초록 ▼

본 연구과제는 Arsenic trioxide(As2O3)가 만성 골수성 백혈병(CML) 세포주에서의 cytotoxicity와 apoptosis에 미치는 영향을 조사하고, Bcr-Abl 발현 양상에 미치는 영향을 관찰하며, apoptosis와 관련된 작용기전에 대해 규명함으로써 만성 골수성 백 혈병의 새로운 치료제로서의 토대를 마련하고자 하였다.
1. Arsenic trioxide(As2O3)가 CML 세포주의 세포독성 및 apoptosis에 미치는 영향
- 세포독성능 측정
- DNA 분절 측정
- Hoe

본 연구과제는 Arsenic trioxide(As2O3)가 만성 골수성 백혈병(CML) 세포주에서의 cytotoxicity와 apoptosis에 미치는 영향을 조사하고, Bcr-Abl 발현 양상에 미치는 영향을 관찰하며, apoptosis와 관련된 작용기전에 대해 규명함으로써 만성 골수성 백 혈병의 새로운 치료제로서의 토대를 마련하고자 하였다.
1. Arsenic trioxide(As2O3)가 CML 세포주의 세포독성 및 apoptosis에 미치는 영향
- 세포독성능 측정
- DNA 분절 측정
- Hoechst 33258 염색을 통한 핵 형태 변화 관찰
- 전자현미경을 통한 세포 형태 관찰
- Caspase-3의 활성화
- 세포주기 분석
2. Bcr-abl expression에 관한 연구
- RT-PCR에 의한 bcr-abl 융합유전자 발현의 변화 관찰
- Western blotting에 의한 Bcr-abl 융합단백질 발현의 변화 관찰
3. Apoptosis와 관련된 MAP kinase의 신호전달과정에 관한 연구
- Western blotting을 통한 ERK1/2, p38, JNK 등의 활성화
- Inhibitor 처리 후의 변화

Abstract ▼

Through examining the effects of Arsenic trioxide(AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis,) on the apoptosis, Bcr-Abl expression and mitogen-activated protein(MAP) kinases signaling pathway in chronic myelogenous leukemia(CML) cells, this study aims at the elucidat

Through examining the effects of Arsenic trioxide(AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis,) on the apoptosis, Bcr-Abl expression and mitogen-activated protein(MAP) kinases signaling pathway in chronic myelogenous leukemia(CML) cells, this study aims at the elucidation of intracellular signal mechanism and application to the therapy of CML 1. Effect of Arsenic trioxide(AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis,) on cytotoxicity and apoptosis - Cytotoxicity - DNA fragmentation - Morphological stududies - Activation of Caspase-3 - Cell cycle analysis 2. Effects on Bcr-Abl expression - Effect of AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, on Bcr-Abl gene expression by RT-PCR - Effect of AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, on Bcr-Abl fusion protein by Western blotting 3. Effects on the activities of MAP kinases - Activations of ERK1/2, p38, JNK proteins To determine if AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, might be useful for treatment of CML, it was examined the ability of AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, to induce apoptosis in K562 cells. As the result of cytotoxicity, the IC50 value for AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, was to be 10 μM. DNA fragmentation became evident after the incubation of the cells with 20 μM of AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, for 12 h and 24 h, respectively. Morphological changes and chromatin condensation of the cells undergoing apoptosis were also observed. After treatment, activation of caspase-3 was observed 12 h after the start of treatment with 20 M by Western blot analysis. And DNA flow cytometric analysis indicated that AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, induced G2/M arrest in these cells. And the treatment of AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, downregulated not only the expression of Bcr-Abl oncoprotein but also bcr-abl mRNA levels, suggesting a transcriptional regulation. Next, the relationship between MAP kinase response in K562 cells treated with AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, was investigated. AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, at 10 μM strongly induced the activation of p38, and JNK 1/2, while ERK 1/2 were inhibited. MEK 3/6, which acts upstream of p38, and ATF-2, a transcription factor, were also activated after treatment of AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis,. In addition, the pretreatment of SB203580, a specific inhibitor of p38, inhibited AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, induced apoptotic cell death. These results suggest that AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, is able to induce the apoptotic activity in K562 cells and its apoptotic mechanism may be associated with down regulation of Bcr-Abl, the activation of caspase-3 and p38 kinase. It suggests that AThrough examining the effects of Arsenic trioxide(As2O3) on the apoptosis, may be a potential agent for treatment of CML.

목차 Contents

• Ⅰ. 연구계획 요약문...3
• 1. 국문요약문...3
• Ⅱ. 연구결과 요약문...4
• 1. 국문요약문...4
• 2. 영문요약문...5
• Ⅲ. 연구내용 및 결과...6
• 1. 서론...6
• 2. 연구방법 및 이론...6
• 3. 결과 및 고찰...10
• 4. 결론...15
• 5. 인용문헌...16