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JAK/STAT 인산화 및 VEGFR-2/Akt/mTOR 신호전달 억제를 통한 독사조신의 난소암 억제 기전 규명
DC Field | Value | Language |
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dc.contributor.advisor | 김대용 | - |
dc.contributor.author | 박미선 | - |
dc.date.accessioned | 2017-07-13T16:43:20Z | - |
dc.date.available | 2017-07-13T16:43:20Z | - |
dc.date.issued | 2015-02 | - |
dc.identifier.other | 000000025572 | - |
dc.identifier.uri | https://hdl.handle.net/10371/120207 | - |
dc.description | 학위논문 (박사)-- 서울대학교 대학원 : 수의학과, 2015. 2. 김대용. | - |
dc.description.abstract | Doxazosin, a commonly prescribed treatment for patients with benign prostatic hyperplasia, serves as an α1-blocker of the adrenergic receptors. In this study, we calculated its effect on the ovarian carcinoma cell system and animal model. Doxazosin induces dose-dependent growth and time-dependent suppression and is additively activated through IFN-α or IFN-γ stimulation. At the same time, they both enhanced G1 phase arrest, as well as the activity of caspase-3 and PARP, and the reduction of cyclin D1 and CDK4 protein levels. Doxazosin growth suppression was abolished either by the Janus family of tyrosine kinase (JAK) or the signal transducer and activator of transcription (STAT) inhibitor treatment. The activity of JAK/STAT was dependent on the level of doxazosin, suggesting a requirement of doxazosin for the activation of JAK/STAT. Furthermore, doxazosin plus IFN-α or doxazosin plus IFN-γ additively suppressed the activation of the JAK/STAT signals through phosphorylation of JAK and STAT, thus affecting the activation of subsequent downstream signaling components PI3K, mTOR, 70S6K and PKCδ. And in vivo study demonstrated that doxazosin significantly suppressed tumor growth in an ovarian xenograft model, inducing apoptotic cell death by up-regulating the expression of p53, whereas c-Myc expression was markedly reduced. Therefore, our data findings indicate that doxazosin can modulate the apoptotic effects of IFN-α and IFN-γ through the JAK/STAT signaling pathways. Collectively, we indicate that this action may be a potent chemotherapeutic property against ovarian carcinoma. Doxazosin also inhibited vascular endothelial growth factor (VEGF)-induced HUVEC migration as well as capillary-like structure tube formation in vitro. It also suppresses the expression of HIF-1α and VEGF in ovarian carcinoma cells. Doxazosin inhibited phosphorylation of vascular endothelial growth factor receptor-2 (VEGFR-2) and transcription of VEGFR-2. Doxazosin inhibited PI3K, Akt, PDK1 and mTOR phosphorylation but had no effect on ERK1/2 phosphorylation by VEGF treatment. Our results provide evidence for the cellular function in endothelial cell system that is relevant to angiogenesis through the inhibition of the Akt/mTOR phosphorylation by interacting with VEGFR-2. Furthermore, doxazosin prohibited VEGFR-2 phosphorylation and suppressed tumor vascularization in a xenograft model of human ovarian cancer. We found the biological function of doxazosin to be a potent anti-ovarian cancer agent by inhibition of JAK/STAT phosphorylation and anti-angiogenic agent by suppression of VEGFR-2 signaling pathway. Therefore, doxazosin combination therapies may be a more useful approach for more advanced ovarian cancers and recurrent patients | - |
dc.description.tableofcontents | ABSTRACT i
CONTENTS iv ABBREVIATION 1 LITERATURE REVIEW 2 Ovarian cancer chemotherapy 2 Interferons and JAK/STAT pathway 10 VEGFR-2/AKT/mTOR pathway 13 Doxazosin 16 Objective 17 CHAPTER I DOXAZOSIN SUPPRESSES JAK/STATs PHOSPHORYLATION THROUGH IFN-α/γ INDUCED APOPTOSIS 18 Abstract 19 Introduction 21 Materials and Methods 25 Cell lines, drug reagents and antibodies 25 Cytotoxicity assay 26 Cell cycle analysis 26 Western blotting 27 Xenograft mouse model 27 Statistical analysis 28 Results 29 Doxazosin inhibits cell proliferation in a dose-dependent manner, and additively enhances apoptotic cell death by IFN-α and IFN-γ treatment 29 Effect of doxazosin on IFN-α or IFN-γ-stimulated expression of cell cycle modulator proteins 38 Doxazosin inhibits phosphorylation of JAK/STAT 43 Decreased phosphorylation of PI3K/Akt/mTOR as well as 70S6K and PKCδfollowing doxazosin treatment 46 Doxazosin inhibits tumor growth in nude mice 50 Discussion 57 CHAPTER II DOXAZOSIN INHIBITS TUMOR GROWTH AND ANGIOGENESIS BY DECREASING VEGFR-2/Akt/mTOR SIGNALING AND VEGF AND HIF-1α EXPRESSION 62 Abstract 63 Introduction 64 Materials and Methods 68 Cell lines, drug reagents and antibodies 68 [3H] thymidine incorporation analysis 69 Cell migration analysis 69 In vitro tube formation analysis 70 PI3K activity analysis 71 Western blotting 72 Xenograft mouse model 72 Immunohistochemistry 73 Statistical analysis 74 Results 75 Doxazosin significantly suppressed VEGF-induced cell proliferation, migration, and capillary-like tubule formation in HUVECs 75 Doxazosin suppresses PI3K and Akt phosphorylation in a concentration-dependent manner 80 Doxazosin inhibits VEGF-induced VEGFR-2 phosphory lation and VEGFR-2-dependent transcription 93 Doxazosin inhibits tumor growth by antiangiogenic activity in vivo 96 Discussion 101 GENAERAL DISCUSSION AND CONCLUSION 104 ABSTRACTS IN KOREAN 108 REFERENCES 110 감사의 글 121 | - |
dc.format | application/pdf | - |
dc.format.extent | 2555058 bytes | - |
dc.format.medium | application/pdf | - |
dc.language.iso | ko | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject | doxazosin | - |
dc.subject | ovarian cancer | - |
dc.subject | IFN | - |
dc.subject | JAK | - |
dc.subject | STAT | - |
dc.subject | VEGFR-2 | - |
dc.subject | Akt | - |
dc.subject | mTOR | - |
dc.subject.ddc | 636 | - |
dc.title | JAK/STAT 인산화 및 VEGFR-2/Akt/mTOR 신호전달 억제를 통한 독사조신의 난소암 억제 기전 규명 | - |
dc.type | Thesis | - |
dc.description.degree | Doctor | - |
dc.citation.pages | vii, 121 | - |
dc.contributor.affiliation | 수의과대학 수의학과 | - |
dc.date.awarded | 2015-02 | - |
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