인체거대세포 바이러스에 감염된 세포에서의 미토콘드리아 경유 세포사 = Mitochondria Targeted Apoptosis in Human Cytomegalovirus Infected Cells

다국어 초록 (Multilingual Abstract)

Mitochondria often play a central role in an apoptotic pathway. Disruption of mitochondrial transmembrane potential (ΔΨm) has been observed in various cells undergoing apoptosis. HCMV infection induces apoptosis in permissive cells, however, there was not many investigations about mitochondria targeted apoptosis in HCMV infected human foreskin fibroblast (HFF) cells. Here, we studied the mitochondrial pathway of apoptosis in HCMV infected HFF cells. Flow cytometry analysis using JC-1 revealed that HCMV infection induced disruption of ΔΨm in HFF cells which started at 24 hours post infection (h.p.i) and maximized at 48 h.p.i. Cytochrome c (cyt-c), an inner membrane protein of mitochondria, was detected in cytoplasmic extracts of the HCMV infected cells by western blot analysis after 24 h.p.i., but not of mock infected cells, suggesting that ΔΨm was disrupted and cyt-c was released from mitochondria to cytoplasm. Caspase activity assay based on fluorescence spectrophotometery using fluorogenic substrate revealed that caspase-3 activity was increased after 48 h.p.i. in HCMV infected cells. Caspase-8 activity was increased after 72 h.p.i. in HCMV infected cells. Therefore, HCMV infection induced mitochondria targeted apoptosis in HFF cells.

목차 (Table of Contents)

• I. 서론 1
• II. 재료 및 연구방법 4
• 1. 세포배양 4
• 2. 바이러스 4
• 3. HCMV 감염과 caspase-3, -8 저해제 처리 5
• 4. Propidium Iodide Staining 6
• 5. JC-1 staining 6
• 6. Western blot 분석 7
• 7. Caspase activity analysis 8
• Ⅲ. 결과 9
• 1. HCMV 감염에 의한 HFF 세포의 apoptosis 유도 9
• 2. HCMV 감염에 의한 HFF 세포의 미토콘드리아 막 전위 변화 9
• 3. HCMV의 감염에 의한 cytochrome c 방출 12
• 4. HCMV 감염에 의한 caspase-3 활성 16
• 5. HCMV 감염에 의한 caspase-8 활성 16
• 6. Caspase 저해제 처리에 의한 caspase의 활성 측정 19
• Ⅳ. 고찰 22
• Ⅴ. 결론 25
• Ⅵ. 참고문헌 26
• Ⅶ. 요약 30