625
.
94
7.7.3
IMMUNOCYTOCHEMICAL INVESTIGATIONS
INTO
THE
LEVELS
OF
BCL-XL
PROTEIN
REMAINING
IN
A549
AND
NCI-H125
CELLS
FOLLOWING
TREATMENT
WITH
OLIGONUCLEOTIDE
4625
.
96
1.0
C
ONTENTS
5
7.7.4
CELLULAR
MORPHOLOGY
FOLLOWING
TREATMENT
WITH
ANTISENSE
OLIGONUCLEOTIDES
.
98
7.7.5
NUCLEAR
MORPHOLOGY
OF
A549
AND
NCI-H125
LUNG
CANCER
CELLS
FOLLOWING
TREATMENT
WITH
BI-SPECIFIC
ANTISENSE
OLIGONUCLEOTIDE
4625
.
99
7.7.6
CASPASE
3-LIKE
PROTEASE
ACTIVITY
IN
LUNG
CANCER
CELL
LINES
FOLLOWING
TREATMENT
WITH
OLIGONUCLEOTIDES
4625
AND
4259
.
100
7.7.7
ANALYSIS
OF
LOSS
OF
MITOCHONDRIAL
TRANS-MEMBRANE
POTENTIAL
IN
NSCLC
CELL
LINES
FOLLOWING
TREATMENT
WITH
BI-SPECIFIC OLIGONUCLEOTIDE
4625
.
101
7.8
THE
TREATMENT
OF
LUNG
CANCER
CELL
LINES
WITH
OLIGONUCLEOTIDES
4259
AND
4625
IN
COMBINATION
WITH
CONVENTIONAL
CHEMOTHERAPY
.
104
7.8.1
COMBINATION
OF
OLIGONUCLEOTIDES
AND
CHEMOTHERAPEUTIC
DRUGS
ON
A549
NSCLC
CELLS
.
105
7.8.1.1
COMBINATION
TREATMENT
OF
A549
NSCLC
CELLS
WITH
TAXOL
AND
OLIGONUCLEOTIDES
4259
OR
4625
.
105
7.8.1.2
COMBINATION
TREATMENT
OF
A549
NSCLC
CELLS
WITH
CISPLATIN
AND
OLIGONUCLEOTIDES
4259
OR
4625
.
107
7.8.2
COMBINATION
OF
OLIGONUCLEOTIDES
AND
CHEMOTHERAPEUTIC
DRUGS
ON
NC1-H125
NSCLC
CELLS
AND
SW2
SCLC
CELLS
.
109
7.8.2.1
COMBINATION
TREATMENT
OF
LUNG
CANCER
CELL
LINES
WITH
OLIGONUCLEOTIDES
AND
TAXOL
.
109
7.8.2.2
COMBINATION
TREATMENT
OF
LUNG
CANCER
CELL
LINES
WITH
OLIGONUCLEOTIDES
AND
CISPLATIN
.
110
7.9
TREATMENT
OF
LUNG
AND
BREAST
CANCER
CELL
LINES
WITH
OLIGONUCLEOTIDES
4259
AND
4625
IN
COMBINATION
WITH
RADIOTHERAPY
.
ILL
7.9.1
THE
EFFECT
OF
RADIATION/OLIGONUCLEOTIDE
COMBINATION
TREATMENT
ON
CELL
PROLIFERATION
.
112
7.9.2
EFFECT
OF
RADIATION/OLIGONUCLEOTIDE
COMBINATION
TREATMENT
ON
PROTEIN
EXPRESSION
.
116
7.9.2.1
TREATMENT
OF
SW2
SCLC
CELLS
.
116
7.9.2.2
TREATMENT
OF
MDA-MB-23
1
BREAST
CANCER
CELLS
.
118
7.9.2.3
TREATMENT
OFNSCLC
CELL
LINES
A549
AND
NCI-H125
.
120
7.9.3
EFFECT
OF
RADIATION/OLIGONUCLEOTIDE
COMBINATION
TREATMENT
ON
NUCLEAR
MORPHOLOGY
OF
LUNG
CANCER
CELL
LINES
.
121
8.0
D
ISCUSSION
.
124
8.1
TREATMENT
OF
A549
NSCLC
CELLS
WITH
FIRST
GENERATION
PHOSPHOROTHIOATE
OLIGONUCLEOTIDE
3011
TARGETING
BCL-XL
.
124
8.2
DEVELOPMENT
AND
TESTING
OF
SECOND
GENERATION
2
'
-MOE
PHOSPHOROTHIOATE
OLIGONUCLEOTIDES
4259
AND
SC4259
.
126
1.0
C
ONTENTS
6
83
INVESTIGATING
THE
THERAPEUTIC
POTENTIAL
OF
OLIGONUCLEOTIDE
4259
.
128
8.4
DEVELOPMENT
OF
A
"
BI-SPECIFIC
"
OLIGONUCLEOTIDE
TARGETING
BCL-2
AND
BCL-XL
NIRNA
EXPRESSION
SIMULTANEOUSLY
.
131
8.5
INVESTIGATING
THE
THERAPEUTIC
POTENTIAL
OF
OLIGONUCLEOTIDE
4625
.
133
8.6
INVESTIGATIONS
INTO
THE
USE
OF
OLIGONUCLEOTIDE
4259,
OR
OLIGONUCLEOTIDE
4625
IN
COMBINATION
WITH
CHEMOTHERAPEUTIC
AGENTS
.
136
8.7
COMBINATION
OF
OLIGONUCLEOTIDES
4259
AND
4625
WITH
RADIATION
TREATMENT:
A
NEW
DIRECTION
IN
ANTISENSE
OLIGONUCLEOTIDE
TREATMENT
.
139
8.8
MECHANISM
OF
ACTION
OF
ANTISENSE
OLIGONUCLEOTIDES
?
.
143
8.9
CONCLUSIONS
.
144
9.0
R
EFERENCES
.
145
10.0
C
URRICULUM
V
ITAE
.
162
11.0
A
CKNOWLEDGEMENTS
.
163 |
| any_adam_object | 1 |
| author | Leech, Siân 1971- |
| author_GND | (DE-588)122808789 |
| author_facet | Leech, Siân 1971- |
| author_role | aut |
| author_sort | Leech, Siân 1971- |
| author_variant | s l sl |
| building | Verbundindex |
| bvnumber | BV013685793 |
| ctrlnum | (OCoLC)48739969 (DE-599)BVBBV013685793 |
| format | Thesis Book |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV013685793 |
| illustrated | Illustrated |
| indexdate | 2024-08-16T01:09:33Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-009352230 |
| oclc_num | 48739969 |
| open_access_boolean | |
| owner | DE-91 DE-BY-TUM DE-355 DE-BY-UBR DE-188 |
| owner_facet | DE-91 DE-BY-TUM DE-355 DE-BY-UBR DE-188 |
| physical | 163 S. Ill., graph. Darst. |
| publishDate | 2000 |
| publishDateSearch | 2000 |
| publishDateSort | 2000 |
| record_format | marc |
| spelling | Leech, Siân 1971- Verfasser (DE-588)122808789 aut Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy vorgelegt von Siân Leech 2000 163 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Stuttgart, Univ., Diss., 2000 Hochschulschrift gtt Apoptosis (DE-588)4291096-1 gnd rswk-swf Krebszelle (DE-588)4128704-6 gnd rswk-swf Lungenkrebs (DE-588)4036668-6 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Lungenkrebs (DE-588)4036668-6 s Krebszelle (DE-588)4128704-6 s Apoptosis (DE-588)4291096-1 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009352230&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Leech, Siân 1971- Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy Hochschulschrift gtt Apoptosis (DE-588)4291096-1 gnd Krebszelle (DE-588)4128704-6 gnd Lungenkrebs (DE-588)4036668-6 gnd |
| subject_GND | (DE-588)4291096-1 (DE-588)4128704-6 (DE-588)4036668-6 (DE-588)4113937-9 |
| title | Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy |
| title_auth | Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy |
| title_exact_search | Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy |
| title_full | Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy vorgelegt von Siân Leech |
| title_fullStr | Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy vorgelegt von Siân Leech |
| title_full_unstemmed | Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy vorgelegt von Siân Leech |
| title_short | Treatment of lung cancer cells with bcl-2 and/or bcl-xL antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy |
| title_sort | treatment of lung cancer cells with bcl 2 and or bcl xl antisense oligonucleotides induces apoptosis and sensitises cells to conventional therapy |
| topic | Hochschulschrift gtt Apoptosis (DE-588)4291096-1 gnd Krebszelle (DE-588)4128704-6 gnd Lungenkrebs (DE-588)4036668-6 gnd |
| topic_facet | Hochschulschrift Apoptosis Krebszelle Lungenkrebs |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009352230&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT leechsian treatmentoflungcancercellswithbcl2andorbclxlantisenseoligonucleotidesinducesapoptosisandsensitisescellstoconventionaltherapy |