题名

荷葉黃酮萃取物抑制肥胖因子促進人類乳癌細胞的轉移之作用

并列篇名

The flavonoid-rich extract from Nelumbo Nucifera leave inhibits obesity factor induced breast cancer metastasis

DOI

10.6834/CSMU.2013.00208

作者

游孟勳

关键词

荷葉黃酮萃取物 ； 脂肪細胞因子 ； 乳癌細胞株(MCF-7) ； Nelumbo Nucifera leave extract (NLE) ； adipocyte conditioned medium(CM) ； Breast cancer cell line(MCF-7)

期刊名称

中山醫學大學生化暨生物科技研究所學位論文

卷期/出版年月

2013年

学位类别

碩士
导师

王朝鐘
内容语文

繁體中文
中文摘要

台灣婦女乳腺癌是最常見的癌症，乳腺癌死亡率佔所有癌症死亡 人數的 28.4％，並且與肥胖有關。先前的研究利用荷葉萃取物(NLE) 已被証實可以有效地減少脂肪肝、高脂血症、動脈粥樣硬化和抗氧化 能力。本研究中利用脂肪細胞分泌肥胖因子而得的培養液(CM)誘導 人類乳癌細胞遷移的模式，並以 Boyden chamber 驗證，從結果得知當給予 NLE 時可以減緩在 CM 誘導狀態下乳腺癌細胞的遷移作用，隨著 NLE 劑量增加抑制的效果更明顯，且我們亦使用 ECIS 及傷口癒合試驗加以確認，最後所得到的結果皆一致。為了探討乳癌細胞遷移的分子機制，利用二維膠體電泳分析，從結果中得知在 CM 誘導的狀態下，細胞 Hsp90、StarD10、Peroxiredoxin 6 蛋白表現較高，推測NLE或許是透過這些蛋白進而影響遷移的能力；再者，我們使用西方墨點法分析人類乳癌細胞株(MCF-7)在 CM 誘導的狀態下，並且處理NLE觀察其蛋白的表現，從結果得知當處理NLE時會降低 JNK、PI3K/AKT、StarD10 和 Peroxiredoxin 6 等蛋白表現量，進而達到抑制人類乳癌細胞的遷移作用。綜上所述，本研究的結果表明未來對於治療轉移性乳癌，NLE 具有發展成為抑制癌症轉移過程中的天然藥物之潛力。
英文摘要

Breast cancer is the most common cancer in women in the Taiwan. Then breast cancer deaths accounted for 28.4% of all the cancer deaths, and have been associated with obesity. Previous studies, has been confirmed Nelumbo nucifera leave extract (NLE) can effectively reduce the fatty liver, atherosclerosis, hyperlipidemia and anti-oxidative properties. Thus, we investigated the relationship between obesity and breast cancer, and evaluated the inhibitory effect of NLE on the adipocytes-induced breast cancer progression. Adipocyte secreted conditioned medium (CM) noticeably induced breast cancer migration. NLE diminished the CM-induced migration number of breast cancer cell by boyden chamber assay. A direct correlation was found between adipocyte and breast cancer cell migration. Similarly, we used of ECIS assay, wound healing assay and transwell assay identical results were found. To investigate the molecular mechanisms of breast cancer cell migration using two-dimensional gel electrophoresis. We found CM increased the proteins level of the Hsp90, StarD10, Peroxiredoxin 6. The results suggest that CM stimulates migration of breast cancer cells, perhaps through the interaction of these proteins. Under the CM treatment, phosphorylated JNK, PI3K/Akt, StarD10 and Peroxiredoxin 6 levels of MCF-7 cells treated with NLE can significantly inhibited the protein expression were measured by Western blot analysis. These data indicate adipocytes provide a major component of the microenvironment for rapid tumor growth and migration, and this increase was inhibited significantly by NLE treatment. In summary, the study indicated that the future for the treatment of metastatic breast cancer, NLE inhibited cancer metastasis has developed into a process of natural drugs.
主题分类 醫藥衛生 > 基礎醫學
醫學院 > 生化暨生物科技研究所
参考文献
  1. 1　行政院衛生署. 民國101年十大死因統計年報. 2013.
    連結：
  2. 2　Li Puma, G., Puddu, V., Tsang, H. K., Gora, A. & Toepfer, B., Photocatalytic oxidation of multicomponent mixtures of estrogens (estrone (E1), 17β-estradiol (E2), 17α-ethynylestradiol (EE2) and estriol (E3)) under UVA and UVC radiation: Photon absorption, quantum yields and rate constants independent of photon absorption. Applied Catalysis B: Environmental2010, 99, 388-397.
    連結：
  3. 3　Kastelic, J., Bergfelt, D. & Ginther, O., Relationship between ultrasonic assessment of the corpus luteum and plasma progesterone concentration in heifers. Theriogenology 1990, 33, 1269-1278.
    連結：
  4. 4　Martinez, V. & Azzopardi, J., Invasive lobular carcinoma of the breast: incidence and variants. Histopathology 1979, 3, 467-488.
    連結：
  5. Revision of the American Joint Committee on Cancer staging system for
    連結：
  6. 6　Livraghi T, Goldberg SN, Solbiati L, Meloni F, Ierace T, Gazelle GS., Percutaneous Radio-frequency Ablation of Liver Metastases from Breast Cancer: Initial Experience in 24 Patients1. Radiology 2001, 220, 145-149.
    連結：
  7. 8　行政院衛生署國民健康局. Body Mass Index,BMI標準. 2013.
    連結：
  8. make a difference in the ability of obese individuals to lose body weight. International Journal of Obesity 2012, 37, 759-764.
    連結：
  9. 10　Anzman, S., Rollins, B. & Birch, L., Parental influence on children's earlyeating environments and obesity risk: implications for prevention. International Journal of Obesity 2010, 34, 1116-1124.
    連結：
  10. 11　Lajunen, H.-R., Kaprio, J., Rose, R. J., Pulkkinen, L. & Silventoinen, K.,Genetic and environmental influences on BMI from late childhood to adolescence are modified by parental education. Obesity 2011, 20, 583-589.
    連結：
  11. 13　Lin, E. & Tsai, S.-J., Novel Diagnostics RD for Public Health and Personalized Medicine in Taiwan: Current State, Challenges and Opportunities. Current Pharmacogenomics and Personalized Medicine 2012, 10, 239-246.
    連結：
  12. 14　Gouda HN, Sagoo GS, Harding AH, Yates J, Sandhu MS, Higgins JP., The association between the peroxisome proliferator-activated receptor-γ2 (PPARG2) Pro12Ala gene variant and type 2 diabetes mellitus: a HuGE review and meta-analysis. American journal of epidemiology 2010, 171, 645-655.
    連結：
  13. 17　Bartke, A., Impact of reduced insulin‐like growth factor‐1/insulin signaling on aging in mammals: novel findings. Aging cell 2008, 7, 285-290.
    連結：
  14. 19　Bays HE, González-Campoy JM, Bray GA, Kitabchi AE, Bergman DA, Schorr AB, Rodbard HW, Henry RR., Pathogenic potential of adipose tissue and metabolic consequences of adipocyte hypertrophy and increased visceral adiposity. Expert Review of Cardiovascular Therapy 2008, 6, 343-368.
    連結：
  15. 20　Ogden, C. L., Carroll, M. D., Kit, B. K. & Flegal, K. M., Prevalence of obesity and trends in body mass index among US children and adolescents, 1999-2010. JAMA: the journal of the American Medical Association 2012, 307, 483-490.
    連結：
  16. 21 Campion, J., Checinski, K., Nurse, J. & McNeill, A., Smoking by people with mental illness and benefits of smoke-free mental health services. Advances in Psychiatric Treatment 2008, 14, 217-228.
    連結：
  17. 22　Hursting, S. D., Smith, S. M., Lashinger, L. M., Harvey, A. E. & Perkins, S. N., Calories and carcinogenesis: lessons learned from 30 years of calorie restriction research. Carcinogenesis 2010, 31, 83-89.
    連結：
  18. 23　Folkman, J., Angiogenesis in cancer, vascular, rheumatoid and other disease.
    連結：
  19. Nature Medicine 1995, 1, 27-31.
    連結：
  20. 24　Fidler, I. J., The pathogenesis of cancer metastasis: the seed and soil hypothesis revisited. Nature Reviews Cancer 2003, 3, 453-458.
    連結：
  21. 25　Cano A, Pérez-Moreno MA, Rodrigo I, Locascio A, Blanco MJ, del Barrio MG, Portillo F, Nieto MA., The transcription factor snail controls epithelial–mesenchymal transitions by repressing E-cadherin expression. Nature cell biology 2000, 2, 76-83.
    連結：
  22. 26 Lee, J. M., Dedhar, S., Kalluri, R. & Thompson, E. W., The epithelial–mesenchymal transition: new insights in signaling, development, and disease. The Journal of cell biology 2006, 172, 973-981.
    連結：
  23. 27　Ewertz, M., Gillanders, S., Meyer, L. & Zedeler, K., Survival of breast cancer patients in relation to factors which affect the risk of developing breast cancer. International journal of cancer 1991, 49, 526-530.
    連結：
  24. 29 Millis, K. K., Maas, W. E., Cory, D. G. & Singer, S., Gradient, high‐resolution, magic‐angle spinning nuclear magnetic resonance spectroscopy of human adipocyte tissue. Magnetic resonance in medicine 1997, 38, 399-403.
    連結：
  25. 30 Kucerova, L., Altanerova, V., Matuskova, M., Tyciakova, S. & Altaner, C. Adipose tissue–derived human mesenchymal stem cells mediated prodrug cancer gene therapy. Cancer research 2007, 67, 6304-6313.
    連結：
  26. 31 Garofalo C, Koda M, Cascio S, Sulkowska M, Kanczuga-Koda L, Golaszewska J, Russo A, Sulkowski S, Surmacz E., Increased expression of leptin and the leptin receptor as a marker of breast cancer progression: possible role of obesity-related stimuli. Clinical Cancer Research 2006, 12, 1447-1453.
    連結：
  27. 32 Brown, M. D., Hart, C. A., Gazi, E., Bagley, S. & Clarke, N. W., Promotion of prostatic metastatic migration towards human bone marrow stoma by Omega 6 and its inhibition by Omega 3 PUFAs. British journal of cancer 2006, 94, 842-853.
    連結：
  28. 36 Kashiwada Y, Aoshima A, Ikeshiro Y, Chen YP, Furukawa H, Itoigawa M, Fujioka T, Mihashi K, Cosentino LM, Morris-Natschke SL, Lee KH., Anti-HIV benzylisoquinoline alkaloids and flavonoids from the leaves of Nelumbo nucifera, and structure–activity correlations with related alkaloids. Bioorganic & medicinal chemistry 2005, 13, 443-448.
    連結：
  29. 37 石田均司, 海野孝之, 辻邦朗 & 小菅卓夫., Studies on the Antihemorrhagic Substances in Herbs Classified as Hemostatics in Chinese Medicine. VIII.: On the Antihemorrhagic Principle in Nelumbins Receptaculum. Chemical & pharmaceutical bulletin 1988, 36, 4585-4587.
    連結：
  30. 38 Rai, S., Wahile, A., Mukherjee, K., Saha, B. P. & Mukherjee, P. K., Antioxidant activity of Nelumbo nucifera (sacred lotus) seeds. Journal of ethnopharmacology 2006, 104, 322-327.
    連結：
  31. 39 Ling, Z. Q., Xie, B. J. & Yang, E. L., Isolation, Characterization, and Determination of Antioxidative Activity of Oligomeric Procyanidins from the Seedpod of Nelumbo nucifera Gaertn. Journal of Agricultural and Food Chemistry 2005, 53, 2441-2445.
    連結：
  32. 40 Ono, Y., Hattori, E., Fukaya, Y., Imai, S. & Ohizumi, Y., Anti-obesity effect of Nelumbo nucifera leaves extract in mice and rats. Journal of ethnopharmacology 2006, 106, 238-244.
    連結：
  33. 41 Wu CH, Yang MY, Chan KC, Chung PJ, Ou TT, Wang CJ., Improvement in high-fat diet-induced obesity and body fat accumulation by a Nelumbo nucifera leaf flavonoid-rich extract in mice. Journal of Agricultural and Food Chemistry 2010, 58, 7075-7081.
    連結：
  34. 42 Du H, You JS, Zhao X, Park JY, Kim SH, Chang KJ., Antiobesity and hypolipidemic effects of lotus leaf hot water extract with taurine supplementation in rats fed a high fat diet. Journal of biomedical science 2010, 17, 42-47.
    連結：
  35. 43　洪云佳. 荷葉黃酮萃取物抑制第二型糖尿病副病變之作用 The flavonoid-rich extract from Nelumbo Nucifera leave inhibited complications of type II diabetes mellitus. 中山醫學大學生化暨生物科技研究所學位論文 2012, 1-167.
    連結：
  36. 44 Lin MC, Kao SH, Chung PJ, Chan KC, Yang MY, Wang CJ., Improvement for high fat diet-induced hepatic injuries and oxidative stress by flavonoid-enriched extract from Nelumbo nucifera leaf. Journal of Agricultural and Food Chemistry 2009, 57, 5925-5932.
    連結：
  37. 45 李季螢. 荷葉乙醇萃取物對動脈粥狀硬化早期發展之影響. 國立台灣大學食品科技研究所碩士論文 2004.
    連結：
  38. 52 Yang, M.-Y., Chang, Y.-C., Chan, K.-C., Lee, Y.-J. & Wang, C.-J. Flavonoid-enriched extracts from Nelumbo nucifera leaves inhibits proliferation of breast cancer in vitro and in vivo. European Journal of Integrative Medicine 2011, 3, 153-163.
    連結：
  39. 53 Wu CH, Yang MY, Chan KC, Chung PJ, Ou TT, Wang CJ., Improvement in high-fat diet-induced obesity and body fat accumulation by a Nelumbo nucifera leaf flavonoid-rich extract in mice. Journal of agricultural and food chemistry 2010, 58, 7075-7081.
    連結：
  40. 54 Du H, You JS, Zhao X, Park JY, Kim SH, Chang KJ., Antiobesity and hypolipidemic effects of lotus leaf hot water extract with taurine supplementation in rats fed a high fat diet. Journal of biomedical science 2010, 17, 42-47.
    連結：
  41. 58 Rolli, M., Fransvea, E., Pilch, J., Saven, A. & Felding-Habermann, B., Activated integrin αvβ3 cooperates with metalloproteinase MMP-9 in regulating migration of metastatic breast cancer cells. Proceedings of the National Academy of Sciences 2003, 100, 9482-9487.
    連結：
  42. 59 Hazan, R. B., Phillips, G. R., Qiao, R. F., Norton, L. & Aaronson, S. A., Exogenous expression of N-cadherin in breast cancer cells induces cell migration, invasion, and metastasis. The Journal of cell biology 2000, 148, 779-790.
    連結：
  43. 60 del Barco Barrantes, I. & Nebreda, A., Roles of p38 MAPKs in invasion and metastasis. Biochemical Society Transactions 2012, 40, 79.
    連結：
  44. 61 Huo Y, Qiu WY, Pan Q, Yao YF, Xing K, Lou MF., Reactive oxygen species (ROS) are essential mediators in epidermal growth factor (EGF)-stimulated corneal epithelial cell proliferation, adhesion, migration, and wound healing. Experimental eye research 2009, 89, 876-886.
    連結：
  45. 62 Welf, E. S., Ahmed, S., Johnson, H. E., Melvin, A. T. & Haugh, J. M., Migrating fibroblasts reorient directionality by a metastable, PI3K-dependent mechanism. The Journal of cell biology 2012, 197, 105-114.
    連結：
  46. 64 Ho HH, Hsu LS, Chan KC, Chen HM, Wu CH, Wang CJ., Extract from the leaf of nucifera reduced the development of atherosclerosis via inhibition of vascular smooth muscle cell proliferation and migration. Food and Chemical Toxicology 2010, 48, 159-168.
    連結：
  47. 65 Guerrero J, Tobar N, Cáceres M, Espinoza L, Escobar P, Dotor J, Smith PC, Martínez J., Soluble factors derived from tumor mammary cell lines induce a stromal mammary adipose reversion in human and mice adipose cells. Possible role of TGF-β1 and TNF-α. Breast cancer research and treatment 2010, 119, 497-508.
    連結：
  48. 66 E Griner, S., J Wang, K., P Joshi, J. & Nahta, R., Mechanisms of Adipocytokine-Mediated Trastuzumab Resistance in HER2-Positive Breast Cancer Cell Lines. Current Pharmacogenomics and Personalized Medicine (Formerly Current Pharmacogenomics) 2013, 11, 31-41.
    連結：
  49. 67 Takaoka M, Suzuki H, Shioda S, Sekikawa K, Saito Y, Nagai R, Sata M., Endovascular injury induces rapid phenotypic changes in perivascular adipose tissue. Arteriosclerosis, thrombosis, and vascular biology 2010, 30, 1576-1582.
    連結：
  50. 68 Ho HH, Hsu LS, Chan KC, Chen HM, Wu CH, Wang CJ., Extract from the leaf of nucifera reduced the development of atherosclerosis via inhibition of vascular smooth muscle cell proliferation and migration. Food and Chemical Toxicology 2010, 48, 159-168.
    連結：
  51. 69 Creydt, V. P., Sacca, P. A., Tesone, A. J., Vidal, L. & Calvo, J. C., Adipocyte differentiation influences the proliferation and migration of normal and tumoral breast epithelial cells. Molecular Medicine Reports 2010, 3, 433-439.
    連結：
  52. 70 Chen, C.-Y., Abell, A. M., Moon, Y. S. & Kim, K.-H., An advanced glycation end product (AGE)-receptor for AGEs (RAGE) axis restores adipogenic potential of senescent preadipocytes through modulation of p53 protein function. The Journal of biological chemistry 2013, 288, 10949.
    連結：
  53. 71 Olayioye MA, Vehring S, Müller P, Herrmann A, Schiller J, Thiele C, Lindeman GJ, Visvader JE, Pomorski T., StarD10, a START domain protein overexpressed in breast cancer, functions as a phospholipid transfer protein. Journal of Biological Chemistry 2005, 280, 27436-27442.
    連結：
  54. 72 Olayioye MA, Hoffmann P, Pomorski T, Armes J, Simpson RJ, Kemp BE, Lindeman GJ, Visvader JE., The phosphoprotein StarD10 is overexpressed in breast cancer and cooperates with ErbB receptors in cellular transformation. Cancer research 2004, 64, 3538-3544.
    連結：
  55. 73 Lee SB, Ho JN, Yoon SH, Kang GY, Hwang SG, Um HD., Peroxiredoxin 6 promotes lung cancer cell invasion by inducing urokinase-type plasminogen activator via p38 kinase, phosphoinositide 3-kinase, and Akt. Molecules and cells 2009, 28, 583-588.
    連結：
  56. 5　Singletary SE, Allred C, Ashley P, Bassett LW, Berry D, Bland KI, Borgen PI, Clark G, Edge SB, Hayes DF, Hughes LL, Hutter RV, Morrow M, Page DL, Recht A,Theriault RL, Thor A, Weaver DL, Wieand HS, Greene FL.,
  57. breast cancer. Journal of Clinical Oncology 2002, 20, 3628-3636.
  58. 7　盧崇亮. 轉移性乳腺癌的治療進展. 中國現代普通外科進展 2006, 9,
  59. 10-13.
  60. 9　Tremblay, A., Royer, M., Chaput, J. & Doucet, E., Adaptive thermogenesis can
  61. 12　Moleres, A., Martinez, J. & Marti, A., Genetics of Obesity. Current Obesity Reports 2013, 1-9.
  62. 15　Liu Y, Zhai C, Cao P, Zheng Y, Zhou S, Chi J., FABP2 gene polymorphism and metabolic disorder of lipids in the middle aged and aged population. Journal of hygiene research 2011, 40, 461.
  63. 16　Heisler LK, Jobst EE, Sutton GM, Zhou L, Borok E, Thornton-Jones Z, Liu HY, Zigman JM, Balthasar N, Kishi T, Lee CE, Aschkenasi CJ, Zhang CY, Yu J, Boss O, Mountjoy KG, Clifton PG, Lowell BB, Friedman JM, Horvath T, Butler AA, Elmquist JK, Cowley MA., Serotonin reciprocally regulates melanocortin neurons to modulate food intake. Neuron 2006, 51, 239-249.
  64. 18　Kathiresan S, Melander O, Guiducci C, Surti A, Burtt NP, Rieder MJ, Cooper GM, Roos C, Voight BF, Havulinna AS, Wahlstrand B, Hedner T, Corella D, Tai ES,Ordovas JM, Berglund G, Vartiainen E, Jousilahti P, Hedblad B, Taskinen MR, Newton-Cheh C, Salomaa V, Peltonen L, Groop L, Altshuler DM, Orho-Melander M., Six new loci associated with blood low-density lipoprotein cholesterol, high-density lipoprotein cholesterol or triglycerides in humans. Nature genetics 2008, 40, 189-197.
  65. 28 Ewertz M, Jensen MB, Gunnarsdóttir KÁ, Højris I, Jakobsen EH, Nielsen D, Stenbygaard LE, Tange UB, Cold S., Effect of obesity on prognosis after early-stage breast cancer. Journal of clinical oncology 2011, 29, 25-31.
  66. 33 Nieman KM, Kenny HA, Penicka CV, Ladanyi A, Buell-Gutbrod R, Zillhardt MR, Romero IL, Carey MS, Mills GB, Hotamisligil GS, Yamada SD, Peter ME, Gwin K,Lengyel E., Adipocytes promote ovarian cancer metastasis and provide energy for rapid tumor growth. Nature medicine 2011, 17, 1498-1503.
  67. 34 Yang, Y., Yen, S. & Lin, C., Illustrated guide to aquatic plants of Taiwan. Council of Agriculture, The Executive Yuan of Taiwan 2001.
  68. 35 陳錦木. 蓮荷產業發展與蓮花栽培技術改進. 水生植物多樣性開發與利用研討會專刊 2009.
  69. 46 陶波，帥景賢，吳鳳蓮. 荷葉水煎劑對高脂血症大鼠血脂及血液流變學的影響. 中醫藥學報 2000, 6, 55-56.
  70. 47 杜力軍, 孫., 李敏, 金文, 徐麗珍. 荷葉大豆及其合劑調脂活性部位的研究. 中草藥 2000, 7, 526-528.
  71. 48 楊藏雄, 陳., 吳素卿, 陳國棟. 臺灣產蓮Nelumbo nucifera GAERTEN 之生物鹼研究. 臺灣藥學雜誌 15, 76-79.
  72. 49 余以剛，陳海光，曾慶孝. 荷葉水提物清除自由基的ESR研究. 中草藥 2001, 32, 693-69.
  73. 50 涂予文. 薏仁、山楂、荷葉與陳皮水萃物對倉鼠脂質代謝的影響. 國立台灣大學食品科技研究所碩士論文 2002.
  74. 51 朱淯維. 荷葉乙醇萃取物抗氧化性之探討. 國立台灣大學食品科技研究所碩士論文 2003.
  75. 55　Ortega FJ, Moreno-Navarrete JM, Mayas D, García-Santos E, Gómez-Serrano M, Rodriguez-Hermosa JI, Ruiz B, Ricart W, Tinahones FJ, Frühbeck G, Peral B,Fernández-Real JM., Breast Cancer 1 (BrCa1) May Be behind Decreased Lipogenesis in Adipose Tissue from Obese Subjects. PloS one 2012, 7, 33233.
  76. 56 Lopes-Virella, M. F., Stone, P., Ellis, S. & Colwell, J., Cholesterol determination in high-density lipoproteins separated by three different methods. Clinical Chemistry 1977, 23, 882-884.
  77. 57 Middelbeek J, Kuipers AJ, Henneman L, Visser D, Eidhof I, van Horssen R, Wieringa B, Canisius SV, Zwart W, Wessels LF, Sweep FC, Bult P, Span PN, van Leeuwen FN, Jalink K., TRPM7 is required for breast tumor cell metastasis. Cancer research 2012, 72, 4250-4261.
  78. 63 陳海光, 曾慶孝., 荷葉功能成分的提取及其對自由基清除作用的研究. 食品及發酵工業 2002, 27, 34-38.
print cancel