DOI QR코드

DOI QR Code

Beta-asarone Induces LoVo Colon Cancer Cell Apoptosis by Up-regulation of Caspases through a Mitochondrial Pathway in vitro and in vivo

  • Zou, Xi (Department of Oncology, The Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Traditional Chinese Medicine) ;
  • Liu, Shen-Lin (Department of Oncology, The Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Traditional Chinese Medicine) ;
  • Zhou, Jin-Yong (Central Laboratory, The Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Traditional Chinese Medicine) ;
  • Wu, Jian (Central Laboratory, The Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Traditional Chinese Medicine) ;
  • Ling, Bo-Fan (Department of Oncology, The Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Traditional Chinese Medicine) ;
  • Wang, Rui-Ping (Department of Oncology, The Affiliated Hospital of Nanjing University of Chinese Medicine, Jiangsu Province Hospital of Traditional Chinese Medicine)
  • Published : 2012.10.31

Abstract

Beta-asarone is one of the main bioactive constituents in traditional Chinese medicine Acorus calamu. Previous studies have shown that it has antifungal and anthelmintic activities. However, little is known about its anticancer effects. This study aimed to determine inhibitory effects on LoVo colon cancer cell proliferation and to clarify the underlying mechanisms in vitro and in vivo. Dose-response and time-course anti-proliferation effects were examined by MTT assay. Our results demonstrated that LoVo cell viability showed dose- and time-dependence on ${\beta}$-asarone. We further assessed anti-proliferation effects as ${\beta}$-asarone-induced apoptosis by annexin V-fluorescein isothiocyanate/propidium iodide assay usinga flow cytometer and observed characteristic nuclear fragmentation and chromatin condensation of apoptosis by microscopy. Moreover, we found the apoptosis to be induced through the mitochondrial/caspase pathway by decreasing mitochondrial membrane potential (MMP) and reducing the Bcl-2-to-Bax ratio, in addition to activating the caspase-9 and caspase-3 cascades. Additionally, the apoptosis could be inhibited by a pan-caspase inhibitor, carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK). When nude mice bearing LoVo tumor xenografts were treated with ${\beta}$-asarone, tumor volumes were reduced and terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) assays of excised tissue also demonstrated apoptotic changes. Taken together, these findings for the first time provide evidence that ${\beta}$-asarone can suppress the growth of colon cancer and the induced apoptosis is possibly mediated through mitochondria/caspase pathways.

References

  1. Bernales S, McDonald KL and Walter P (2006). Autophagy counterbalances endoplasmic reticulum expansion during the unfolded protein response. PLoS Biol, 4, e423. https://doi.org/10.1371/journal.pbio.0040423
  2. Cho SG, Choi, EJ (2002). Apoptotic signaling pathways: caspases and stress-activated protein kinases. J Biochem Mol Biol, 35, 24-7. https://doi.org/10.5483/BMBRep.2002.35.1.024
  3. Cummings J, Ward TH, Ranson M, et al (2004). Apoptosis pathway-targeted drugs--from the bench to the clinic. Biochim Biophys Acta, 1705, 53-66.
  4. Degterev A, Yuan J (2008). Expansion and evolution of cell death programmes. Nat Rev Mol Cell Biol, 9, 378-90. https://doi.org/10.1038/nrm2393
  5. Denault JB, Boatright K (2004). Apoptosis in Biochemistry and Structural Biology. 3-8 February 2004, Keystone, CO, USA. IDrugs, 7, 315-7.
  6. Ewings KE, Wiggins CM, Cook SJ (2007). Bim and the prosurvival Bcl-2 proteins: opposites attract, ERK repels. Cell Cycle, 6, 2236-40. https://doi.org/10.4161/cc.6.18.4728
  7. Fang YQ, Fang RM Fang, GL, et al (2008). Effects of beta-asarone on expression of c-fos in kindling epilepsy rat brain. Zhongguo Zhong Yao Za Zhi, 33, 534-6.
  8. Fu SY, Fang RM, Fang GL, et al (2008). Effects of beta-asarone on expression of FOS and GAD65 in cortex of epileptic rat induced by penicillin. Zhong Yao Cai, 31, 79-81.
  9. Ghavami S, Eshragi M, Ande SR, et al (2010). S100A8/A9 induces autophagy and apoptosis via ROS-mediated crosstalk between mitochondria and lysosomes that involves BNIP3. Cell Res, 20, 314-31. https://doi.org/10.1038/cr.2009.129
  10. Ghavami S, Kerkhoff C, Chazin WJ, et al (2008). S100A8/9 induces cell death via a novel, RAGE-independent pathway that involves selective release of Smac/DIABLO and Omi/HtrA2. Biochim Biophys Acta, 1783, 297-311. https://doi.org/10.1016/j.bbamcr.2007.10.015
  11. Jemal A, Bray F, Center MM, et al (2011). Global cancer statistics. CA Cancer J Clin, 61, 69-90. https://doi.org/10.3322/caac.20107
  12. Kim J, Seo SM, Lee SG, et al (2008). Nematicidal activity of plant essential oils and components from coriander (Coriandrum sativum), Oriental sweetgum (Liquidambar orientalis), and valerian (Valeriana wallichii) essential oils against pine wood nematode (Bursaphelenchus xylophilus). J Agric Food Chem, 56, 7316-20. https://doi.org/10.1021/jf800780f
  13. Kumar R, Prakash O, Pan AK, et al (2009). Compositional variations and anthelmentic activity of essential oils from rhizomes of different wild populations of Acorus calamus L. and its major component, beta-Asarone. Nat Prod Commun, 4, 275-8.
  14. Kuribayashi K, Mayes PA, El-Deiry, W S (2006). What are caspases 3 and 7 doing upstream of the mitochondria? Cancer Biol Ther, 5, 763-5. https://doi.org/10.4161/cbt.5.7.3228
  15. Lee JY, Yun BS, Hwang BK (2004). Antifungal activity of betaasarone from rhizomes of Acorus gramineus. J Agric Food Chem, 52, 776-80. https://doi.org/10.1021/jf035204o
  16. Lee SH, Kim KY, Ryu SY, et al (2010). Asarone inhibits adipogenesis and stimulates lipolysis in 3T3-L1 adipocytes. Cell Mol Biol (Noisy-le-grand), 56, OL1215-22.
  17. Lee YS, Kim J, Lee SG, et al (2008). Fumigant antifungal activity of essential oil components from Acorus gramineus against three phytopathogenic fungi. Z Naturforsch C, 63, 503-6.
  18. Li WY, Chan SW, Guo DJ, et al (2009). Water extract of Rheum officinale Baill. induces apoptosis in human lung adenocarcinoma A549 and human breast cancer MCF-7 cell lines. J Ethnopharmacol, 124, 251-6. https://doi.org/10.1016/j.jep.2009.04.030
  19. Liao JF, Huang SY, Jan YM, et al (1998). Central inhibitory effects of water extract of Acori graminei rhizoma in mice. J Ethnopharmacol, 61, 185-93. https://doi.org/10.1016/S0378-8741(98)00042-7
  20. Liu J, Li C, Xing G, et al (2010). Beta-asarone attenuates neuronal apoptosis induced by Beta amyloid in rat hippocampus. Yakugaku Zasshi, 130, 737-46. https://doi.org/10.1248/yakushi.130.737
  21. Liu MJ, Wang Z, Li HX, et al (2004). Mitochondrial dysfunction as an early event in the process of apoptosis induced by woodfordin I in human leukemia K562 cells. Toxicol Appl Pharmacol, 194, 141-55. https://doi.org/10.1016/j.taap.2003.08.017
  22. Machover D (1997). A comprehensive review of 5-fluorouracil and leucovorin in patients with metastatic colorectal carcinoma. Cancer, 80, 1179-87. https://doi.org/10.1002/(SICI)1097-0142(19971001)80:7<1179::AID-CNCR1>3.0.CO;2-G
  23. McCullough KD, Martindale JL, Klotz LO, et al (2001). Gadd153 sensitizes cells to endoplasmic reticulum stress by down-regulating Bcl2 and perturbing the cellular redox state. Mol Cell Biol, 21, 1249-59. https://doi.org/10.1128/MCB.21.4.1249-1259.2001
  24. Meiyanto E, Hermawan A and Anindyajati (2012). Natural products for cancer-targeted therapy: citrus flavonoids as potent chemopreventive agents. Asian Pac J Cancer Prev, 13, 427-36. https://doi.org/10.7314/APJCP.2012.13.2.427
  25. Parkin DM, Bray F, Ferlay J, et al (2005). Global cancer statistics, 2002. CA Cancer J Clin, 55, 74-108. https://doi.org/10.3322/canjclin.55.2.74
  26. Qiu D, Hou L, Chen Y, et al (2011). Beta-asarone inhibits synaptic inputs to airway preganglionic parasympathetic motoneurons. Respir Physiol Neurobiol, 177, 313-9. https://doi.org/10.1016/j.resp.2011.05.010
  27. Reed JC (1998). Bcl-2 family proteins. Oncogene, 17, 3225-36.
  28. Ron D, Walter P (2007). Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol, 8, 519-29. https://doi.org/10.1038/nrm2199
  29. Schroder M, Kaufman RJ (2005). ER stress and the unfolded protein response. Mutat Res, 569, 29-63. https://doi.org/10.1016/j.mrfmmm.2004.06.056
  30. Sugimoto N, Goto Y, Akao N, et al (1995). Mobility inhibition and nematocidal activity of asarone and related phenylpropanoids on second-stage larvae of Toxocara canis. Biol Pharm Bull, 18, 605-9. https://doi.org/10.1248/bpb.18.605
  31. Suzuki Y, Imai Y, Nakayama H, et al (2001). A serine protease, HtrA2, is released from the mitochondria and interacts with XIAP, inducing cell death. Mol Cell, 8, 613-21. https://doi.org/10.1016/S1097-2765(01)00341-0
  32. Thornberry NA, Lazebnik Y (1998). Caspases: enemies within. Science, 281, 1312-6. https://doi.org/10.1126/science.281.5381.1312
  33. Wong RS (2011). Apoptosis in cancer: from pathogenesis to treatment. J Exp Clin Cancer Res, 30, 87. https://doi.org/10.1186/1756-9966-30-87
  34. Wu HB, Fang YQ (2004). Pharmacokinetics of beta-asarone in rats. Yao Xue Xue Bao, 39, 836-8.
  35. Zou DJ, Wang G, Liu JC, et al (2011). Beta-asarone attenuates beta-amyloid-induced apoptosis through the inhibition of the activation of apoptosis signal-regulating kinase 1 in SH-SY5Y cells. Pharmazie, 66, 44-51.

Cited by

  1. Inactivated Sendai Virus Strain Tianjin Induces Apoptosis in Human Breast Cancer MDA-MB-231 Cells vol.15, pp.12, 2014, https://doi.org/10.7314/APJCP.2014.15.12.5023
  2. Luteolin, a Bioflavonoid Inhibits Colorectal Cancer through Modulation of Multiple Signaling Pathways: A Review vol.15, pp.14, 2014, https://doi.org/10.7314/APJCP.2014.15.14.5501
  3. Distinct Pro-Apoptotic Properties of Zhejiang Saffron against Human Lung Cancer Via a Caspase-8-9-3 Cascade vol.15, pp.15, 2014, https://doi.org/10.7314/APJCP.2014.15.15.6075
  4. Mitochondria-mediated Apoptosis in Human Lung Cancer A549 Cells by 4-Methylsulfinyl-3-butenyl Isothiocyanate from Radish Seeds vol.15, pp.5, 2014, https://doi.org/10.7314/APJCP.2014.15.5.2133
  5. -Asarone and Their Anticancer Activity vol.2014, pp.2090-2077, 2014, https://doi.org/10.1155/2014/835485
  6. A herbal medicine for Alzheimer's disease and its active constituents promote neural progenitor proliferation vol.14, pp.5, 2015, https://doi.org/10.1111/acel.12356
  7. An in silico Appraisal to Identify High Affinity Anti-Apoptotic Synthetic Tetrapeptide Inhibitors Targeting the Mammalian Caspase 3 Enzyme vol.15, pp.23, 2015, https://doi.org/10.7314/APJCP.2014.15.23.10137
  8. Bufalin Induces Mitochondrial Pathway-Mediated Apoptosis in Lung Adenocarcinoma Cells vol.15, pp.23, 2015, https://doi.org/10.7314/APJCP.2014.15.23.10495
  9. Inhibitory Effects of Phenolic Alkaloids of Menispermum Dauricum on Gastric Cancer in Vivo vol.15, pp.24, 2015, https://doi.org/10.7314/APJCP.2014.15.24.10825
  10. β-asarone inhibits gastric cancer cell proliferation vol.34, pp.6, 2015, https://doi.org/10.3892/or.2015.4316
  11. Hepatotoxic potential of asarones: in vitro evaluation of hepatotoxicity and quantitative determination in herbal products vol.6, pp.1663-9812, 2015, https://doi.org/10.3389/fphar.2015.00025
  12. β-Asarone Induces Apoptosis and Cell Cycle Arrest of Human Glioma U251 Cells via Suppression of HnRNP A2/B1-Mediated Pathway In Vitro and In Vivo vol.23, pp.5, 2018, https://doi.org/10.3390/molecules23051072