초록 ▼

여러 종류의 전통식품에 함유된 항염증 및 암예방 물질들을 대상으로 정상세포주, 암세포주, 마우스 모델을 이용하여 발암억제 효능 및 그 기전을 규명함으로써 우수한 암 예방제의 후보물질을 도출하였음. 인체유방 상피세포주인 MCF-10A에서 브로콜리의 활성성분인 설포라펜이 NF-kB의 활성 억제를 통해 발암과정에 관여하는 COX-2의 발현을 억제함을 규명함. 내인성 발암물질인 TNF-a에 의해 정상 MCF-10A세포 전이 기전을 규명하고, 이를 사과에 함유되어 있는 phloretin이 억제함을 분자적 수준에서 규명함. 발암촉진물질인 TP

여러 종류의 전통식품에 함유된 항염증 및 암예방 물질들을 대상으로 정상세포주, 암세포주, 마우스 모델을 이용하여 발암억제 효능 및 그 기전을 규명함으로써 우수한 암 예방제의 후보물질을 도출하였음. 인체유방 상피세포주인 MCF-10A에서 브로콜리의 활성성분인 설포라펜이 NF-kB의 활성 억제를 통해 발암과정에 관여하는 COX-2의 발현을 억제함을 규명함. 내인성 발암물질인 TNF-a에 의해 정상 MCF-10A세포 전이 기전을 규명하고, 이를 사과에 함유되어 있는 phloretin이 억제함을 분자적 수준에서 규명함. 발암촉진물질인 TPA에 의해 유도된 마우스 피부암을 phloretin과 메밀의 주성분인 rutin이 현저히 억제하며, COX-2의 발현을 억제함을 확인함. 무모마우스를 이용하여 UVB에 의한 COX-2의 발현기전을 규명하고, rutin에 의한 억제 기전을 규명함. 또한 대장암과 전립선암세포를 사용하여 17종의 전통식품 성분 중 암세포 증식 억제 효능이 뛰어난 phloretin, fucoidan, 3,3'-didindoylymethane (DIM)을 선별하였고, 이들이 대장암과 전립선암세포의 세포사멸을 미토콘드리아 경로를 통해 유도함을 체계적으로 규명함. 본 연구 결과는 효능이 탁월한 암예방 후보물질 물질 도출에 대한 근간이 되었고, 17여개의 국내외 SCI급 논문 및 특허를 통해 연구결과를 발표하였음.

Abstract ▼

We here report that sulforaphane (SFN), an isothiocyanate found in cruciferous vegetables, inhibited the cyclooxygenase-2 (COX-2) expression in human mammary epithelial cells (MCF10A) treated with tumor promoter TPA. TPA-induced COX-2 expression is associated with nuclear factor-kappa B (NF-κB) act

We here report that sulforaphane (SFN), an isothiocyanate found in cruciferous vegetables, inhibited the cyclooxygenase-2 (COX-2) expression in human mammary epithelial cells (MCF10A) treated with tumor promoter TPA. TPA-induced COX-2 expression is associated with nuclear factor-kappa B (NF-κB) activation via catalytic activation and phosphorylation of ERK1/2 and p38 MAPK in MCF10A cells. SFN inhibited the NF-κB DNA binding activity and transcriptional activity via suppression of ERK1/2, but not affect on p38 MAPK. We also observed that endogenous tumor promoter TNF-α induced COX-2 expression in MCF10A cells which is abolished by genestein. TNF-α induced invasiveness of MCF10A cells via production of urokinase plasminogen activator (uPA). TNF-α also induced expression and DNA binding activity of β-catenin which act as a potential link between inflammation and cancer. TNF-α induced expression of uPA is mediated by activation of β-catenin in MCF10A cells through generation of reactive oxygen species. Phloretin, which is present in apples and pears, inhibited expression of COX-2 in MCF10A cells treated with TNF-α. Phloretin also inhibited uPA expression and expression and transcriptional activity of β-catenin via blocking generation of ROS. In addition, phloretin and rutin inhibited number and incidence of tumor in DMBA/TPA-induced mouse skin carcinogenesis. Phloretin inhibited TPA-induced COX-2 expression in a concentration dependent manner. Phloretin also inhibited the catalytic activity and phosphorylation of ERK1/2. Moreover, phloretin inhibited DNA binding activity of NF-κB and phosphorylation of IκB-α and its translocation into the nucleus in mouse skin treated with TPA. UVB, a major skin carcinogen, induced expression of COX-2 and iNOS in HR-1 hairless mouse. UVB also activated the DNA binding activity of NF-κB and IκB kinase β (IKKβ) thereby induced phosphorylation and tranlocation into the nucleus of IκB-α, p65, and p50. UVB activated the ERK1/2 and p38MAPK by phosphorylation. We also observed that a pharmacological specific IKKβ inhibitor SC-515 attenuated the expression of iNOS but not COX-2 induced by UVB in HR-1 hairless mouse. Rutin inhibited the expression of COX-2 as determined by Western blot analysis and immunohistochemisty analysis. Rutin inhibited the epxresion of 4-HNE, phosphorylation of p38 MAPK, c-jun-NH2 terminal kinase and DNA binding activity of activator protein-1 in HR-a hairless mouse treated with UVA. We also identify the most potent compound among the traditional food constituents such as cyanidine, deplhindine, hesperidin, apigenin, puerarin, essential oil, eugenol, rosmarinic acid, fucoidan, phloretin, indole-3-carbinol, 3,3'-diindolymethane (DIM), genistein, daidzein, piceatannol, and resveratrol using colon cancer cells (HT-29, HCT116) and prostate cancer (DU145, LNCap) cells as determined by MTT assay. Among the above compounds, phloretin and fucoidan strongly inhibited the growth of HT-29, whereas DIM inhibited the growth of HT-29 and DU145 cells. Phloretin induced proteolytic cleavage of caspase-8, -9, -3, and -7 and its substrate PARP and increased ration of Bax/Bcl-2 in a concentration dependent manner in HT-29 cells. Phloretin also release of cytochrome C and expression of Smac/Diable in the cytosolic fraction of HT-29 cells. Focoidan induced apoptotic cell population and increased pro-apoptotic proteins such as Bak and Bad and decreased expression of Mcl-1. Focoidan induced apoptotic cell population and increased pro-apoptotic proteins such as Bak and Bad and decreased expression of Mcl-1. Focoidan also disrupted the mitochondrial membrane potential in HT-29 cells. Fucoidan also induced proteolytic cleavage of caspase-8,-9,-7,-3 and PARP in HT-29 cells. Moreover, fucoidan disrupted the mitochomdrial membrane pototentials and inhibited the expression of inhibitor of apoptosis proteins (IAP). Pharmacological caspase-8 inhibitor Z-IETD-FMK attenuated the focoidan-induced apoptosis in HT-29 cells, suggesting that focoidan induced apoptosis via death receptor- and mitochondria-mediated pathway. DIM induced apoptosis in colon cancer cells and prostate cancer cells. DIM increased expression of Fas, proteolytic cleavage of caspase-8, and truncated-Bid in both cells. Expression of Bax is increased in prostate cancer cells, but not in colon cancer cells. DIM released the cytochrome c and Smac/Diablo into the cytoplasm and induced proteolytic cleavage of caspase-9,-7, and -3 in both cells. The caspase-8 inhibitor abolished the DIM-induced cell death.

목차 Contents

• 제 1 장 연구개발과제의 개요...10
• 제1절 연구개발의 목적...10
• 제2절 연구개발의 필요성...11
• 제3절 연구개발의 범위...12
• 제 2 장 국내외 기술개발 현황...13
• 제 3 장 연구개발수행 내용 및 결과...14
• 제1절 이론적. 실험적 접근방법...14
• 제2절 연구수행 결과...16
• 제 4 장 목표달성도 및 관련분야에의 기여도...63
• 제 5 장 연구개발결과의 활용계획...68
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보...69
• 제 7 장 참고문헌...70