Publications
Detailed Information
Anticancer effect of LPS and IFN- γ pretreated canine macrophage-derived extracellular vesicles through apoptosis in canine melanoma and osteosarcoma cell lines : 개 흑색종 및 골육종 세포주에서 LPS와 IFN-γ 전처리한 개 대식세포 유래 세포외 소포의 세포자멸사를 통한 항암 효과
DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | 윤화영 | - |
dc.contributor.author | 이경미 | - |
dc.date.accessioned | 2020-10-13T03:40:27Z | - |
dc.date.available | 2020-10-13T03:40:27Z | - |
dc.date.issued | 2020 | - |
dc.identifier.other | 000000163213 | - |
dc.identifier.uri | https://hdl.handle.net/10371/170276 | - |
dc.identifier.uri | http://dcollection.snu.ac.kr/common/orgView/000000163213 | ko_KR |
dc.description | 학위논문 (석사) -- 서울대학교 대학원 : 수의과대학 수의학과, 2020. 8. 윤화영. | - |
dc.description.abstract | Extracellular vesicles (EVs) are particles naturally released from cells and
represent an endogenous mechanism for intercellular communication involving the transport of functionally active biological molecules. In addition to their physiological role, EVs have recently attracted interest as vehicles for delivering therapeutic molecules. The purpose of this study was to evaluate the effect of EV derived from canine M1-polarized macrophages (M1EVs) on canine tumor cells, such as D17 (osteosarcoma cells) and LMeC (melanoma cells), and to determine their effects on immune cells around the tumor. By pretreatment with LPS and IFN-γ, macrophages were polarized to M1 type. In the case of M1EV, the contents of pro-inflammatory cytokines such as TNF-α, IL-6 and IL-1β was increased, and nitrate/nitrite contents were also increased. M1EV induced apoptosis of tumor cells by increasing caspase-3 and caspase-7 activation. In addition, M1EVs decreased expression of CCR4, Foxp3 and CTLA-4 in canine peripheral mononuclear cells cocultured with tumor cells. These results suggest that use of M1EV could be an effective anti-cancer therapeutic approach in melanoma and osteosarcoma and M1EVs can be used as immunomodulators in the tumor microenvironment for cancer treatment. | - |
dc.description.abstract | 세포외 소포는 세포로부터 자연적으로 방출된 입자이며 기능적으로
활성화된 생물학적 분자에 대한 수송을 포함한 세포 간 커뮤니케이션을 위한 내인성 메커니즘을 나타낸다. 이러한 생물학적 기능뿐만 아니라, 세포외 소포는 최근 치료 분자를 전달하는 매개체로서 관심을 끌고 있다. 이 연구의 목표는 1형으로 분극된 개 대식세포주 (DH82 cell line)로부터 얻은 세포외 소포(M1EV)의 항암 효과를 개 흑색종 세포주 (LMeC cell line)와 개 골육종 세포주 (D17 cell line)를 통해 평가하고, 주변의 면역 세포에 미치는 영향을 결정하는 것이였다. LPS와 IFN- γ를 전처치하여 DH82 세포를 제1형으로 분극화한 후 세포외 소포를 획득하였다. M1EV 내에 함유된 TNF-α, IL-6 and IL-1β와 같은 전염증성 사이토카인이 증가하였고, nitrate/nitrite 함유량도 또한 증가하였다. 또한 M1EV는 caspase-3와 caspase-7 활성을 증가시킴으로써 종양 세포의 세포자멸사를 유도하였다. 그리고 개 말초 단핵세포와 종양 세포를 공배양하였을 때, M1EV는 개 말초 단핵 세포의 CCR4, Foxp3, 그리고 CTLA-4 발현을 감소시켰다. 이러한 결과는 M1EV가 흑색종 및 골육종에 대한 효과적인 항암 치료 접근법일 수 있고, M1EV가 종양 치료에 있어서 종양 미세환경 내에서의 면역 조절 역할을 할 수 있음을 시사한다. | - |
dc.description.tableofcontents | 1. Introduction‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧1
2. Materials and Methods‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧4 2.1. Cell culture 2.2. Isolation of canine peripheral blood mononuclear cells (cPBMCs) 2.3. Stimulation of M1 polarization with LPS and IFN- γ 2.4. Isolation and characterization of canine macrophage-derived extracellular vesicles (M-EVs) 2.5. Cell viability assay 2.6. RNA extraction, cDNA synthesis, and real-time quantitative PCR (RTqPCR) 2.7. Coculture of cPBMCs with D17 and LMeC cell lines 2.8. Griess assay 2.9. Flow cytometry analysis 2.10. Western blot analysis 2.11. Statistical analysis 3. Results‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧11 3.1. LPS and IFN- γ promote M1 macrophage polarization 3.2. EVs derived from macrophage stimulated with LPS and IFN- γ increase the expression of inflammatory cytokines 3.3. Concentrations of nitrite and nitrate increase in M-EVs derived from M1-polarized macrophages 3.4. M-EVs have a cytotoxic effect on the osteosarcoma and melanoma cell lines 3.5. M-EVs upregulate the protein levels of caspase-3 and caspase-7 in the osteosarcoma and melanoma cell lines 3.6. M-EVs increase apoptosis in the osteosarcoma and melanoma cell lines 3.7. M-EVs modulate CTLA-4, CCR4, and Foxp3 expressions in cPBMCs cocultured with tumor cell lines 4. Discussion‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧17 5. Conclusion‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧22 6. References‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧23 국문초록‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧42 | - |
dc.language.iso | eng | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject | Extracellular vesicle | - |
dc.subject | macrophage | - |
dc.subject | anticancer agent | - |
dc.subject | melanoma | - |
dc.subject | osteosarcoma | - |
dc.subject | apoptosis | - |
dc.subject | 세포외 소포 | - |
dc.subject | 대식구 | - |
dc.subject | 항암제 | - |
dc.subject | 흑색종 | - |
dc.subject | 골육종 | - |
dc.subject | 세포자멸사 | - |
dc.subject.ddc | 636.089 | - |
dc.title | Anticancer effect of LPS and IFN- γ pretreated canine macrophage-derived extracellular vesicles through apoptosis in canine melanoma and osteosarcoma cell lines | - |
dc.title.alternative | 개 흑색종 및 골육종 세포주에서 LPS와 IFN-γ 전처리한 개 대식세포 유래 세포외 소포의 세포자멸사를 통한 항암 효과 | - |
dc.type | Thesis | - |
dc.type | Dissertation | - |
dc.contributor.department | 수의과대학 수의학과 | - |
dc.description.degree | Master | - |
dc.date.awarded | 2020-08 | - |
dc.contributor.major | 임상수의학 | - |
dc.identifier.uci | I804:11032-000000163213 | - |
dc.identifier.holdings | 000000000043▲000000000048▲000000163213▲ | - |
- Appears in Collections:
- Files in This Item:
Item View & Download Count
Items in S-Space are protected by copyright, with all rights reserved, unless otherwise indicated.