인간 위암 조직의 프로테옴 분석 : 암조직에서 Rho GDP dissociation inhibitor-1,-2의 과발현 /d金유娜 [저]

다국어 초록 (Multilingual Abstract)

Gastric cancer remains a leading cause of cancer-related death worldwide. Advances in diagnostic and treatment technologies have enabled us to offer excellent long-term survival results for early gastric cancer, but prognosis of advanced gastric cancer still remains poor. The aim of this work was to figure out the protein components that are differentially expressed in stomach cancer tissues to develop the protein candidates for tumor marker. In this study, we have analyzed the proteomes of 152 stomach cancer tissue samples and compared with those of noncancerous tissues of the same patients. Human stomach tissue samples were prepared from resection materials of gastric cancer patients. The proteins of stomach tissues were analyzed by two-dimensional electrophoresis, stained with silver nitrate and the images were analyzed with the aid of PDQuest^(TM). Of the 152 samples that gave readable gel image, 11 samples were positive for overexpression (with a cutoff value of 3.0) of the spot 3211 and 17 samples were positive for overexpression of the spot 3233. The spot was excised from preparative gels, digested by trypsin, and subjected to peptide mass fingerprinting. The spot was identified to be Rho GDP dissociation inhibitor 1 and 2, with the sequence coverage of 28% and 29%, respectively. In the previous studies reported, RhoGDI has been associated with tumor cell invasion in ovarian cancer and RhoGDI was overexpressed in breast cancer. We first found that RhoGDI-1 was significantly overexpressed in human stomach cancer.

목차 (Table of Contents)

• 목차 =ⅰ
• LIST OF FIGURES = ⅳ
• LIST OF TABLES = ⅵ
• ABSTRACT = ⅶ
• Ⅰ. 서론 = 1
• Ⅱ. 실험재료 및 방법 =7
• 1. 시약 및 재료 = 7
• 2. 위 조직 시료 = 8
• 3. 실험 방법 = 8
• 1) 단백질 시료 준비 = 8
• 2) 단백질 정량 = 9
• 3) 이차원 전기영동 = 10
• (1) Rehydration of IPG gel strip = 10
• (2) Isoelectric Focusing (IEF) = 10
• (3) Equilibration of IPG gel strip = 10
• (4) SDS-polyacrylamide gel electrophoresis (SDS-PAGE) = 11
• 4) 염색 = 11
• (1) Silver stain = 11
• (2) Coomassie stain = 12
• 5) 이미지 분석 = 12
• 6) 단백질 동정 = 13
• (1) Excision and washing of gel pieces = 13
• (2) Reduction and alkylation = 14
• (3) In gel digestion and extraction of peptides = 14
• (4) Sample loading and MALDI-TOF-MS = 15
• (5) Search = 15
• 7) Western blottting = 16
• Ⅲ. 결과 = 17
• 1. 위 조직 단백질의 발현 양상 분석 = 17
• 2. 위암 조직과 정상 조직의 2-DE 패턴 비교 = 17
• 3. Spot 3211, spot 3233의 발현 변화 분석 = 22
• 4. 단백질 spot 3211과 spot 3233의 동정 = 31
• 5. 정상 조직과 위암 조직에서 RhoGDI-1,-2의 발현 변화 분석 = 35
• 6. Western blot 결과. = 35
• 7. RhoGDI-1,-2와 위암 환자의 임상 병리학적 특징과의 상관관계 = 40
• Ⅳ. 고찰 = 42
• Ⅴ. 결론 = 46
• Ⅵ. 참고문헌 = 47