Neurochemical Research

, Volume 41, Issue 10, pp 2728–2751 | Cite as

Fucoxanthin Activates Apoptosis via Inhibition of PI3K/Akt/mTOR Pathway and Suppresses Invasion and Migration by Restriction of p38-MMP-2/9 Pathway in Human Glioblastoma Cells

  • Yugang Liu
  • Jian Zheng
  • Yan Zhang
  • Zhaotao Wang
  • Yang Yang
  • Miaochun Bai
  • Yiwu DaiEmail author
Original Paper


Fucoxanthin is rich in seaweed and considered as effective anti-cancer drug because of powerful antioxidant properties. The objective of this study was to investigate the role of fucoxanthin on apoptosis, invasion and migration of glioma cells. Firstly, fucoxanthin showed obvious cytotoxicity against human glioma cancer cell line U87 and U251, however, there was no inhibitory effect on normal neuron. And then, fucoxanthin induced apoptotic cell death showed by the condensation of chromatin material stained with Hoechest 33342, and reduced mitochondrial membrane potential via DIOC6(3) staining, and enhanced apoptosis by annexin V-FITC/SYTOX Green double staining on U87 and U251 cell lines. Transmission electron microscopy and western blotting were used to determine ultrastructure of U87 cell and expression of proteins related to apoptosis. A scratch wound healing assay and the expression of matrix metalloproteinases (MMPs), and a tans-well assay were used to investigate cell migration and invasion, respectively. Additionally, we uncovered upstream signaling Akt/mTOR and p38 pathways induced by incubation U87 and U251 cell lines with fucoxanthin that mediated cell apoptosis, migration and invasion by using PI3K and p38 inhibitors. Moreover, incubation of fucoxanthin obviously reduced the weight and volume of glioma mass of U87 cells in nude mice. Furthermore, we also examined the glioma mass of U87 cells by hematoxylin-eosin staining, TUNEL assay and western blot, and these outcomes in vivo consistently confirmed that above results in vitro. Taken together, these findings suggest that fucoxanthin augments apoptosis, and reduces cell proliferation, migration and invasion, and reveals a potential mechanism of fucoxanthin-mediated Akt/mTOR and p38 susspression in human glioblastoma cell line.


Fucoxanthin U87 and U251 Apoptosis Invastion Migration PI3K/Akt/mTOR P38 



This research was supported by Natural Science Foundation of China (NSFC-81271391).

Compliance with Ethical Standards

Conflict of Interest

The authors declare no conflict of interest.


  1. 1.
    Rao JS (2003) Molecular mechanisms of glioma invasiveness: the role of proteases. Nat Rev Cancer 3:489–501CrossRefPubMedGoogle Scholar
  2. 2.
    Dunbar E, Yachnis AT (2010) Glioma diagnosis: immunohistochemistry and beyond. Adv Anat Pathol 17:187–201CrossRefPubMedGoogle Scholar
  3. 3.
    Liang C, Yang L, Guo S (2015) All–retinoic acid inhibits migration, invasion and proliferation, and promotes apoptosis in glioma cells. Oncol Lett 9:2833–2838PubMedPubMedCentralGoogle Scholar
  4. 4.
    Sugawara T, Baskaran V, Tsuzuki W, Nagao A (2002) Brown algae fucoxanthin is hydrolyzed to fucoxanthinol during absorption by Caco-2 human intestinal cells and mice. J Nutr 132:946–951PubMedGoogle Scholar
  5. 5.
    Yan X, Chuda Y, Suzuki M, Nagatat T (1999) Fucoxanthin as the major antioxidant in hifikia fusiformis, a common edible seaweed. Biosci Biotechnol Biochem 63:605–607CrossRefPubMedGoogle Scholar
  6. 6.
    Hu T, Liu D, Chen Y, Wu J, Wang S (2010) Antioxidant activity of sulfated polysaccharide fractions extracted from Undaria pinnitafida in vitro. Int J Biol Macromol 46:193–198CrossRefPubMedGoogle Scholar
  7. 7.
    Sachindra NM, Sato E, Maeda H, Hosokawa M, Niwano Y, Kohno M, Miyashita K (2007) Radical scavenging and singlet oxygen quenching activity of marine carotenoid fucoxanthin and its metabolites. J Agric Food Chem 55:8516–8522CrossRefPubMedGoogle Scholar
  8. 8.
    Heo SJ, Jeon YJ (2009) Protective effect of fucoxanthin isolated from Sargassum siliquastrum on UV-B induced cell damage. J Photochem Photobiol B Biol 95:101–107CrossRefGoogle Scholar
  9. 9.
    Heo SJ, Ko SC, Kang SM, Kang HS, Kim JP, Kim SH, Lee KW, Cho MG, Jeon YJ (2008) Cytoprotective effect of fucoxanthin isolated from brown algae Sargassum siliquastrum against H2O2-induced cell damage. Eur Food Res Technol 228:145–151CrossRefGoogle Scholar
  10. 10.
    Chung TW, Choi HJ, Lee JY, Jeong HS, Kim CH, Joo M, Choic JY, Han CW, Kim SY, Choi JS, Ha KT (2013) Marine algal fucoxanthin inhibits the metastatic potential of cancer cells. Biochem Biophys Res Commun 439:580–585CrossRefPubMedGoogle Scholar
  11. 11.
    Rokkaku T, Kimura R, Ishikawa C, Yasumoto T, Senba M, Kanaya F Mori N (2013) Anticancer effects of marine carotenoids, fucoxanthin and its deacetylated product, fucoxanthinol, on osteosarcoma. Int J Oncol 43:1176–1186PubMedGoogle Scholar
  12. 12.
    Zhou T, Ye L, Bai Y, Sun A, Cox B, Liu D, Li Y, Liotta D, Snyder JP, Fu H, Huang B (2014) Autophagy and apoptosis in hepatocellular carcinoma induced by EF25-(GSH)2: a novel curcumin analog. PloS One 9:e107876CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Gao X, Deeb D, Jiang H, Liu YB, Dulchavsky SA, Gautam SC (2005) Curcumin differentially sensitizes malignant glioma cells to TRAIL/Apo2L-mediated apoptosis through activation of procaspases and release of cytochrome c from mitochondria. J Exp Ther Oncol 5:39–48PubMedGoogle Scholar
  14. 14.
    Zajc I, Hreljac I, Lah T (2006) Cathepsin L affects apoptosis of glioblastoma cells: a potential implication in the design of cancer therapeutics. Anticancer Res 26:3357–3364PubMedGoogle Scholar
  15. 15.
    Li Q, Lu XH, Cai L, Lu JL, Wu JS, Zhuge QC, Zheng WM, Su ZP (2015) Antiproliferative and apoptosis-inducing activity of schisandrin B against human glioma cells. Cancer Cell Int 15:12CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Yang JA, Li JQ, Shao LM, Yang Q, Liu BH, Wu TF, Wu P, Yi W, Chen QX (2015) Puerarin inhibits proliferation and induces apoptosis in human glioblastoma cell lines. Int J Clin Exp Med 8:10132–10142PubMedPubMedCentralGoogle Scholar
  17. 17.
    Zhang FY, Hu Y, Que ZY, Wang P, Liu YH, Wang ZH, Xue YX (2015) Shikonin inhibits the migration and invasion of human glioblastoma cells by targeting phosphorylated beta-catenin and phosphorylated PI3K/Akt: a potential mechanism for the anti-glioma efficacy of a traditional chinese herbal medicine. Int J Mol Sci 16:23823–23848CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Nie XH, Ou-Yang J, Xing Y, Li DY, Liu RE, Xu RX (2016) Calycosin inhibits migration and invasion through modulation of transforming growth factor beta-mediated mesenchymal properties in U87 and U251 cells. Drug Des Devel Ther 10:767–779PubMedPubMedCentralGoogle Scholar
  19. 19.
    Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB (1987) Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testin. Cancer Res 47:936–942PubMedGoogle Scholar
  20. 20.
    Bedner E, Li X, Gorczyca W, Melamed MR, Darzynkiewicz Z (1999) Analysis of apoptosis by laser scanning cytometry. Cytometry 35:181–195CrossRefPubMedGoogle Scholar
  21. 21.
    Dejean LM, Martinez-Caballero S, Manon S, Kinnally KW (2006) Regulation of the mitochondrial apoptosis-induced channel, MAC, by BCL-2 family proteins. BBA-Mol Basis Dis 1762:191–201CrossRefGoogle Scholar
  22. 22.
    Brumatti G, Sheridan C, Martin SJ (2008) Expression and purification of recombinant a nnexin V for the detection of membrane alterations on apoptotic cells. Methods 44:235–240CrossRefPubMedGoogle Scholar
  23. 23.
    Lecoeur H, de Oliveira-Pinto LM, Gougeon ML (2002) Multiparametric flow cytometric analysis of biochemical and functional events associated with apoptosis and oncosis using the 7-aminoactinomycin D assay. J Immunol Methods 265:81–96CrossRefPubMedGoogle Scholar
  24. 24.
    Korsmeyer SJ, Shutter JR, Veis DJ, Merry DE, Oltvai ZN (1993) Bcl-2/Bax: a rheostat that regulates an anti-oxidant pathway and cell death. Semin Cancer Biol 4:327–332PubMedGoogle Scholar
  25. 25.
    Pollack M, Phaneuf S, Dirks A, Leeuwenburgh C (2002) The role of apoptosis in the normal aging brain, skeletal muscle, and heart. Ann NY Acad Sci 959:93–107CrossRefPubMedGoogle Scholar
  26. 26.
    Sun MG, Williams J, Munoz-Pinedo C, Perkins GA, Brown JM, Ellisman MH, Green DR Frey TG (2007) Correlated three-dimensional light and electron microscopy reveals transformation of mitochondria during apoptosis. Nat Cell Boil 9:1057–1065CrossRefGoogle Scholar
  27. 27.
    Hamel W, Westphal M (2000) Growth factors in gliomas revisited. Acta Neurochir 142:113–138CrossRefPubMedGoogle Scholar
  28. 28.
    Nakano A, Tani E, Miyazaki K, Yamamoto Y, Furuyama JI (1995) Matrix metalloproteinases and tissue inhibitors of metalloproteinases in human gliomas. J Neurosurg 83:298–307CrossRefPubMedGoogle Scholar
  29. 29.
    Cox G, O’Byrne KJ (2001) Matrix metalloproteinases and cancer. Anticancer Res 21:4207–4219PubMedGoogle Scholar
  30. 30.
    Mohanam S, Gladson CL, Rao CN, Rao JS (1999) Biological significance of the expression of urokinase-type plasminogen activator receptors (uPARs) in brain tumors. Front Biosci 15:178–187CrossRefGoogle Scholar
  31. 31.
    Saiki S, Sasazawa Y, Imamichi Y, Kawajiri S, Fujimaki T, Tanida I, Kobayashi H, Sato F, Sato S, Ishikawa K, Imoto M, Hattori N (2011) Caffeine induces apoptosis by enhancement of autophagy via PI3K/Akt/mTOR/p70S6K inhibition. Autophagy 7:176–187CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Zhang L, Wang H, Zhu J, Xu J, Ding K (2004) Mollugin induces tumor cell apoptosis and autophagy via the PI3K/AKT/mTOR/p70S6K and ERK signaling pathways. Bioch Em Bioph Res Co 450:247–254CrossRefGoogle Scholar
  33. 33.
    Westermarck J, Kahari V-M (1999) Regulation of matrix metalloproteinase expression in tumour invasion. FASEB J 13:781–792PubMedGoogle Scholar
  34. 34.
    Johnson GL, Lapadat R (2002) Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 298:1911–1912CrossRefPubMedGoogle Scholar
  35. 35.
    Ahn BH, Min G, Bae YS (2006) Phospholipase D is activated and phosphorylated by casein kinase-II in human U87 astroglioma cells. Exp Mol Med 38:55–62CrossRefPubMedGoogle Scholar
  36. 36.
    Camphausen K, Purow B, Sproull M, Scott T, Ozawa T, Deen DF, Tofilon PJ (2005) Orthotopic growth of human glioma cells quantitatively and qualitatively influences radiation-induced changes in gene expression. Cancer Res 65:10389–10393CrossRefPubMedGoogle Scholar
  37. 37.
    Nakada M, Niska JA, Miyamori H, McDonough WS, Wu J, Sato H, Berens ME (2004) The phosphorylation of EphB2 receptor regulates migration and invasion of human glioma cells. Cancer Res 64:3179–3185CrossRefPubMedGoogle Scholar
  38. 38.
    Sheline GE (1986) Tumors of the brain. Springer: BerlinGoogle Scholar
  39. 39.
    Tani E, Morimura T, Kaba K, Itagaki T (1980) Preliminary study of the effects of vitamin A on antineoplastic activities of chemotherapeutic agents in glioma. Neurol Med Chir 20:665–677CrossRefGoogle Scholar
  40. 40.
    Yung WA, Lotan R, Lee P, Lotan D, Steck PA (1989) Modulation of growth and epidermal growth factor receptor activity by retinoic acid in human glioma cells. Cancer Res 49:1014–1019PubMedGoogle Scholar
  41. 41.
    Gumireddy K, Sutton LN, Phillips PC, Reddy CD (2003) All-trans-retinoic acid-induced apoptosis in human medulloblastoma: activation of caspase-3/poly(ADP-ribose) polymerase 1 pathway. Clin Cancer Res 9:4052–4059PubMedGoogle Scholar
  42. 42.
    Ying M, Wang S, Sang Y, Sun P, Lal B, Goodwin CR, Guerrero-Cazares H, Quinon s-Hinojosa A, Laterra J, Xia S (2011) Regulation of glioblastoma stem cells by retinoic acid: role for Notch pathway inhibition. Oncogene 30:3454–3467CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Liang C, Yang L, Guo S (2015) All-trans retinoic acid inhibits migration, invasion and proliferation, and promotes apoptosis in glioma cells in vitro. Oncol Lett 9:2833–2838PubMedPubMedCentralGoogle Scholar
  44. 44.
    Belloc F, Dumain P, Boisseau MR, Jalloustre C, Reiffers J, Bernard P, Lacombe F (1994) A flow cytometric method using Hoechst 33342 and propidium iodide for simultaneous cell cycle analysis and apoptosis determination in unfixed cells. Cytometry 17:59–65CrossRefPubMedGoogle Scholar
  45. 45.
    Rogakou EP, Pilch DR, Orr AH, Ivanova VS, Bonner WM (1998) DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139. J Biol Chem 273:5858–5868CrossRefPubMedGoogle Scholar
  46. 46.
    Jeong SY, Seol DW (2008) The role of mitochondria in apoptosis. BMB Rep 41:11–22CrossRefPubMedGoogle Scholar
  47. 47.
    Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312CrossRefPubMedGoogle Scholar
  48. 48.
    Saraste A, Pulkki K (2000) Morphologic and biochemical hallmarks of apoptosis. Cardiovasc Res 45:528–537CrossRefPubMedGoogle Scholar
  49. 49.
    Jiang H, Shang X, Wu H, Gautam SC, Al-Holou S, Li C, Kuo J, Zhang LJ, Chopp M (2009) Resveratrol downregulates PI3K/Akt/mTOR signaling pathways in human U251 glioma cells. J Exp Ther Oncol 8:25–33PubMedPubMedCentralGoogle Scholar
  50. 50.
    Shen J, Zheng H, Ruan J, Fang W, Li A, Tian G, Niu X, Luo S, Zhao P (2013) Autophagy inhibition induces enhanced proapoptotic effects of ZD6474 in glioblastoma. Brit. J Cancer 109:164–171CrossRefGoogle Scholar
  51. 51.
    Cantley LC (2002) The phosphoinositide 3-kinase pathway. Science 296:1655–1657CrossRefPubMedGoogle Scholar
  52. 52.
    Harris T (2003) PDK1 and PKB/Akt: ideal targets for development of new strategies to structure-based drug design. IUBMB Life 55:117–126CrossRefPubMedGoogle Scholar
  53. 53.
    Manning BD, Cantley LC (2003) United at last: the tuberous sclerosis complex gene products connect the phosphoinositide 3-kinase/Akt pathway to mammalian target of rapamycin (mTOR) signalling. Biochem Soc T 31:573–578CrossRefGoogle Scholar
  54. 54.
    Gardai SJ, Hildeman DA, Frankel SK, Whitlock BB, Frasch SC, Borregaard N, Marrack P, Bratton DL, Henson PM (2004) Phosphorylation of Bax Ser184 by Akt regulates its activity and apoptosis in neutrophils. J Biol Chem 279:21085–21095CrossRefPubMedGoogle Scholar
  55. 55.
    Song G, Ouyang G, Bao S (2005) The activation of Akt/PKB signaling pathway and cell survival. J Cell Mol Med 9:59–71CrossRefPubMedGoogle Scholar
  56. 56.
    Fata JE, Ho AV, Leco KJ, Moorehead RA, Khokha R (2000) Cellular turnover and extracellular matrix remodeling in female reproductive tissues: functions of metalloproteinases and their inhibitors. Cell Mol Life Sci 57:77–95CrossRefPubMedGoogle Scholar
  57. 57.
    Gacko M (2001) Activation mechanisms, biological role and inhibitors of metalloproteases in the extracellular matrix. Postepy Hig Med Dosw 55:303–318PubMedGoogle Scholar
  58. 58.
    Curran S, Murray GI (2000) Matrix metalloproteinases: molecular aspects of their roles in tumour invasion and metastasis. Eur J Cancer 36:1621–1630CrossRefPubMedGoogle Scholar
  59. 59.
    Deryugina EI, Bourdon MA, Luo GX, Reisfeld RA, Strongin A (1997) Matrix metalloproteinase-2 activation modulates glioma cell migration. J Cell Sci 110:2473–2482PubMedGoogle Scholar
  60. 60.
    Jung SH, Woo MS, Kim SY, Kim WK, Hyun JW, Kim EJ, Kim DH, Kim HS (2006) Ginseng saponin metabolite suppresses phorbol ester-induced matrix metalloproteinase-9 expression through inhibition of activator protein-1 and mitogen-activated protein kinase signaling pathways in human astroglioma cells. Int J Cancer 118:490–497CrossRefPubMedGoogle Scholar
  61. 61.
    Zhang Z, Lv J, Lei X, Li S, Zhang Y, Meng L, Xue R, Li Z (2014) Baicalein reduces the invasion of glioma cells via reducing the activity of p38 signaling pathway. PloS One 9:e90318CrossRefPubMedPubMedCentralGoogle Scholar
  62. 62.
    Aroui S, Najlaoui F, Chtourou Y, Meunier AC, Laajimi A, Kenani A, Fetoui H (2015) Naringin inhibits the invasion and migration of human glioblastoma cell via downregulation of MMP-2 and MMP-9 expression and inactivation of p38 signaling pathway. Tumor Biol doi: 10.1007/s13277-015-4230-4 Google Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Yugang Liu
    • 1
  • Jian Zheng
    • 2
  • Yan Zhang
    • 3
  • Zhaotao Wang
    • 1
  • Yang Yang
    • 4
  • Miaochun Bai
    • 3
  • Yiwu Dai
    • 1
    Email author
  1. 1.Affiliated Bayi Brain Hospital and Affiliated Beijing Military General Hospital of Southern Medical UniversityBeijingChina
  2. 2.Department of PathologyDezhou People HospitalJinanChina
  3. 3.Affiliated Bayi Brain Hospital, The Military General Hospital of BeijingBeijingChina
  4. 4.Dalian Medical UniversityDalianChina

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