초록 ▼

-연구개발결과
각막 keratocyte 세포주에서 cytokine에 의한 phenotypic transition과 migration을 분석한 후 RNA array를 통해 117 signal transduction genes, 301 cell proliferation과 differentiation genes, 176 apoptotic genes가 차이가 있음이 관찰됨. 각막 조직을 이용한 2D MALDI-TOF 분석 결과 Cell junction, ECM related cytoskeleton, antioxidant protein

-연구개발결과
각막 keratocyte 세포주에서 cytokine에 의한 phenotypic transition과 migration을 분석한 후 RNA array를 통해 117 signal transduction genes, 301 cell proliferation과 differentiation genes, 176 apoptotic genes가 차이가 있음이 관찰됨. 각막 조직을 이용한 2D MALDI-TOF 분석 결과 Cell junction, ECM related cytoskeleton, antioxidant protein, transcription/transformation에 관여하는 단백질의 발현량이 차이가 있음이 관찰됨.

Abstract ▼

To increase our understanding of the pathogenesis of keratoconus, we invetstigate these studies
1. Data profiling of expression genes by RNA using H2O2 induced oxidative stress in vitro model
2. cornea tissue based protein expression differences between keratoconus and normal subjects using tw

To increase our understanding of the pathogenesis of keratoconus, we invetstigate these studies
1. Data profiling of expression genes by RNA using H2O2 induced oxidative stress in vitro model
2. cornea tissue based protein expression differences between keratoconus and normal subjects using two dimensional electrophoresis (2DE) and MALDI-TOF analysis.
3. Expression Data profiling by Proteome and RNA analysis
In cell viability and apoptosis assay in H2O2 induced Cornea Epithelial and Keratocyte cell lines, we suggested that cornea epithelial are more sensitivity than the keratocyte using CCK, FACS and TUNEL assay. And we also were detected secretion of cytokines, particularly IL1 beta, in H2O2 induced Cornea Epithelial Cell line.
In the cell transition and migration in cytokine treated HK cells, we detected that it was induced more cell migration and phenotypic transition effects in IL1 beta treated keratocyte cell line.
And to construct expression profiling of keratoconus patients, we analyzed RNA and protein expression differences between keratoconus and normal subject using microarray and 2D-MALDI TOF.
In microarray Data Profiling of Human HT-12 v3.0 Expression Beadchip, we detected 1170 signal transduction genes, 301 cell proliferation and differentiation genes and 176 apoptotic genes.
And in 2D-MALDI TOF analysis, we identified 100 spots that demonstrated a significant differences between keratoconus and normal cornea with a ProtScore (>or=2). The intensity of 37 spots were markedly higher in keratoconus than normal cornea (>2 folds), and 14 spots were detected only in keratoconic cornea. Key proteins we recognized included type VI collagen alpha 2 chain, Transforming growth factor-beta-induced protein ig-h3, nebulin, spectrin, alpha-enolase, keratin, annexin, peroxiredoxin-6, type VI collagen alpha 2 chain precursor, carbonic anhydrase 1, keratin 3, 5, 14, 79, type I cytoskeletal 12 , decorin, aldehyde dehydrogenase , dystonin , transformation/transcription domain-associated protein variant. From our study, the identified proteins corresponded to molecules belonging to Cell junction, ECM related cytoskeleton, antioxidant protein and transcription/transformation. Our findings could contribute to the knowledge for the further understanding of pathogenesis of keratoconus.

목차 Contents

• 표지 ... 1
• 제출문 ... 3
• 보고서 요약서 ... 4
• 요약문 ... 5
• SUMMARY ... 6
• 6. 연구 성과 실적 및 향후 계획 ... 7
• 6.1 총괄연구개발과제의 연구성과 실적 및 향후 계획 ... 7
• 6.2 연구성과 유형별 세부 내역 ... 8
• 7. 참여연구원 현황표 ... 12
• II. 총괄연구과제 연구결과 ... 13
• 1. 연구개발과제의 배경 및 필요성 ... 14
• 2. 국내외 기술개발 현황 ... 17
• 3. 연구개발과제의 추진체계 ... 19
• 4. 연구개발수행 내용 및 결과 ... 20
• 5. 목표달성도 및 관련분야 기여도 ... 33
• 6. 향후 연구성과 추진 계획 ... 34
• 7. 연구개발결과의 파급효과 ... 34
• 8. 연구개발결과의 활용계획 ... 36
• 9. 연구개발과정에서 수집한 해외과학기술정보 ... 37
• 10. 참고문헌 ... 37
• IV. 첨부서류 ... 38
• 끝페이지 ... 39