台灣衛生福利部公告，大腸癌為102年台灣癌症十大死因第三位。癌症發展過程之生物機轉複雜，現行療法無法根除癌細胞且有副作用、復發率高，因此尋找有效替代療法是非常重要的。薑黃素（Curcumin）具有低細胞毒性、多目標阻斷路徑、抑制癌症發展的特性。文獻指出，細胞凋亡、細胞自噬、發炎因子NF-B和致癌基因Aurora-A kinase之間具有相關性。本研究在於探討薑黃素對HT-29細胞週期的影響、細胞凋亡與細胞自噬的現象及NF-B與Aurora-A kinase相關蛋白的影響。結果顯示，薑黃素增加促凋亡蛋白caspase 3、細胞自噬蛋白LC3-II表現量，代表HT-29啟動細胞凋亡與細胞自噬，抑制細胞生長。NF-B相關蛋白實驗顯示細胞中IB、phospho-IB及細胞核中NF-B蛋白量顯著下降。流式細胞儀分析觀察到薑黃素增加酸性囊泡比例，使用BAF抑制細胞自噬，降低薑黃素引起酸性囊泡比例的增加。整體而言，薑黃素影響HT-29細胞週期、引發細胞凋亡及細胞自噬、抑制HT-29細胞生長，並抑制NF-B路徑及Aurora-A相關蛋白的表現；BAF的使用可降低薑黃素引起的細胞自噬。

According to the report from the Ministry of Health and Welfare, Taiwan, 2013, human colorectal cancer is the third-leading cause of cancer death in Taiwan. Due to the complexity and high recurrent rate of cancer, as well as side effects of cancer treatment, improvement in chemotherapeutic regimens is urgently needed. Curcumin, safe in human, regulates multiple pathways to inhibit tumor progression, such as apoptosis, autophagy, inflammatory factor NF-B and oncogene Aurora-A. This study was proposed to investigate the effect of curcumin on cell cycle, apoptosis, autophagy, and protein expression and activity of NF-B and Aurora-A using human colorectal cancer HT-29 cells. The results showed that curcumin increased the expression of pro-apoptotic protein caspase 3 and autophagy protein LC3-II, suggesting that curcumin induced apoptosis and autophagy. Significant decline of IB and phospho-IBin whole cell lysates and NF-B in the nuclei were observed. Flow cytometric analysis found that the proportion of acidic vesicles increased in curcumin-treated HT-29 cells. Administration of BAF reduced curcumin-induced increment of the proportion of acidic vesicles. Overall, curcumin changed cell cycle, triggered apoptosis and autophagy, inhibited the growth of HT-29 cells, and inhibited expression of NF-B and Aurora-A. The use of BAF reduced curcumin-induced autophagy.

Aggarwal, S., Ichikawa, H., Takada, Y., Sandur, S. K., Shishodia, S., & Aggarwal, B. B. (2006).Curcumin (diferuloylmethane) down-regulates expression of cell proliferation and antiapoptotic and metastatic gene products through suppression of IkappaBalpha kinase and Akt activation. Mol Pharmacol, 69(1), 195-206.
Anand, P., Sundaram, C., Jhurani, S., Kunnumakkara, A. B., & Aggarwal, B. B. (2008). Curcumin and cancer: an "old-age" disease with an "age-old" solution. Cancer Lett, 267(1), 133-164.
Baeuerle, P. A., & Henkel, T. (1994). Function and activation of NF-kappa B in the immune system. Annu Rev Immunol, 12, 141-179.
Baldwin, A. S. (2012). Regulation of cell death and autophagy by IKK and NF-kappaB: critical mechanisms in immune function and cancer. Immunol Rev, 246(1), 327-345.
Birkenkamp, K. U., Geugien, M., Schepers, H., Westra, J., Lemmink, H. H., & Vellenga, E. (2004). Constitutive NF-kappaB DNA-binding activity in AML is frequently mediated by a Ras/PI3-K/PKB-dependent pathway. Leukemia, 18(1), 103-112.
Blakemore, L. M., Boes, C., Cordell, R., & Manson, M. M. (2013). Curcumin-induced mitotic arrest is characterized by spindle abnormalities, defects in chromosomal congression and DNA damage. Carcinogenesis, 34(2), 351-360.
Briassouli, P., Chan, F., Savage, K., Reis-Filho, J. S., & Linardopoulos, S. (2007). Aurora-A regulation of nuclear factor-kappaB signaling by phosphorylation of IkappaBalpha. Cancer Res, 67(4), 1689-1695.
Cao, A., Li, Q., Yin, P., Dong, Y., Shi, H., Wang, L., Wu, D. (2013). Curcumin induces apoptosis in human gastric carcinoma AGS cells and colon carcinoma HT-29 cells through mitochondrial dysfunction and endoplasmic reticulum stress. Apoptosis, 18(11), 1391-1402.
Dempe, J. S., Pfeiffer, E., Grimm, A. S., & Metzler, M. (2008). Metabolism of curcumin and induction of mitotic catastrophe in human cancer cells. Mol Nutr Food Res, 52(9), 1074-1081. doi: 10.1002/mnfr.200800029
Deng, L., Wang, C., Spencer, E., Yang, L., Braun, A., You, J., Chen, Z. J. (2000). Activation of the IkappaB kinase complex by TRAF6 requires a dimeric ubiquitin-conjugating enzyme complex and a unique polyubiquitin chain. Cell, 103(2), 351-361.
Dulbecco, P., & Savarino, V. (2013). Therapeutic potential of curcumin in digestive diseases. World J Gastroenterol, 19(48), 9256-9270.
Favaloro, B., Allocati, N., Graziano, V., Di Ilio, C., & De Laurenzi, V. (2012). Role of apoptosis in disease. Aging (Albany NY), 4(5), 330-349.
Fulda, S., & Debatin, K. M. (2004). Apoptosis signaling in tumor therapy. Ann N Y Acad Sci, 1028, 150-156.
Garcea, G., Berry, D. P., Jones, D. J., Singh, R., Dennison, A. R., Farmer, P. B., Gescher, A. J. (2005). Consumption of the putative chemopreventive agent curcumin by cancer patients: assessment of curcumin levels in the colorectum and their pharmacodynamic consequences. Cancer Epidemiol Biomarkers Prev, 14(1), 120-125.
Ghobrial, I. M., Witzig, T. E., & Adjei, A. A. (2005). Targeting apoptosis pathways in cancer therapy. CA Cancer J Clin, 55(3), 178-194.
Gordon, D. (1998). β-Catenin links the APC gene to MYC in colon cancer. Gastroenterology, 115(5), 1041-1042.
Grimm, S., & Baeuerle, P. A. (1993). The inducible transcription factor NF-kappa B: structure-function relationship of its protein subunits. Biochem J, 290(Pt 2), 297-308.
Grimm, S., Bauer, M. K., Baeuerle, P. A., & Schulze-Osthoff, K. (1996). Bcl-2 down-regulates the activity of transcription factor NF-kappaB induced upon apoptosis. J Cell Biol, 134(1), 13-23.
Gump, J. M., & Thorburn, A. (2011). Autophagy and apoptosis- what’s the connection? Trends Cell Biol, 21(7), 387-392.
Gupta, S., & Gollapudi, S. (2006). Molecular mechanisms of TNF-alpha-induced apoptosis in naive and memory T cell subsets. Autoimmun Rev, 5(4), 264-268.
Hatcher, H., Planalp, R., Cho, J., Torti, F. M., & Torti, S. V. (2008). Curcumin: from ancient medicine to current clinical trials. Cell Mol Life Sci, 65(11), 1631-1652.
Holy, J. M. (2002). Curcumin disrupts mitotic spindle structure and induces micronucleation in MCF-7 breast cancer cells. Mutat Res, 518(1), 71-84.
Hong, J., Bose, M., Ju, J., Ryu, J. H., Chen, X., Sang, S., Yang, C. S. (2004). Modulation of arachidonic acid metabolism by curcumin and related beta-diketone derivatives: effects on cytosolic phospholipase A(2), cyclooxygenases and 5-lipoxygenase. Carcinogenesis, 25(9), 1671-1679.
Hou, W., Han, J., Lu, C., Goldstein, L. A., & Rabinowich, H. (2010). Autophagic degradation of active caspase-8: a crosstalk mechanism between autophagy and apoptosis. Autophagy, 6(7), 891-900.
Howells, L. M., Mitra, A., & Manson, M. M. (2007). Comparison of oxaliplatin- and curcumin-mediated antiproliferative effects in colorectal cell lines. Int J Cancer, 121(1), 175-183. doi: 10.1002/ijc.22645
Huang, M. T., Lysz, T., Ferraro, T., Abidi, T. F., Laskin, J. D., & Conney, A. H. (1991). Inhibitory effects of curcumin on in vitro lipoxygenase and cyclooxygenase activities in mouse epidermis. Cancer Res, 51(3), 813-819.
Jeng, Y. M., Peng, S. Y., Lin, C. Y., & Hsu, H. C. (2004). Overexpression and amplification of Aurora-A in hepatocellular carcinoma. Clin Cancer Res, 10(6), 2065-2071.
Jin, C., & Reed, J. C. (2002). Yeast and apoptosis. Nat Rev Mol Cell Biol, 3(6), 453-459.
Jin, Z., Li, Y., Pitti, R., Lawrence, D., Pham, V. C., Lill, J. R., & Ashkenazi, A. (2009). Cullin3-based polyubiquitination and p62-dependent aggregation of caspase-8 mediate extrinsic apoptosis signaling. Cell, 137(4), 721-735.
Jurenka, J. S. (2009). Anti-inflammatory properties of curcumin, a major constituent of Curcuma longa: a review of preclinical and clinical research. Altern Med Rev, 14(2), 141-153.
Kantara, C., O'Connell, M., Sarkar, S., Moya, S., Ullrich, R., & Singh, P. (2014). Curcumin promotes autophagic survival of a subset of colon cancer stem cells, which are ablated by DCLK1-siRNA. Cancer Res, 74(9), 2487-2498.
Kashatus, D. F., Lim, K. H., Brady, D. C., Pershing, N. L., Cox, A. D., & Counter, C. M. (2011). RALA and RALBP1 regulate mitochondrial fission at mitosis. Nat Cell Biol, 13(9), 1108-1115.
Kerr, J. F. (2002, Dec 27). History of the events leading to the formulation of the apoptosis concept. Toxicology, 181-182, 471-474.
Kim, D. S., Kim, H. R., Woo, E. R., Hong, S. T., Chae, H. J., & Chae, S. W. (2005). Inhibitory effects of rosmarinic acid on adriamycin-induced apoptosis in H9c2 cardiac muscle cells by inhibiting reactive oxygen species and the activations of c-Jun N-terminal kinase and extracellular signal-regulated kinase. Biochem Pharmacol, 70(7), 1066-1078.
Kondo, Y., Kanzawa, T., Sawaya, R., & Kondo, S. (2005). The role of autophagy in cancer development and response to therapy. Nat Rev Cancer, 5(9), 726-734.
Kufer, T. A., Sillje, H. H., Korner, R., Gruss, O. J., Meraldi, P., & Nigg, E. A. (2002). Human TPX2 is required for targeting Aurora-A kinase to the spindle. J Cell Biol, 158(4), 617-623.
Lev-Ari, S., Maimon, Y., Strier, L., Kazanov, D., & Arber, N. (2006). Down-regulation of prostaglandin E2 by curcumin is correlated with inhibition of cell growth and induction of apoptosis in human colon carcinoma cell lines. J Soc Integr Oncol, 4(1), 21-26.
Levine, B., Mizushima, N., & Virgin, H. W. (2011). Autophagy in immunity and inflammation. Nature, 469(7330), 323-335.
Li, L., & Ma, B. B. (2014). Colorectal cancer in Chinese patients: current and emerging treatment options. Onco Targets Ther, 7, 1817-1828.
Littlepage, L. E., Wu, H., Andresson, T., Deanehan, J. K., Amundadottir, L. T., & Ruderman, J. V. (2002). Identification of phosphorylated residues that affect the activity of the mitotic kinase Aurora-A. Proc Natl Acad Sci U S A, 99(24), 15440-15445.
Liu, H. S., Ke, C. S., Cheng, H. C., Huang, C. Y., & Su, C. L. (2011). Curcumin-induced mitotic spindle defect and cell cycle arrest in human bladder cancer cells occurs partly through inhibition of aurora A
Lu, J. J., Cai, Y. J., & Ding, J. (2011). Curcumin induces DNA damage and caffeine-insensitive cell cycle arrest in colorectal carcinoma HCT116 cells. Mol Cell Biochem, 354(1-2), 247-252.
Marin, Y. E., Wall, B. A., Wang, S., Namkoong, J., Martino, J. J., Suh, J., Ryu, J. H. (2007). Curcumin downregulates the constitutive activity of NF-kappaB and induces apoptosis in novel mouse melanoma cells. Melanoma Res, 17(5), 274-283.
Mathew, R., Karp, C. M., Beaudoin, B., Vuong, N., Chen, G., Chen, H. Y., . . . White, E. (2009). Autophagy suppresses tumorigenesis through elimination of p62. Cell, 137(6), 1062-1075.
Mohandas, K. M., & Desai, D. C. (1999). Epidemiology of digestive tract cancers in India. V. Large and small bowel. Indian J Gastroenterol, 18(3), 118-121.
Moscat, J., & Diaz-Meco, M. T. (2009). p62 at the Crossroads of Autophagy, Apoptosis, and Cancer. Cell, 137(6), 1001-1004.
Mosieniak, G., Adamowicz, M., Alster, O., Jaskowiak, H., Szczepankiewicz, A. A., Wilczynski, G. M., . . . Sikora, E. (2012). Curcumin induces permanent growth arrest of human colon cancer cells: link between senescence and autophagy. Mech Ageing Dev, 133(6), 444-455.
Nikonova, A. S., Astsaturov, I., Serebriiskii, I. G., Dunbrack, R. L., Jr., & Golemis, E. A. (2013). Aurora A kinase (AURKA) in normal and pathological cell division. Cell Mol Life Sci, 70(4), 661-687.
Pan, M. H., Chang, W. L., Lin-Shiau, S. Y., Ho, C. T., & Lin, J. K. (2001). Induction of apoptosis by garcinol and curcumin through cytochrome c release and activation of caspases in human leukemia HL-60 cells. J Agric Food Chem, 49(3), 1464-1474.
Paraskevi Briassouli, F. C., Kay Savage, et al. (2007). Aurora-A Regulation of Nuclear Factor-κB Signaling by Phosphorylation of IκBα.
Pattingre, S., Tassa, A., Qu, X., Garuti, R., Liang, X. H., Mizushima, N., Levine, B. (2005). Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell, 122(6), 927-939.
Plummer, S. M., Holloway, K. A., Manson, M. M., Munks, R. J., Kaptein, A., Farrow, S., & Howells, L. (1999). Inhibition of cyclo-oxygenase 2 expression in colon cells by the chemopreventive agent curcumin involves inhibition of NF-kappaB activation via the NIK/IKK signalling complex. Oncogene, 18(44), 6013-6020.
Prajapati, S., Tu, Z., Yamamoto, Y., & Gaynor, R. B. (2006). IKKalpha regulates the mitotic phase of the cell cycle by modulating Aurora A phosphorylation. Cell Cycle, 5(20), 2371-2380.
Rao, C. V., Simi, B., & Reddy, B. S. (1993). Inhibition by dietary curcumin of azoxymethane-induced ornithine decarboxylase, tyrosine protein kinase, arachidonic acid metabolism and aberrant crypt foci formation in the rat colon. Carcinogenesis, 14(11), 2219-2225.
Sandur, S. K., Deorukhkar, A., Pandey, M. K., Pabon, A. M., Shentu, S., Guha, S., Krishnan, S. (2009). Curcumin modulates the radiosensitivity of colorectal cancer cells by suppressing constitutive and inducible NF-kappaB activity. Int J Radiat Oncol Biol Phys, 75(2), 534-542.
Savarino, E., Zentilin, P., Mastracci, L., Dulbecco, P., Marabotto, E., Gemignani, L., Savarino, V. (2013). Microscopic esophagitis distinguishes patients with non-erosive reflux disease from those with functional heartburn. J Gastroenterol, 48(4), 473-482.
Shakibaei, M., Mobasheri, A., Lueders, C., Busch, F., Shayan, P., & Goel, A. (2013). Curcumin enhances the effect of chemotherapy against colorectal cancer cells by inhibition of NF-kappaB and Src protein kinase signaling pathways. PLoS One, 8(2), e57218.
Shishodia, S., Amin, H. M., Lai, R., & Aggarwal, B. B. (2005). Curcumin (diferuloylmethane) inhibits constitutive NF-kappaB activation, induces G1/S arrest, suppresses proliferation, and induces apoptosis in mantle cell lymphoma. Biochem Pharmacol, 70(5), 700-713.
Soncini, C., Carpinelli, P., Gianellini, L., Fancelli, D., Vianello, P., Rusconi, L., Moll, J. (2006). PHA-680632, a novel Aurora kinase inhibitor with potent antitumoral activity. Clin Cancer Res, 12(13), 4080-4089.
Su, C. L., Huang, L. L. H., Huang, L. M., Lee, J. C., Lin, C. N., & Won, S. J. (2006). Caspase-8 acts as a key upstream executor of mitochondria during justicidin A-induced apoptosis in human hepatoma cells. Febs Letters, 580(13), 3185-3191.
Sun, C., Chan, F., Briassouli, P., & Linardopoulos, S. (2007). Aurora kinase inhibition downregulates NF-kappaB and sensitises tumour cells to chemotherapeutic agents. Biochem Biophys Res Commun, 352(1), 220-225.
Thorburn, J., Horita, H., Redzic, J., Hansen, K., Frankel, A. E., & Thorburn, A. (2009). Autophagy regulates selective HMGB1 release in tumor cells that are destined to die. Cell Death Differ, 16(1), 175-183.
Van Erk, M. J., Teuling, E., Staal, Y. C., Huybers, S., Van Bladeren, P. J., Aarts, J. M., & Van Ommen, B. (2004). Time- and dose-dependent effects of curcumin on gene expression in human colon cancer cells. J Carcinog, 3(1), 8.
Viatour, P., Merville, M. P., Bours, V., & Chariot, A. (2005). Phosphorylation of NF-kappaB and IkappaB proteins: implications in cancer and inflammation. Trends Biochem Sci, 30(1), 43-52.
Walter, A. O., Seghezzi, W., Korver, W., Sheung, J., & Lees, E. (2000). The mitotic serine/threonine kinase Aurora2/AIK is regulated by phosphorylation and degradation. Oncogene, 19(42), 4906-4916.
Wang, J. B., Qi, L. L., Zheng, S. D., & Wu, T. X. (2009). Curcumin induces apoptosis through the mitochondria-mediated apoptotic pathway in HT-29 cells. J Zhejiang Univ Sci B, 10(2), 93-102.
Wang, R. C., Wei, Y., An, Z., Zou, Z., Xiao, G., Bhagat, G., Levine, B. (2012). Akt-mediated regulation of autophagy and tumorigenesis through Beclin 1 phosphorylation. Science, 338(6109), 956-959.
Wang, Y. T., Liu, H. S., & Su, C. L. (2014). Curcumin-enhanced chemosensitivity of FDA-approved platinum (II)-based anti-cancer drugs involves downregulation of nuclear endonuclease G and NF-kappaB as well as induction of apoptosis and G2/M arrest. Int J Food Sci Nutr, 65(3), 368-374.
Warner, S. L., Bearss, D. J., Han, H., & Von Hoff, D. D. (2003). Targeting Aurora-2 kinase in cancer. Mol Cancer Ther, 2(6), 589-595.
Won, S. J., Yen, C. H., Liu, H. S., Wu, S. Y., Lan, S. H., Jiang-Shieh, Y. F., Su, C. L. (2014). Justicidin A-Induced Autophagy Flux Enhances Apoptosis of Human Colorectal Cancer Cells via Class III PI3K and Atg5 Pathway. J Cell Physiol.
Yue, Z., Jin, S., Yang, C., Levine, A. J., & Heintz, N. (2003). Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor. Proc Natl Acad Sci U S A, 100(25), 15077-15082.