β - Sitosterol에 의한 인체 대장암 HCT116 세포의 증식억제에 관한 연구

국문 초록 (Abstract)

β-Sitosterol은 과일과 야채 등을 포함한 대부분의 고등식물에 존재하는 중요한 phytosterol의 하나로서, 인체 암의 예방과 치료에 매우 유효한 것으로 보고되어져 오고 있다. 본 연구에서는 β-sitosterol의 암세포 증식억제 기전의 해석을 시도하기 위하여 인체 대장암세포 HCT116의 증식에 미치는 β-sitosterol의 영향을 조사하였다. β-Sitosterol의 처리로 HCT116 암세포의 증식은 처리 농도 의존적으로 감소되었으며, 특히 7.5 μM 이상 처리에서는 급격한 성장억제 효과가 있었다. 또한 5.0 μM 처리군에서부터 apoptotic body의 형성이 관찰되었고, β-catenin 단백질의 분해 현상과 연관성이 있었다. 그리고 β-sitosterol이 처리된 암세포에서는 종양억제 유전자 p53 및 Cdk inhibitor p21의 발현이 전사 및 번역 수준에서 모두 증가되었다. 본 결과는 그동안 연구가 거의 진행되어져 있지 않았던 β-sitosterol에 의한 암세포주기 조절 해석을 위한 주요한 자료로 활용될 것이다.

다국어 초록 (Multilingual Abstract)

β-Sitosterol is the major phytosterol in higher plants, including fruits and vegetables. The molecule has been shown to have the potential for prevention and therapy for human cancer. We investigated the effects of β-sitosterol on the cell proliferation of HCT116 human colon cancer cells in order to understand its anti-proliferative mechanism. β-Sitosterol treatment resulted in the inhibition of cell proliferation in a concentration-dependent manner. The anti-proliferative effect of HCT116 cells by β-sitosterol was associated with formation of apoptotic bodies and degradation of β-catenin protein. In addition, β-sitosterol-treatment induced a marked accumulation of tumor suppressor p53 and a concomitant induction of cyclin-dependent kinase (Cdk) inhibitor p21 without alteration in the levels of cyclins and Cdks. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of β-sitosterol.

목차 (Table of Contents)

• Abstract
  
• 서론
  
• 재료 및 방법
  
• 결과 및 고찰
  
• 요약
  
• 감사의 글
  
• 문헌

참고문헌 (Reference)

• 1 "β-Sitosterol inhibits growth of HT-29 human colon cancer cells by activating the sphingomyelin cycle. Anticancer Res 18" 18 : 471-473, 1998
• 2 "β-Sitosterol activates the sphingomyelin cycle and induces apoptosis in LNCaP human prostate cancer cells. Nutr Cancer 32" 32 : 8-12,
• 3 "β- Sitosterol inhibits HT-29 human colon cancer cell growth and alters membrane lipids." 16 : 2797-2804, 1996
• 4 "phase in the colonic mucosa of rats. Mech Ageing Dev" 116 : 1-14, 2000
• 5 "on the threshold of checkpoints and development. Curr Opin Cell Biol 6" 847-852,
• 6 "induces apoptosis and activates key caspases in MDA- MB-231 human breast cancer cells." 10 : 497-500,
• 7 "Zhang L. 2000. p53-independent induction of p21 reduction of cyclin B1 and G2/M arrest by the isoflavone genistein in human prostate carcinoma cells. Jpn J Cancer Res" 91 : 164-173,
• 8 "Wang XF. 1995. Functional analysis of the transforming growth factor β responsive elements in the WAF/Cip1/p21 promoter." 270 : 28623-28628,
• 9 "Vogelstain B. 1994.WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res" Cancer Res 54 : 1169-1174,
• 10 "Topical antiinflammatory activity of phytosterols isolated from Eryngium foetidum on chronic and acute inflammation models." 13 : 78-80, 1999
• 11 "The role of phytosterols and phytosterolins in immune modulation a review of the past 10 years." 4 : 471-475, 2001
• 12 "The retinoblastoma protein and cell cycle control." 81 : 323-330, 1995
• 13 "The Pezcoller lecture: cancer cell cycles revisited" 3689-3695, 2000
• 14 "Takahashi M. 1980. Protective effect of plant sterol against chemically induced colon tumors in rats. Cancer Res 40" 403-405,
• 15 "Sakai T. 1999. Histone deacetylase inhibitor activates the p21/WAF1/Cip1 gene promoter through the Sp1 sites. Ann N Y Acad Sci 886" 886 : 195-199
• 16 "Roberts JM. 1993. Cyclin-dependent regulation of G1 in mammalian fibroblasts. Science 259" 1908-1912.
• 17 "Roberts JM. 1992. Formation and activation of a cyclin E-cdk2 complex during the G1 phase of the human cell cycle. Science 257" 1689-1694,
• 18 "Regulation of cyclin D1 by calpain protease." 272 : 28479-28484, 1997
• 19 "Plant sterols and sterolins a review of their immune-modulating properties." 4 : 170-177,
• 20 "Omura S. 1987. Effects of phytosterols on anti-complementary activity. Chem Pharm Bull" 4851-4855,
• 21 "Morgan DO. 1995. Principles of CDK regulation. Nature 374" 374 : 131-134, 1995
• 22 "Miettinen TA. 1992. Effects of unsaturated and saturated dietary plant sterols on their serum contents. Clin Chim Acta 205" 97-107,
• 23 "Mareel M. 1999. The role of the E-cadherin/catenin complex in gastrointestinal cancer." 62 : 393-402, 1999
• 24 "Maller JL. 1991. Role for cyclin A in the dependence of mitosis on completion of DNA replication. Nature 354" 314-317,
• 25 "Levine JS. 1998. Necrosis and apoptosis in acute renal failure. Semin Nephrol" 18 : 505-518, 1998
• 26 "Kim KW. 2000. Induction of apoptosis by ursolic acid through activation of caspases and down-regulation of c-IAPs in human prostate epithelial cells." 17 : 565-571, 2000
• 27 "Kato JY. 1994. D-type cyclin-dependent kinase activity in mammalian cells. Mol Cell Biol 14" 2066-2076,
• 28 "Harlow E. 1994. Identification of G1 kinase activity for cdk6" 2077-2086,
• 29 "Green DR. 2001. The machinery of programmed cell death. Pharmacol Ther 92" 92 : 57-70, 2001
• 30 "Gardner JM. 1978. Sterol content of foods of plant origin. J Am Diet Assoc 73" 39-47,
• 31 "Fukuda K. 1999. Apoptosis-associated cleavage of β-catenin in human colon cancer and rat hepatoma cells. Int J Biochem Cell Biol" 31 : 519-529,
• 32 "Fink CS. 2001. Phytosterols reduce in vitro metastatic ability of MDA-MB-231 human breast cancer cells." 40 : 157-164, 2001
• 33 "Elledge SJ. 1993. The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases." 75 : 805-816,
• 34 "El-Deiry WS. 1996. Regulation of p21WAF1/CIP1 expression by p53-independent pathways. Oncogene" 12 : 1557-1564,
• 35 "E-cadherin- catenin cell-cell adhesion complex and human cancer." Pignatelli M. 2000. E-cadherin- catenin cell-cell adhesion complex and human cancer. Br J Surg 87 87 : 992-1005, 2000
• 36 "Dwyer JT. 1988. Health aspects of vegetarian diets. Am J Clin Nutr 48" 712-738,
• 37 "Bhargava KP. 1980. Anti-inflammatory and anti-pyretic activities of β-sitosterol. Planta Med 39" 157-163,
• 38 "Beach D. 1994. p21 is a component of active cell cycle kinases. Cold Spring Harb Symp Quant Biol 59" 59 : 21-29,
• 39 "Bar R. 1993. Cyclodextrins as carriers of cholesterol and fatty acids in cultivation of mycoplasmas. Appl Environ Microbiol 59" 547-551,
• 40 "Angiogenic activity of β-sitosterol in the ischaemia/reperfusion-damaged brain of Mongolian gerbil. Planta Med" 68 : 330-335, 2002
• 41 ". Research technics for the cell cycle study." 33 : -36, 2001