臺灣博碩士論文加值系統

在台灣傳統民俗療法中，土半夏被認為具散瘀、止血、消腫、解毒及抗癌功能，但這些療效至今尚未被科學化地驗証。本研究主要探討土半夏之抗癌活性及其作用機轉。將新鮮土半夏之葉片以酒精萃取，並依序以正己烷、乙酸乙酯及甲醇萃取分層分離。由抗氧化活性檢測結果顯示，乙酸乙酯層萃取物具有良好的 DPPH 自由基清除能力，其 EC50 值為 89.59 ± 0.06 μg/ml。其次，以 MTT 分析方法測試各萃取物對不同癌細胞株之毒殺效果，包括乳癌細胞 MCF-7、MDA-MB-231、肺癌細胞 A549、H661、攝護腺癌細胞 LNCaP、DU145、肝癌細胞 Hep G2、Hep 3B 及腦瘤細胞 GBM，其中以酒精萃取所得之粗萃物對 LNCaP 細胞毒殺效果最佳，其 IC50 值為 124.5 ± 0.2 μg/ml，且隨劑量及處理時間之增加，抑制癌細胞生長的作用愈明顯。當投藥濃度為 200 μg/ml 時，處理時間為 24 hr 時，細胞存活率為 47.0 ± 0.3 %；由 GC-MS 分析結果顯示，phytol 為土半夏酒精粗萃物內之主成份之一，其可能提供了土半夏酒精粗萃物對於 LNCaP 細胞之毒殺效力。將處理過土半夏酒精粗萃物之 LNCaP 細胞以流式細胞儀觀察顯示，此粗萃物會導致 LNCaP 細胞生長週期滯留在 G2/M 期，且此一作用之程度與添加劑量呈正相關效應。在 caspase 3 活性方面，土半夏酒精粗萃物並不能誘導 LNCaP 細胞進行 caspase 3 之活化。最後，利用西方墨點法進一步證實，土半夏酒精粗萃物是藉由 cyclin D3 蛋白質表現之下降來導致細胞週期滯留在 G2/M 期。由以上結果顯示，土半夏酒精粗萃物具有抑制人類攝護腺癌細胞生長之功效，且其機轉亦被探討，未來或許可用於臨床上治療攝護腺癌之輔助治療藥物。

In Taiwan, Typhonium divaricatum (L.) Decne is a folk medicine herb endowing with reducing ecchymosis, hemostasis, detumescence, detoxication, anticancer, and so on. However, these bioactivities have not yet been examined scientifically. This study investigated the anticancer activities of T. divaricatum and its mechanisms. Fresh leaves of T. divaricatum were extracted with ethanol and then fractionated by n-hexane, ethyl acetate (EA) and methanol. In antioxidant activity assay, EA fraction showed a good DPPH radical scavenging activity (EC50 = 89.59 ± 0.06 μg/ml). The cytotoxic effects of these extracts on various cancer cell lines, including MCF-7, MDA-MB-231 (breast cancer), A549, H661 (lung cancer), LNCaP, DU145 (prostate cancer), HepG2, Hep3B (hepatoma), and GBM (brain tumor), were determined by MTT assay. The ethanol extract exhibited a highest cytotoxicity on LNCaP cells (IC50 = 124.5 ± 0.2 μg/ml). The ethanol extract induced cytotoxic effect on LNCaP cells in dose- and time-dependent manners. When LNCaP cells were treated with 200 μg/ml for 24 hr, the cell viability was 47.0 ± 0.3%. According to GC-MS analysis, phytol is one of the major components in T. divaricatum ethanol extract. It may provide the vital cytotoxicity of T. divaricatum ethanol extract on LNCaP cells. The flow cytometric analysis indicated that the LNCaP cells were arrested at G2/M phase in cells treated with ethanol extract. The ratio of the cells arrested at G2/M phase was dose-dependent with the ethanol extract treatment. Moreover, the caspase-3 assay revealed that the ethanol extract could not induce the activation of caspase-3 in LNCaP cells. The G2/M phase arrest inducing by ethanol extract maybe mediated through the reduction of cyclin D3 level. These results suggested that the ethanol extract of T. divaricatum possessed the cytotoxicity on human prostate cancer cells. In addition, the mechanism had also been investigated. In the future, the herb might be used as an adjuvant medicine on the clinical treatment of prostate cancer.

中文摘要………………………………………………………………I
英文摘…………………………………………………………………II
致謝……………………………………………………………………III
目錄……………………………………………………………………IV
圖目錄…………………………………………………………………VII
表目錄…………………………………………………………………VIII
附圖表目…………………………………………………………… .IX
1. 緒論 …………………………………………………………...1
1.1 中草藥保健食品之保健開發………………………………...3
1.2 土半夏之保健功能………………………………………….. 5
1.3 土半夏相關研究……………………………………………… 6
1.4 研究目的……………………………………………………… 7
2. 自由基…………………………………………………………… 9
2.1 自由基定義…………………………………………………… 9
2.2 自由基的產生………………………………………………… 9
2.3 過多自由基造成的傷害……………………………………… 10
3. 細胞增殖與調控……… …………………………………… ….12
3.1 細胞死亡之途徑……………………………………………… 12
3.2 細胞週期…………………… …………………………………14
3.3 調控細胞週期之蛋白和激酶………………………………… 15
3.4 Cyclins 週期蛋白家族.………………………………………16
3.5 Cyclin-dependent kinases (CDKs) 週期蛋白依賴激酶家
族……………………………………………………………… 18
3.6 Cyclin-dependent kinase inhibitors (CDKI) 週期蛋白依賴
激酶抑制劑…………………………………………………….18
3.7 細胞週期檢查點(Cell cycle checkpoints) ………………19
3.8細胞週期與各週期蛋白和激酶間之關連性……………………20
3.9 細胞凋亡時的生化變化與特徵.………………………………22
3.10 細胞凋亡時之相關因素.…………………………………… 24
4. 材料與方法.………………………………………………………26
4.1土半夏之萃取……………………………………………………26
4.2管柱色層分析……………………………………………………26
4.3薄膜色層分析(TLC)…………………………………………… 26
4.4 抗氧化活性分析……………………………………………… 27
4.4.1 DPPH 自由基清除活性之測定.………………………… 27
4.4.2 清除超氧陰離子能力測定…………………………………28
4.5 抗氧化有效成分之總量分析………………………………… 30
4.5.1總酚(Total polyphenols)測定法.……………………… 30
4.5.2類黃酮(Flavonoids)含量測定…………………………… 30
4.6 細胞培養條件及方法………………………………………… 31
4.7 細胞型態變化觀察…………………………………………… 34
4.8 細胞存活率之測定－MTT assay………………………………34
4.9 細胞週期之研究……………………………………………… 35
4.10 Caspases 酵素活性測定…………………………………… 36
4.11 蛋白質之定性及定量分析……………………………………38
4.12 西方墨點法(Western Blot) .………………………………39
4.13 GC-Mass analysis……………………………………… … 44
4.14 數據分析………………………………………………………44
4.15 實驗流程………………………………………………………45
5. 實驗結果………………………………………………………… 47
5.1 土半夏之萃取純化與各分層萃取物之極性分析…………… 47
5.2 抗氧化活性及成分之定量分析……………………………… 50
5.3 土半夏萃取物對不同細胞之毒殺作用及形態的改變……… 53
5.4 細胞凋亡之測定……………………………………………… 61
5.5 GC-Mass 質譜儀分析………………………………………… 70
6. 結論……………………………………………………………… 72
7. 參考文獻.…………………………………………………………73

Abraham, A., Macnicol, M. F. Growth arrest lines and recurrent patellar dislocation: a new sign. Knee. 2001, 8, 163-165.

Agami, R., Bernards, R. Distinct initiation and maintenance mechanisms cooperate to induce G1 cell cycle arrest in response to DNA damage. Cell. 2000, 102, 55-66.

Alt, F. W., Pinho, R., Zimmerman, K., Legouy, E., Hatton, K., Ferrier, P., Tesfaye, A., Yancopoulos, G., Nisen, P. The human myc gene family. Cold Spring Harb Symp. 1986, 51, 931-941.

Ames. B. N. DNA damage from micronutrient deficiencies is likely to be a major cause of cancer. Fund & Mol Mech of M. 2001, 475, 7-20.

Andersen, G., Busso, D., Poterszman, A., Hwang, J. R., Wurtz, J. M., Ripp, R.,Thierry, J. C., Egly, J. M., Moras, D. The structure of cyclin H: common mode of kinase activation and specific features. EMBO J. 1997, 16, 958-967.

Baldin, V., Lukas, J., Marcote, M. J., Pagano, M., Draetta, G. Cyclin D1 is a nuclear protein required for cell cycle progression in G1. Gene Dev. 1993, 7, 812-821.

Bartek, J., Bartkova, J., Lukas, J. The retinoblastoma protein pathway in cell cycle control and cancer. Exp cell Res. 1997, 237, 1-6.

Baserga, R., Wiebel, F. The cell cycle of mammalian cells. Int Rev of Exp Pathol.1969, 7, 1-30.

Bhuvaneswari, V., Nagini, S. Lycopene: a review of its potential as an anticancer agent. Curr Med Chem Anticancer Age. 2005, 5, 627-635.

Bodnarchuk, I. A. Analysis of the role of DNA repair, regulation of cell cycle and apoptosis in the radiation-induced adaptive response of mammalian cell. Radiat Biol Radioecol. 2003, 43, 19-28.
Boone, C. W., Kelloff, G. J., Malone, W. E. Identification of candidate cancer chemopreventive agents and their evaluation in animal models and human clinical trials. Cancer Res. 1990, 50, 2-9.

Bortner, D. M., Rosenberg, M. P. Overexpression of cyclin A in the mammary glands of transgenic mice results in the induction of nuclear abnormalities and increased apoptosis. Cell Growth Differ. 1995, 6, 1579-1589.

Briganti, S., Picarod, M. Antioxidant activity, lipid peroxidation and skin disease. J Eur Acad Dermatol Venereol. 2003, 17, 663-669.

Brooks, G., Poolman, R. A., Li, J. M. Arresting developments in the cardiac myocyte cell cycle: Role of cyclin-dependents kinase inhibitors. Cardiovasc Res. 1998, 39, 301-311.

Brooks, R., Fantes, P., Hunt, T., Wheatley, D. The Cell Cycle. Cambridge: The Company of Biologists Ltd, 1989.

Calixto, J. B., Santos, A. R., Filbo, V. C., Yunes, R. A. A review of the plants of the genus Phyllanthus: their chemistry, pharmacology, and therapeutic potential. Med Res Rev. 1998, 18 (4), 225-258.

Chang, C. K., Yu, H. J., Chan, K. W., Lai, M. K. Secular trend and age-period cohort analysis of prostate cancer mortality in Taiwan. J Urol. 1997, 158, 1845-1848.

Charles, J. S., James, M. R. CDK inhibitors: positive and negative regulators of G1-phase progression. Gene Dev. 1999, 13, 1501-1512.

Childs, N. M., Poryzees, G. H. Foods that help prevent disease: consumer attitudes and public policy implications. Brit Food J. 1998, 9, 419-426.

Choo, C. Y., Chan, K. L., Sam, T. W., Hitotsuyanagi, Y., Takeya, K. The cytotoxicity and chemical constituents of the hexane fraction of Typhonium flagelliforme (Araceace). Phytother Res. 2001, 77(1), 129-131.

Donaldson, K. L, Goolsby, G., Kiener, P. A., Wahl, A. F. Activation of p34cdc2 coincident with Taxol-induced apoptosis. Cell Growth Differ. 1994, 5, 1041-1050.
Courjal, F., Louason, G., Speiser, P., Katsaros, D., Zeillinger, R., Theillet, C. Cyclin gene amplification and overexpression in breast and ovarian cancers: evidence for
the selection of cyclin D1 in breast and cyclin E in ovarian tumors. Int J Cancer. 1996, 69, 247-253.

Draetta, G., Eckstein, J. Cdc25 protein phosphatases in cell proliferation. Bioch et Bioph Acta. 1997, 1332, 53-63.

Driscoll, J. S., Hazard, G. F., Wood, H. B., Goldin, A. Structure-antitumor activity relationships among quinone derivatives. Cancer Chemother Rep. 1974, 2, 1-362.

Enari, M., Talanian, R. V., Wong, W. W. and Nagata, S. Sequential activation of ICE-like and CPP32-like protease during Fas-mediated apoptosis. Nature. 1996, 380, 723-726.

Esser, P., Heimann, K., Abts, H., Fontana, A., Weller, M. CD95 (Fas/APO-1) antibody-mediated apoptosis of human retinal pigment epithelial cells. Biochem Biophy Res Commu. 1995, 213, 1026-1034.

Fahey, J. W., Zhang, Y., Talalay, P. Broccoli sprouts: An exceptionally rich source of inducers of enzyme that protect against chemical carcinogens. Proc Nad Acad Sci.
1997, 94, 10367 – 10372.

Fenech, M., Ferguson, L. R. Vitamins/minerals and genomic stability in humans.Fund & Mol Mech of M. 2001, 475,1-6.

Fil’chenkov, A. A. Apoptosis modulators. Biomed Khim. 2003, 49, 333-359.

Fukushima, S., Takada, N., Hori, T., Wanibuchi, H. Cancer prevention by organosulfur compounds from garlic and onion. J Cell Biochem Suppl. 1997, 27,
100-105.

Giridharan, P. Novel substituted methylenedioxy lignan suppresses proliferation of cancer cells by inhibiting telomerase and activation of c-myc and caspases leading
to apoptosis. Br J Cancer, London. 2002, 87(1), 98-105.

Graves, P. R., Yu, L., Schwarz, J. K., Gales, J., Sausville, E. A., O''Connor, P. M., Piwnica-Worms, H. The Chk1 protein kinase and the Cdc25C regulatory pathways are targets of the anticancer agent UCN-01. J Biol Chem. 2000, 275, 5600-5605.

Greenwald. P. Clinical trials of breast and prostate cancer prevention. J Nutr. 2001, 131, 176-178.

Gross, N., Balmas, K., Beretta Brognara, C., Tschopp, J. Expression of Fas (APO-1/CD95) and Fas ligand (FasL) in human neuroblastoma. Med Pediatr Oncol. 2001, 36, 111-114.

Harold, E. M., Fred, R., Leonard, M., Aruna, P., Kanter, M. Antioxidant content of whole grain breakfast cereals, fruits and vegetables. J Am College Nutr. 2000, 19, 312S-319S.
Halliwell, B., Gutteridge, J. M. C. Free Radicals in Biology and Medicine, 2nd ed. Clarendon, Oxford. 1989, 49-73.

Halliwell, B., Gutteridge, J. M., Cross, C. E. Free radicals, antioxidants, and human disease: Where are we now? The J Lab Clin Med. 1992, 119, 598-620.

Hanna, D. E., Rethinaswamy, A., Glover, C. V. Casein kinase II is required for cell cycle progression during G1 and G2/M in Saccharomyces cerevisiae. J Biol Chem. 1995, 270, 25905-25914.

Hermeking, H., Lengauer, C., Polyak, K., He, T. C., Zhang, L., Thiagalingam, S., Kinzler, K. W. Vogelstein B. 14-3-3 sigma is a p53-regulated inhibitor of G2/M progression. Mol Cell. 1997, 1, 3-11.

Hinchcliffe, E. H., Li, C., Thompson, E. A., Maller, J. L., Sluder, G. Requirement of Cdk2-cyclin E activity for repeated centrosome reproduction in Xenopus egg extracts. Science. 1999, 283, 851-854.

Hunter, T., Pines, J. Cyclin D and CDK inhibitors come of age Cell. Cyclins and cancer II. 1994, 79, 573-582.

Hsu, H. F., Houng, J. Y., Chang, C. L. Wu, C. C., Chang, F. R., Wu, Y. C. Antioxidant activity, cytotoxicity, and DNA information of Glossogyne tenuifolia. J Agric Food Chem. 2005, 53, 6117-6125.

Hutchison, C., Glover, D. M. Cell Cycle Control. New York: Oxford University Press. 1995.

Ibrado, A. M., Huang, Y., Fang, G., Liu, L., Bhalla, K. Overexpression of Bcl-2 or Bcl-xL inhibits Ara-C-induced CPP32/Yama protease activity and apoptosis of human acute myelogenous leukemia HL- 60 cells. Cancer Res. 1996, 56,
4743-4748.

Ishizaki, Y. Physiological functions of programmed cell death. Seikagaku. 1998, 70, 365-370.

Kan, S. F., Huang, W. J., Lin, L. C., Wang, P. S. Inhibitory effects of evodiamine on the growth of human prostate cancer cell line LNCaP. Int J Cancer. 2004, 10, 110(5), 641-651.

Kasai, H. Oxidative DNA lesions and mutagenesis. J Enviro Mutagen Res. 1996, 18, 181-189.

Kawabe, T., Suganuma, M., Ando, T., Kimura, M., Hori, H., Okamoto, T. Cdc25C interacts with PCNA at G2/M transition. Oncogene. 2002, 21, 1717-1726.

Kawanishi, S., Hiraku, Y., Oikawa, S. Mechanism of guanine-specific DNA damage by oxidative stress and its role in carcinogenesis and aging. Mutat Res. 2001, 488, 65-76.

Kelloff, G. J., Boone, C. W., Crowell, J. A., Nayfield, S. G., Hawk, E., Malone,W. F., Steele, V. E., Lubet, R. A., Sigman, C. C. Risk biomarkers and current strategies for cancer chemoprevention. J Cell Biochem Suppl. 1996, 25, 1-14.

Kingston, D. G. Recent advances in the chemistry of taxol. J Nat prod. 2000, 63, 726-734.

Koepp, D. M., Harper, J. W., Elledge, S. J. How the cyclin became a cyclin: regulated proteolysis in the cell cycle. Cell. 1999, 97, 431-434.

Koff, A., Cross, F., Fisher, A., Schumacher, J., Leguellec, K., Philippe, M., Roberts, J. M. Human cyclin E, a new cyclin that interacts with two members of the CDC2 gene family. Cell. 1991, 66, 1217-1728.

Kostyuk, V. A., Potapovich, A. I. Antiradical and chelating effects in flavonoid protection against silica-induced cell injury. Arch. Biochem. Biophys. 1998, 355, 43-48.

Leonhard, M., Gruber, P., Chott, P. A. Mutations in apoptosis genes: a pathogenetic factor for human disease. Mutat Res. 2001, 488, 211-231.

Li, H., Zhu, H., Xu, C. J., Yuan, J. Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. Cell .1998, 94, 491-501.

Li, J. M., Brooks, G. Cell cycle regulatory molecules (cyclins, cyclin-dependent kinases and cyclin-dependent kinase inhibitors) and the cardiovascular system. Eur Heart J. 1999, 20, 406-420.

Ling, Y. H., Consoli, U., Tornos, C., Andreeff, M., Perez-Soler, R. Accumulation of cyclin B1, activation of cyclin B1-dependent kinase and induction of programmed cell death in human epidermoid carcinoma KB cells treated with taxol. Int J Cancer. 1998, 75, 925-932.

Ling, Y. H., Jiang, J. D., Holland, J. F., Perez-Soler, R. Arsenic trioxide produces polymerization of microtubules and mitotic arrest before apoptosis in human tumor cell lines. Mol Pharmacol. 2002, 62, 529-538.

Li, X. K., Motwani, M., Tong, W., Bornmann, W., Schwartz, G. K. Huanglian, A chinese herbal extract, inhibits cell growth by suppressing the expression of cyclin B1 and inhibiting CDC2 kinase activity in human cancer cells. Mol Pharmacol. 2000, 58, 1287-1293.

Lundberg, A. S., Weinberg, R. A. Control of the cell cycle and apoptosis. Eur J Cancer. 1999, 35, 1886-1894.

Martinez, A. M., Afshar, M., Martin, F., Cavadore, J. C., Labbé, J. C., Dorée, M. Dual phosphorylation of the T-loop in cdk7: its role in controlling cyclin H
binding and CAK activity. The EMBO J. 1997, 16, 343-354.

McDonnell, T. J., Beham, A., Sarkiss, M., Andersen, M. M., Lo, P. Importance of the Bcl-2 family in cell death regulation. Experientia. 1996, 52, 1008-1017.

Meikrantz,W., Schlegel, R. Suppression of apoptosis by dominant negative mutants of cyclin-dependent protein kinases. J Biol Chem. 1996, 271, 10205-10209.

Meraldi, P., Lukas, J., Fry, A. M., Bartek, J., Nigg, E. A. Centrosome duplication in mammalian somatic cells requires E2F and Cdk2-cyclin A. Nat Cell Biol. 1999, 1, 88-93.

Michalides, R. J. Cell cycle regulators: mechanisms and their role in aetiology, prognosis, and treatment of cancer. J Clin Pathol. 1999, 52, 555-568.

Miller, D. K. The role of the caspase family of cysteine proteases in apoptosis, Semin Immunol. 1997, 9, 35-49.

Morgan, D. O. Principles of CDK regulation. Nature. 1995, 374, 131-134.

Musacchio, A., Hardwick, K. G. The spindle checkpoint: structural insights into dynamic signalling. Nat Rev Mol Cell Bio. 2002, 3, 731-741.

Napolitano, A., Cascone, A., Graziani, G., Ferracane, R., Scalfi, L., Di Vaio, C.,Ritieni, A., Fogliano, V. Influence of variety and storage on the polyphenol composition of apple flesh. J Agric Food Chem. 2004, 52, 6526-6531.

Nicholson, D. W., Ali, A., Thornberry, N. A., Vaillancourt, J. P., Ding, C. K., Callant, M., Gareau, Y., Griffin, R. R., Labelle, M., Lazebnik, Y. A., Munday, N. A., Raju, S. M., Smulson, M. E., Yamin, T. T., Yu, V. L. and Miller, D. K. Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature 1995, 376, 37-43.

Ohtani, K., DeGregori, J., Nevins, J. R. Regulation of the cyclin E gene by transcription factor E2F1. Proceedings of the National Academy of Sciences of the United States of America. 1995, 92, 12146-12150.
Otsuki, Y., Li, Z., Shibata, M. A. Apoptotic detection method－from morphology to gene. Prog. Histochem. Cyto. 2003, 38, 275-339.

Panaro, N. J., Popesu, N. C., Thorgeirsson, U. P. Flavone acetic acid induced a G2/M cell cycle arrest in mammary carcinoma cells. Brit J Cancer. 1999, 80, 905-1911.

Paulovich, A. G., Margulies, R. U., Garvik, B. M., Hartwell, L. H. RAD9, RAD17,and RAD24 are required for S phase regulation in Saccharomyces cerevisiae in response to DNA damage. Genetics. 1997, 145, 45-62.

Pines, J. Cyclins and cyclin-dependent kinases: a biochemical view. Biochem J. 1995,308, 697-711.

Pines, J. Cyclin-dependent kinase inhibitors: the age of crystals. Biochem Biophys Acta. 1997, 1332, 39-42.

Polyak, K., Lee, M. H., Erdjument-Bromage, H., Koff, A., Roberts, J. M., Tempst, P., Massague, J. Cloning of p27Kip1, a cyclin-dependent kinase inhibitor and a potential mediator of extracellular antimitogenic signals. Cell. 1994, 78, 59-66.

Reed, J. C. Double identity for proteins of the Bcl-2 family. Nature. 1997, 387, 773-776

Reed, J. C. Mechanisms of apoptosis avoidance in cancer. Curr Opin Oncol. 1999, 11, 68-75.

Reed, J. C. Regulation of apoptosis by bcl-2 family proteins and its role in cancer and chemoresistance. Curr Opin Oncol. 1995, 7, 541-546.

Reed, S. I., Bailly, E., Dulic, V., Hengst, L., Resnitzky, D., Slingerland, J. G1 control in mammalian cells. J Cell Sci - Supplement. 1994, 18, 69-73.

Robertson, M. J., Manley, T. J., Pichert, G., Cameron, C., Cochran, K. J., Levine, H., Ritz, J. Functional consequences of APO-1/Fas (CD95) antigen expression by
normal and neoplastic hematopoietic cells. Leukemia Lymphoma. 1995, 17, 51-61.

Rochelle, L. G., Fischer, B. M., Adler, K. B. Concurrent production of reactive oxygen and nitrogen species by airway epithelial cells in vitro. Free Radic Bio Med. 1998, 24, 863-868.
Rodenas, J., Mitjavila, M. T. Stimultaneous generation of nitric oxide and superoxide by inflammatory cells in rats. Free Radic Bio Med. 1995, 18, 869-875.

Romani, A., Coinu, R., Carta, S., Pinelli, P., Galardi, C., Vincieri, F. F., Franconi, F.Evaluation of antioxidant effect of different extracts of Myrtus communis L. Free
Radical Res. 2004, 38, 97-103.

Saint-Cricq de Gaulejac N, Vivas N, de Freitas V, Burgeois G: The influence of various phenolic compounds on scavenging activity assessed by an enzymatic method. J Sci Food Agric. 1999, 79, 1081-1090.

Sanchez, A., Alvarez, A. M., Benito, M., Fabregat, I. Cycloheximide prevents apoptosis, reactive oxygen species production, and glutathione depletion induced by transforming growth factor beta in fetal rat hepatocytes in primary culture. Hepatology. 1997, 26, 935-943.

Shah, M. A., Schwartz, G. K. Cell cycle-mediated drug resistance: an emerging concept in cancer therapy. Clin Cancer Res. 2001, 7, 2168-2181.

Sherr, C. J., Roberts, J. M. Inhibitors of mammalian G1 cyclin－dependent kinases. Gene Dev. 1995, 9, 1149-1163.

Sherr, C. J., Roberts, J. M. CDK inhibitors: positive and negative regulators of G1-phase progression. Gene Dev. 1999, 13, 1501-1512.

Shivapurkar, N., Reddy, J., Chaudhary, P. M.,Gazdar, A. F. Apoptosis and lung cancer: a review. J Cell Biochem. 2003, 88, 885-898.

Shyu, Y.S., Hwang, L.S. Antioxidant activity of the crude extract of lignan glycosides from unroasted Burma black sesame meal. Food Res Int. 2002, 35, 357-365.

Smith, M. L., Fornace, Jr. A. J. Mammalian DNA damage-induciblegenes associated with growth arrest and apoptosis. Mutat Res. 1996, 340, 109-124.

Stewart, Z. A., Pietenpol, J. A. p53 Signaling and cell cycle checkpoints. Chem Res Toxicol. 2001, 14, 243-263.
Sutherland, C., Campbell, D. G., Cohen, P. Identification of insulin-stimulated protein kinase-1 as the rabbit equivalent of rskmo-2. Identification of two threonines
phosphorylated during activation by mitogen-activated protein kinase. European J Biophy. 1993, 212, 581-588.

Swamy, S. M. K., Tan, B. K. H. Cytotoxic and mmunopotentiating effects of ethanolic extract of Nigella sativa L. seeds. J Ethnopharmacol. 2000, 70, 1-7.

Talanian, R. V., Quinlan, C., Trautz, S., Hackett, M. C., Mankovich, J. A., Banach, D., Ghayur, T., Brady, K. D., Wing, W. W. Substrate specificities of caspase family proteases. J Biol Chem. 1997, 272, 9677-9682.

Tompson, C. B. Apoptosis in the pathogenesis and treatment of disease. Sci. 1995, 267, 1456-1462.

Trabera, M. G., Atkinson, J. Vitamin E, antioxidant and nothing more. Free Radical Bio Med. 2007, 43, 4–15.

Tyson, J. J., Novak, B., Odell, G. M., Chen, K., Tron, C. D. Chemical kinetic theory: understanding cell-cycle regulation. Trends Biochem Sci. 1996, 12, 89-95.

Van Cruchten, S., Van den, Broeck. W. Morphological and biochemical aspects of apoptosis, oncosis and necrosis. Anat Histol Embryol. 2002, 31, 214-223.

Van Poppel, G., Van den Berg, H. Vitamins and cancer. Cancer Lett. 1997, 114, 195-202

Vecchione, A., Ishii, H., Baldassarre, G., Bassi, P., Trapasso, F., Alder, H., Pagano, F., Gomella, L. G. Croce C. M. Baffa, R. FEZ1/LZTS1 is down-regulated in high-grade bladder cancer, and its restoration suppresses umorigenicity in transitional cell carcinoma cells. Am J Pathol. 2002, 160, 1345-1352.

Wang, J. F., Komarov, P., Groot, H. Luminol-hemiluminescence in rat macrophages and granulocytes : the role of NO, O2•-/H2O2 and HOCl. Arch Biochem Biophys. 1993, 304, 189-196.

Weisburger, J. H. Evaluation of the evidence on the role of tomato products in disease prevention. Pro Soc Exp Biol Med. 1998, 218, 104-143.
William, C. A., Harborne, J. B., Mayo, S. J. Anthocyanin pigments and leaf flavonoids in the family Araceae. Phytochemistry. 1981, 20, 217-234.

Wimmel, A., Lucibello, F. C., Sewing, A., Adolph, S., Muller, R. Inducible acceleration of G1 progression through tetracycline-regulated expression of human cyclin E. Oncogene. 1994, 9, 995-997.

Wolfe, K., Wu, X., Liu, R. H. Antioxidant activity of apple peels. J Agr Food Chem. 2003, 51, 609-614.

Wu, M. J., Wang, L., Ding, H. Y., Wang, C. Y., Yen, J. H. Glossogyne tenuifolia acts inhibut inflammatory mediator production in a macrophage cell line by downregulating LPS-induced NF-kappa B. J Biomed Sci. 2004, 11, 186-199.

Yanez, J., Vicente, V., Alcaraz, M., Castillo, J., Benavente-Garcia, O., Canteras, M., Teruel, J. A. Cytotoxicity and antiproliferative activities of several phenolic compounds against three melanocytes cell lines: relationship between structure and activity. Nutr Cancer. 2004, 49, 191-199.
Yang, K., Liu, S., Zhang, E. Zhongcaoyao, 1985, 16(3), 138.

You, B. Y., Wang, Y. H., Kuo, M. L. Role of reactive oxygen species in cupric 8-quinolinnoxide-induced genotoxic effect. Mutat Res. 2001, 491, 45-56.

Yu, J., Liu, H., Lei, J., Tan, W., Hu, X., Zou, G. Antitumor Activity of Chloroform Fraction of Scutellaria barbata and its Active Constituents. Phytother. Res. 2007, DOI: 10.1002/ptr.2062.

Zheng, A., Mantymaa, P., Saily, M., Savolainen, E., Vahakangas, K., Koistinen P. p53 pathway in apoptosis induced by all-trans-retinoic acid in acute myeloblastic
leukaemia cells. Acta haematol-basel. 2000, 103, 135-43.

Zi, X., Agarwal, R. Silibinin decreases prostate-specific antigen with cell growth inhibition via G1 arrest, leading to differentiation of prostate carcinoma cells: implications for prostate cancer intervention. Proc Natl Acad Sci U S A. 1999, 96, 7490-7495.

Zi, X., Grasso, A. W., Kung, H. J., Agarwal, R. A flavonoid antioxidant, silymarin, inhibits activation of erbB1 signaling and induces cyclin-dependent kinase inhibitors, G1 arrest, and anticarcinogenic effects in human prostate carcinoma DU145 cells. Cancer Res. 1998, 58, 1920-1929.

Zornig, M., Hueber, A. O., Evan, G. Apoptosis regulators and their role in tumorigensis. Bioche et Biophy Acta. 2001, 1551, F1-F37.

Zou, Y., Lu, Y., Wei, D. Antioxidant activity of a flavonoid rich extract of Hypericum perforatum L. in vitro. J Agr Food Chem. 2004, 52, 5032-5039.

林天送.自由基與健康. 科學月刊. 1994, 25, 515-521.

林天送.人體自由基知多少? 健康世界雜誌. 1994, 228, 20-24.

佐佐木舜一.台灣民間藥用植物誌. 1924.

呂鋒洲.蕃茄與防癌. 宏欣文化. 2000

薛仲娟.土半夏 Typhonium divaricatum (L.) Decne 之成份研究，國立清華大學化學研究所碩士論文. 1988.

甘淑芬.吳茱萸鹼對人類前列腺癌細胞生長之抑制作用，國立陽明大學醫學院生理學研究所博士論文. 2003

黃寬裕.苦蘵引起人類乳癌細胞G2/M期停滯和細胞凋亡，中國醫藥大學醫學研究所. 2004