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Apoptotic cell death of human colon cancer cell lines after extracellular calgranulin B uptake : 대장암세포주의 외래 Calgranulin B 단백질 포식에 의한 세포사멸

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dc.contributor.advisor구자록-
dc.contributor.author김근-
dc.date.accessioned2017-07-14T01:15:28Z-
dc.date.available2017-07-14T01:15:28Z-
dc.date.issued2015-08-
dc.identifier.other000000066656-
dc.identifier.urihttps://hdl.handle.net/10371/121774-
dc.description학위논문 (박사)-- 서울대학교 대학원 : 협동과정 종양생물학전공, 2015. 8. 구자록.-
dc.description.abstractThe molecular mechanisms through which extracellular calgranulin B affects colon cancer cells remain unclear. My present study focused on the systemic link between calgranulin B and colon cancer. In patients tumor tissues, calgranulin B protein showed a positive correlation with stromal inflammatory cells surrounding tumor cells, and methylation of the gDNA promoter region of calgranulin B was observed in both patients paired tissues and human colon cancer cell lines. Notably, extracellular calgranulin B could penetrate colon cancer cells, but not the other types of cancer cells tested. Internalization of calgranulin B was possible via various mechanisms depending on colon cancer cells and caused apoptotic cell death. In addition, calgranulin B inhibited aurora A kinase activity in a dose-dependent manner and suppressed tumor growth in a mouse model. My findings demonstrate the antitumor effects of calgranulin B in the inflammatory microenvironment of colon cancer.-
dc.description.tableofcontentsAbstract i

Contents iii

List of tables vi

List of figures vii

Introduction 1

Materials and Methods 9
Human cancer cell lines 9
Western blot analysis 9
gDNA PCR 10
Bisulfite-modified sequencing analysis 11
Immunohistochemistry of tumor tissues and statistical analysis 12
Confocal microscopy 14
Flow cytometry 15
3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay 15
Cell cycle analysis 16
Terminal transferase dUTP nick end labelling assay 17
In vivo study 17
18F-FDG positron emission tomography?computed tomography imaging 18
Immunohistochemistry in the xenograft model 19
Construction of Glutathione S-Transferase (GST)-human calgranulin B fusion protein 19
Human protein microarray 21
Immunoprecipitation 21
Aurora A kinase assay 22

Results 24
Calgranulin B was not expressed in colon cancer cell lines. 24
Tissue calgranulin B level in tumor cells was correlated with that of stromal inflammatory cells around tumor glands in colon cancer. 29
Internalization of extracellular calgranulin B into colon cancer cells was mediated by various pathways depending on cell type. 32
Extracellular treatment of calgranulin B induced antitumor effects in colon cancer cells. 37
Intravenous injection of calgranulin B suppressed tumor volume in the HCT-116 colon cancer xenograft model. 42
Calgranulin B interacted with aurora A kinase and the activity of aurora A kinase decreased in the presence of calgranulin B. 45

Discussion 54

References 62

Abstract in Korean 77
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dc.formatapplication/pdf-
dc.format.extent2082424 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 대학원-
dc.subjectcalgranulin B-
dc.subjectcolon cancer-
dc.subjectprotein internalization-
dc.subjectinflammatory microenvironment-
dc.subjectaurora A kinase-
dc.subject.ddc616-
dc.titleApoptotic cell death of human colon cancer cell lines after extracellular calgranulin B uptake-
dc.title.alternative대장암세포주의 외래 Calgranulin B 단백질 포식에 의한 세포사멸-
dc.typeThesis-
dc.contributor.AlternativeAuthorKun Kim-
dc.description.degreeDoctor-
dc.citation.pagesvii, 79-
dc.contributor.affiliation의과대학 협동과정 종양생물학전공-
dc.date.awarded2015-08-
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