LBH589-induced DAPK activation triggers apoptosis and autophagy in human colon tumor cells: = Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
|
| Schlagworte: | |
| Online-Zugang: | kostenfrei http://d-nb.info/1033029874/34 kostenfrei Inhaltsverzeichnis |
| Beschreibung: | XIV, 90 S. Ill., graph. Darst. |
Internformat
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| 264 | 1 | |c 2013 | |
| 300 | |a XIV, 90 S. |b Ill., graph. Darst. | ||
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Datensatz im Suchindex
| _version_ | 1804150315233050624 |
|---|---|
| adam_text | IMAGE 1
INDEX
ABBREVIATIONS VIII
ABSTRACT XI
ZUSAMMENFASSUNG XIII
1. INTRODUCTION 1
1.1 CHROMATIN STRUCTURE 1
1.2 EPIGENETICS AND CANCER 2
1.3 HISTONE DEACETYLASES (HDAC) 2
1.4 HISTONE ACETYL TRANSFERASES (HAT) 3
1.5 DNA METHYLATION 3
1.6 DNA METHYL TRANSFERASES (DNMTS) 4
1.7 HISTONE DEACETYLASE INHIBITORS (HDACI) AS ANTI-CANCER AGENTS 4
1.7.1 HISTONE DEACETYLASE INHIBITORS (HDACI) 4
1.7.2 LBH589 (PANOBINOSTAT) 6
1.8 DEATH ASSOCIATED PROTEIN KINASE (DAPK) 7
1.8.1 DAPK STRUCTURE 7
1.8.2 DAPK FUNCTION 8
1.9 PROGRAMMED CELL DEATH PATHWAYS 9
1.9.1 APOPTOSIS 9
1.9.2 AUTOPHAGY 10
2. AIMS O F THE WORK 14
3. MATERIALS AND METHODS 16
3.1 MATERIALS 16
3.1.1 CHEMICALS AND REAGENTS 16
3.1.2 CELL CULTURE 18
3.1.3 CELL LINES 18
3.1.4 BUFFERS AND SOLUTIONS 18
3.1.5 ANTIBODIES 20
3.1.6 DRUGS AND CHEMICALS 21
3.1.7 INHIBITORS 21
3.1.8 KITS 21
3.1.9 EQUIPMENTS 22
3.1.10 CONSUMABLES 23
IV
HTTP://D-NB.INFO/1035263769
IMAGE 2
INDEX
3.1.11 SOFTWARE 2 3
3.1.12 DATABASE 23
3.2 METHODS 24
3.2.1 TUMOR CELL LINES AND CELL CULTURE 24
3.2.2 GENERATION O F DAPK SHRNA STABLE CELL LINE 24
3.2.3 GENERATION O F DAPK+++ OVEREXPRESSING CELL LINE 24
3.2.4 XCELLIGENCE REAL-TIME CELL ASSAY 25
3.2.5 MTT ASSAY 25
3.2.6 CRYSTAL VIOLET ASSAY 25
3.2.7 CLONOGENIC ASSAY 26
3.2.8 WESTERN BLOTTING 26
3.2.8.1 DETERMINATION O F PROTEIN CONCENTRATION 26
3.2.8.2 WESTERN BLOT ANALYSIS 27
3.2.9 DETECTION O F AUTOPHAGIC VACUOLES BY ACRIDINE ORANGE 28
3.2.10 DETECTION O F AUTOPHAGOSOMES BY LC3 IMMUNOFLUORESCENCE STAINING
28
3.2.11 FLOW CYTOMETRIC DETECTION O F APOPTOSIS BY ANNEXIN V-FITC
LABELLING 28
3.2.12 FLOW CYTOMETRIC ANALYSIS O F CELL CYCLE DISTRIBUTION 28
3.2.13 IMMUNOPRECIPITATION 29
3.2.14 IN VITRO KINASE ASSAY 29
3.2.15 RNA ISOLATION AND CDNA SYNTHESIS 29
3.2.16 REAL-TIME REVERSE TRANSCRIPTION-PCR 30
3.2.17 IN VIVO EXPERIMENTAL METHODS 30
3.2.17.1 XENOGRAFT MODEL 30
3.2.17.2 TISSUE MICROARRAY AND IMMUNOHISTOCHEMISTRY 31
3.2.17.3 ASSESSMENT O F IMMUNOHISTOCHEMICAL PROTEIN EXPRESSION 31
3.2.17.4 PREPARATION O F ANIMALS FOR MICROPET IMAGING 31
3.2.17.5 MICROPET IMAGING 32
3.2.18 STATISTICAL ANALYSIS 32
4. RESULTS 33
4.1 LBH589 DECREASES CELL VIABILITY IN DAPK WT CELLS 33
4.2 GENERATION O F DAPK KNOCKDOWN STABLE CELL LINE (DAPK SHRNA) IN
DAPK WT CELLS 34
V
IMAGE 3
INDEX
4.3 GENERATION O F TAMOXIFEN-INDUCIBLE DAPK OVEREXPRESSION STABLE CELL
LINE
(DAPK+++) IN DAPK WT CELLS 34
4.4 EFFECT O F TAMOXIFEN AND/OR LBHS89 ON THE EXPRESSION O F DAPK
IN DAPK+++CELLS 35
4.5 LBH589 INDUCES ACETYLATION O F HISTONES H3 AND H4 IN HUMAN COLON
TUMOR CELLS.. 36
4.6 KNOCKDOWN AND OVEREXPRESSION O F DAPK IN DAPK WT CELLS DID NOT
INFLUENCE
CELL PROLIFERATION 37
4.7 LBHS89 INHIBITS CELL PROLIFERATION IN HUMAN COLON TUMOR CELLS HAVING
DIFFERENT
DAPK LEVELS 38
4.8 LBH589 REDUCES THE LONG-TERM SURVIVAL IN DAPK WT CELLS HAVING
DIFFERENT
DAPK LEVELS 39
4.9 LBH589 INDUCES UP-REGULATION AND ACTIVATION O F DAPK IN HUMAN COLON
CANCER
CELLS IN VITRO 40
4.10 ROLE O F DAPK IN LBH589-INDUCED APOPTOSIS 42
4.10.1 LBHS89 INDUCES CELL CYCLE ARREST IN HUMAN COLON TUMOR CELLS 42
4.10.2 LBH589 CAUSES FORMATION O F APOPTOTIC BODIES IN A
DAPK-INDEPENDENT
MANNER 43
4.10.3 LBH589 ACTIVATES CASPASES AND INDUCES APOPTOSIS IN A
DAPK-INDEPENDENT
MANNER 4 3
4.10.4 ZVAD RESCUED THE CELLS FROM LBH589 INDUCED APOPTOSIS IN HUMAN
COLON
TUMOR CELLS 46
4.10.5 LBH589 ALSO INDUCES CASPASE INDEPENDENT CELL DEATH IN HUMAN COLON
TUMOR CELLS 46
4.10.6 LBH589-INDUCED APOPTOSIS IS INDEPENDENT O F DAPK KINASE ACTIVITY
4 7
4.11 ROLE O F DAPK IN LBH589-INDUCED AUTOPHAGY 49
4.11.1 LBH589 INDUCES AUTOPHAGY IN HUMAN COLON CANCER CELLS IN A DAPK
-DEPENDENT MANNER 49
4.11.2 EFFECT O F LBH589 ON OTHER AUTOPHAGY MARKERS 51
4.11.3 INHIBITION O F AUTOPHAGY DECREASED ACIDIFICATION O F VESICULAR
ORGANELLES IN
HUMAN COLON TUMOR CELLS 51
4.11.4 INHIBITION O F AUTOPHAGY CAUSES ACCUMULATION O F LC3-II AND P62
PROTEIN.... 52
4.11.5 CROSSTALK BETWEEN LBH589-INDUCED APOPTOSIS AND AUTOPHAGY 53
4.12 DAPK UP-REGULATION OR ACTIVATION IS IMPORTANT FOR THE PRO-APOPTOTIC
EFFECTS OF
LBH589 IN DIFFERENT COLORECTAL CANCER CELLS 55
VI
IMAGE 4
INDEX
4.13 DAPK UP-REGULATION AND ACTIVATION IS A GENERAL PHENOMENON AND IS
CORRELATED
WITH THE MODE O F CELL DEATH AFTER TREATMENT WITH HDACI . . 57
4.14 IN VIVO COLON TUMOR XENOGRAFTS 59
4.14.1 LBH589 SUPPRESSES THE GROWTH O F COLON TUMOR XENOGRAFTS 59
4.14.2 LBH589 UP-REGULATES DAPK IN COLON TUMOR XENOGRAFTS 60
4.14.3 LBH589 REDUCES PROLIFERATION IN DAPK WT AND DAPK SHRNA
XENOGRAFTS.. 60
4.14.4 LBH589 DOES NOT SUPPRESS GROWTH O F HT29 COLON TUMOR XENOGRAFTS
61
4.14.5 UPET IMAGING WITH AN 18F-FLUOROGLYCOSYLATED RGD PEPTIDE
([18F]FGLC-RGD)
IN HT29-XENOGRAFT MICE 62
5. DISCUSSION 64
5.1 LBH589 INDUCES THE ACETYLATION O F CORE HISTONES H3 AND H4 65
5.2 LBH589 UP-REGULATES TUMOR SUPPRESSOR DAPK EXPRESSION IN VITRO AND IN
VIVO.. 66
5.3 LBH589 REDUCES THE PROLIFERATION IN VITRO AND IN VIVO AND CAUSES
G2/M CELL CYCLE
ARREST IN A DAPK-INDEPENDENT MANNER 67
5.4 LBH589 CAUSES DNA DAMAGE 68
5.5 LBH589 INDUCES APOPTOSIS IN A DAPK-INDEPENDENT MANNER 68
5.6 LBH589 INDUCES AUTOPHAGY IN A DAPK-DEPENDENT MANNER 70
5.7 CONCLUSION AND OUTLOOK 73
6. REFERENCES 75
7. CURRICULUM VITAE 88
7.1 LIST O F ORIGINAL RESEARCH PUBLICATIONS 88
7.2 POSTER/ORAL PRESENTATIONS 89
VII
|
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| dewey-ones | 610 - Medicine and health |
| dewey-raw | 610 |
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| dewey-sort | 3610 |
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| discipline | Medizin |
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| spelling | Gandesiri, Muktheshwar Verfasser aut LBH589-induced DAPK activation triggers apoptosis and autophagy in human colon tumor cells = Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen von Muktheshwar Gandesiri Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen 2013 XIV, 90 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Erlangen-Nürnberg, Univ., Diss., 2013 Apoptosis (DE-588)4291096-1 gnd rswk-swf Autophagie Physiologie (DE-588)4441664-7 gnd rswk-swf Dickdarmkrebs (DE-588)4012122-7 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Dickdarmkrebs (DE-588)4012122-7 s Apoptosis (DE-588)4291096-1 s DE-604 Autophagie Physiologie (DE-588)4441664-7 s https://open.fau.de/handle/openfau/3238 Verlag kostenfrei Volltext http://d-nb.info/1033029874/34 Langzeitarchivierung Nationalbibliothek https://nbn-resolving.org/urn:nbn:de:bvb:29-opus-44701 Resolving-System kostenfrei Volltext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025969781&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Gandesiri, Muktheshwar LBH589-induced DAPK activation triggers apoptosis and autophagy in human colon tumor cells = Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen Apoptosis (DE-588)4291096-1 gnd Autophagie Physiologie (DE-588)4441664-7 gnd Dickdarmkrebs (DE-588)4012122-7 gnd |
| subject_GND | (DE-588)4291096-1 (DE-588)4441664-7 (DE-588)4012122-7 (DE-588)4113937-9 |
| title | LBH589-induced DAPK activation triggers apoptosis and autophagy in human colon tumor cells = Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen |
| title_alt | Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen |
| title_auth | LBH589-induced DAPK activation triggers apoptosis and autophagy in human colon tumor cells = Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen |
| title_exact_search | LBH589-induced DAPK activation triggers apoptosis and autophagy in human colon tumor cells = Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen |
| title_full | LBH589-induced DAPK activation triggers apoptosis and autophagy in human colon tumor cells = Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen von Muktheshwar Gandesiri |
| title_fullStr | LBH589-induced DAPK activation triggers apoptosis and autophagy in human colon tumor cells = Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen von Muktheshwar Gandesiri |
| title_full_unstemmed | LBH589-induced DAPK activation triggers apoptosis and autophagy in human colon tumor cells = Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen von Muktheshwar Gandesiri |
| title_short | LBH589-induced DAPK activation triggers apoptosis and autophagy in human colon tumor cells |
| title_sort | lbh589 induced dapk activation triggers apoptosis and autophagy in human colon tumor cells die lbh589 induzierte dapk aktivierung fordert apoptose und autophagie in humanen dickdarmkrebszellen |
| title_sub | = Die LBH589-induzierte DAPK-Aktivierung fördert Apoptose und Autophagie in humanen Dickdarmkrebszellen |
| topic | Apoptosis (DE-588)4291096-1 gnd Autophagie Physiologie (DE-588)4441664-7 gnd Dickdarmkrebs (DE-588)4012122-7 gnd |
| topic_facet | Apoptosis Autophagie Physiologie Dickdarmkrebs Hochschulschrift |
| url | https://open.fau.de/handle/openfau/3238 http://d-nb.info/1033029874/34 https://nbn-resolving.org/urn:nbn:de:bvb:29-opus-44701 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=025969781&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT gandesirimuktheshwar lbh589induceddapkactivationtriggersapoptosisandautophagyinhumancolontumorcellsdielbh589induziertedapkaktivierungfordertapoptoseundautophagieinhumanendickdarmkrebszellen AT gandesirimuktheshwar dielbh589induziertedapkaktivierungfordertapoptoseundautophagieinhumanendickdarmkrebszellen |