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암세포에서 bortezomib과 nutlin-3 병합처리를 통한 paraptosis 유도 기전 연구
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 최경숙 | - |
| dc.contributor.author | Lee Dong Min | - |
| dc.date.accessioned | 2019-10-21T07:27:13Z | - |
| dc.date.available | 2019-10-21T07:27:13Z | - |
| dc.date.issued | 2016-02 | - |
| dc.identifier.other | 22078 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/18820 | - |
| dc.description | 학위논문(석사)--아주대학교 일반대학원 :의생명과학과,2016. 2 | - |
| dc.description.tableofcontents | I. INTRODUCTION 1 II. MATERIALS AND METHODS 7 A. Chemicals and antibodies 7 B. Cell culture of various cancer cell lines 7 C. Examination of the stable cell lines expressing the fluorescence specifically in mitochondria or endoplasmic reticulum 8 D. Measurement of cellular viability 8 E. MTT assay 9 F. Western blotting 9 G. Immunocytochemistry 9 H. Measurement of cytosolic and mitochondrial Ca² levels 10 I. Small interfering RNAs 10 J. shRNA-mediated knockdown of proteins 10 K. Transmission electron microscopy 11 L. Statistical analysis 11 M. Isobologram analysis 11 III. RESULTS 13 1. Bortezomib and nutlin-3 synergistically induce cell death in various cancer cell lines with defective p53 13 2. Vacuolation induced by bortezomib plus nutlin-3 is derived from the dilation of both the ER and mitochondria 27 3. Protein synthesis is required for vacuolation and subsequent cell death by bortezomib plus nutlin-3 33 4. CHOP induction critically contributes to the ER dilation and the cell death by bortezomib plus nultin-3 36 5. Mitochondria Ca2+ uniporter-mediated mitochondrial Ca² overload is important for the cell death induced by bortezomib plus nutlin-3 41 IV. DISCUSSION 50 V. REFERENCES 68 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | 암세포에서 bortezomib과 nutlin-3 병합처리를 통한 paraptosis 유도 기전 연구 | - |
| dc.title.alternative | Nutlin-3 sensitizes cancer cells to bortezomib via induction of paraptosis-associated cell death | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | Dong Min Lee | - |
| dc.contributor.department | 일반대학원 의생명과학과 | - |
| dc.date.awarded | 2016. 2 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 739586 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000022078 | - |
| dc.subject.keyword | bortezomib | - |
| dc.subject.keyword | nutlin-3 | - |
| dc.subject.keyword | cancer | - |
| dc.subject.keyword | paraptosis | - |
| dc.subject.keyword | endoplasmic reticulum | - |
| dc.subject.keyword | mitochondria | - |
| dc.description.alternativeAbstract | The ubiquitin-proteasome system is a complex and tightly controlled system in charge of degrading 80-90% of proteins, and is central to regulating cellular function and keeping protein homeostasis. Since proteasome inhibition can lead to the accumulation of the misfolded proteins and subsequent proteotoxicity in tumor cells, the proteasome is considered a target for cancer therapies. Bortezomib, the first FAD approved proteasome inhibitor, is now in the clinic to effectively treat multiple myeloma and mantle cell lymphoma, but its efficacy in solid tumors is not satisfactory. In this study, we show that the nutlin-3, a small-molecule inhibitor of murine double minute 2 (MDM2), effectively overcomes the resistance of MDA-MB 435S breast cancer cells to bortezomib. While bortezomib treatment did not markedly alter cellular morphologies, nutlin-3 treatment induced mitochondrial dilation without induction of cell death. Alternatively, combined treatment with bortezomib and nutlin-3 effectively induced cell death, which was accompanied by the simultaneous dilation of mitochondria and the endoplasmic reticulum (ER). These results suggest that nutlin-3 may overcome the resistance of these breast cancer cells to bortezomib via induction of paraptosis-like cell death. We found that the combined treatment yielded in MCU channeled calcium increase. This is supported with inhibition of MCU by Ru360 or knockdown of MCU significantly attenuated the cell death by the combined treatment. We also found that nutlin-3 co-treatment remarkably potentiated the bortezomib-mediated upregulation of ER marker proteins, including ATF4 and CHOP. Knockdown of CHOP effectively inhibited the cellular vacuolation and subsequent cell death by the combined treatment, suggesting that upreugulation of CHOP plays an important role in this process. Taken together, our results suggest that MCU-mediated mitochondrial Ca²⁺ overload and CHOP-mediated ER dilation may critically contribute to paraptosis-like cell death induced by the combination of bortezomib and nutlin-3 in breast cancer cells. | - |
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