Browse > Article

Apoptosis of Human Hepatocarcinoma (HepG2) and Neuroblastoma (SK-N-SH) Cells Induced by Polysaccharides-Peptide Complexes Produced by Submerged Mycelial Culture of an Entomopathogenic Fungus Cordyceps sphecocephala  

Oh, Jung-Young (Department of Biotechnology, Daegu University)
Baek, Yu-Mi (Department of Biotechnology, Daegu University)
Kim, Sang-Woo (Department of Biotechnology, Daegu University)
Hwang, Hye-Jin (Department of Biotechnology, Daegu University)
Hwang, Hee-Sun (Department of Biotechnology, Daegu University)
Lee, Sung-Hak (Department of Biotechnology, Daegu University)
Yun, Jong-Won (Department of Biotechnology, Daegu University)
Publication Information
Journal of Microbiology and Biotechnology / v.18, no.3, 2008 , pp. 512-519 More about this Journal
Abstract
Three different polysaccharide-peptide complexes (PPC, named as Fr-I, Fr-II, and Fr-III) were produced by submerged mycelial culture of an entomopathogenic fungus Cordyceps sphecocephala, and their anticancer activities were investigated in human hepatocarcinoma (HepG2) and neuroblastoma (SK-N-SH) cells. The highest inhibitory effects of PPC on both HepG2 and SK-N-SH cells were achieved with Fr-I, whereas Fr-III with low molecular mass showed lower inhibition effects. Interestingly, the inhibitory effects of the three fractions were increased after protease digestion, suggesting that the inhibitory effects resulted mainly from the carbohydrate moiety, at least in the case of Fr-II and Fr-III, of PPC. The results of DNA fragmentation in PPC-induced apoptotic cells were confirmed by both DNA ladder assay and comet assay. Our investigation also showed that PPC-induced apoptosis of both cancer cells was associated with intracellular events including DNA fragmentation, activation of caspase-3, and modulation of Bcl-2 and Bax. We conclude that PPC has potential as a novel therapeutic agent for the treatment of both HepG2 and SK-N-SH cancer cells without any cytotoxicity against normal cells.
Keywords
Anticancer activity; apoptosis; comet assay; Cordyceps sphecocephala; HepG2; SK-N-SH;
Citations & Related Records
Times Cited By KSCI : 4  (Citation Analysis)
Times Cited By Web Of Science : 0  (Related Records In Web of Science)
연도 인용수 순위
1 Cooper, J. D., T. M. Michaelidis, G. Z. Tzimagiorgis, M. Sendtner, M. Meyer, and H. Thoenen. 1996. Cholinergic neuron-specific overexpression of Bax in ChAT Bax transgenic mice. Soc. Neurosci. 22: 570
2 Gorczyca, W., K. Bigman, A. Mittelman, T. Ahmed, J. Gong, M. R. Melamed, and Z. Darzynkiewicz. 1993. Induction of DNA strand breaks associated with apoptosis during treatment of leukemias. Leukemia 7: 659-670
3 Kim, H. M., S. W. Kim, H. J. Hwang, M. K. Park, Y. A. G. Mahmoud, J. W. Choi, and J. W. Yun. 2006. Influence of agitation intensity and aeration rate on the production of antioxidative exopolysaccharides from submerged mycelial culture of Ganoderma recinaceum. J. Microbiol. Biotechnol. 16: 1240-1247   과학기술학회마을
4 Kim, S. W., H. J. Hwang, C. P. Xu, J. M. Sung, J. W. Choi, and J. W. Yun. 2003. Optimization of submerged culture process for the production of mycelial biomass and exo-polysaccharides by Cordyceps militaris C738. J. Appl. Microbiol. 94: 120-126
5 Kobayashi, T., S. Ruan, K. Clodi, K. O. Kliche, H. Shiku, M. Andreeff, and W. Zhang. 1998. Overexpression of Bax gene sensitizes K562 erythroleukemia cells to apoptosis induced by selective chemotherapeutic agents. Oncogene 16: 1587-1591   DOI   ScienceOn
6 Martin, S. J. and D. R. Green. 1995. Protease activation during apoptosis: Death by a thousand cuts? Cell 82: 349-352   DOI   ScienceOn
7 Ohmori, T., K. Tamura, K. Fukui, G. Kawanishi, M. Mitsuyama, K. Nomoto, and T. Miyazaki. 1989. Isolation of galactosaminoglycan moiety (CO-N) from protein-bound polysaccharides of Cordyceps ophioglossoides and its effects against murine tumors. Chem. Pharm. Bull 37: 1019-1022   DOI   ScienceOn
8 Surenjav, U., L. Zhang, X. Xu, X. Zhang, and F. Zeng. 2006. Effects of molecular structure on antitumor activities of (13)-$\beta$-D-glucans from different Lentinus edodes. Carbohydr. Polym. 63: 97-104   DOI   ScienceOn
9 Tu, H., S. C. Jacobs, A. Borkowski, and N. Kyprianou. 1996. Incidence of apoptosis and cell proliferation in prostate cancer: Relationship with TGF-b and bcl-2 expression. Int. J. Cancer 69: 357-363   DOI   ScienceOn
10 Wang, H. X., T. B. Ng, V. E. Ooi, W. K. Liu, and S. T. Chang. 1996. A polysaccharide-peptide complex from cultured mycelia of the mushroom Tricholoma mongolicum with immunoenhancing and anti-tumor activities. Biochem. Cell Biol. 71: 95-100
11 Yamada, H., H. Kiyohara, J. C. Cyong, and Y. Otsuka. 1985. Studies on the polysaccharides from Angelica acutiloba. Characterization of an anti-complementary arabinogalactan from the roots of Angelica acutiloba Kitakawa. Mol. Immunol. 22: 295-302   DOI   ScienceOn
12 Lim, H. Y., Y. H. Lim, Y. H. Cho, and C. H. Lee. 2006. Induction of apoptosis in the HepG2 cells by HY53, a novel natural compound isolated from Bauhinia forficata. J. Microbiol. Biotechnol. 16: 1262-1268   과학기술학회마을
13 Budihardjo, I., H. Oliver, M. Lutter, X. Luo, and X. Wang. 1999. Biochemical pathways of caspase activation during apoptosis. Annu. Rev. Cell Dev. Biol. 15: 269-290   DOI   ScienceOn
14 Compton, M. M. 1992. A biochemical hallmark of apoptosis: Internucleosomal degradation of the genome. Cancer Met. Rev. 11: 105-119   DOI   ScienceOn
15 Tice, R. R., E. Agurell, D. Anderson, B. Burlinson, A. Hartmann, H. Kobayashi, Y. Miyamae, E. Rojas, J. C. Ryu, and Y. F. Sasaki. 2000. Single cell gel/comet assay: Guidelines for in vitro and in vivo genetic toxicology testing. Environ. Mol. Mutagen. 35: 206-221   DOI   ScienceOn
16 Chen, Y. J., M. S. Shiao, S. S. Lee, and S. Y. Wang. 1997. Effects of Cordyceps sinensis on the proliferation and differentiation of human leukemic U937 cells. Life Sci. 60: 2349-2359   DOI   ScienceOn
17 Mosmann, T. 1983. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J. Immunol. Methods 65: 55-63   DOI
18 Evan, G. I. and K. H. Vousden. 2001. Proliferation, cell cycle and apoptosis in cancer. Nature 411: 342-348   DOI   ScienceOn
19 Fukamachi, Y., Y. Karasaki, T. Sugiura, H. Itoh, T. Abe, K. Yamamura, and K. Higashi. 1998. Zinc suppresses apoptosis of U937 cells induced by hydrogen peroxide through an increase of the Bcl-2/Bax ratio. Biochem. Biophys. Res. Commun. 246: 364-369   DOI   ScienceOn
20 Kumar, S., P. K. Suresh, M. R. Vijayababu, A. Arunkumar, and J. Arunakaran. 2006. Anticancer effects of ethanolic neem leaf extract on prostate cancer cell line (PC-3). J. Ethnopharmacol. 105: 246-250   DOI
21 Yamada, H., K. Ohtani, H. Kiyohara, J. C. Cyung, Y. Otsuka, Y. Ueno, and S. Omura. 1985. Purification and chemical properties of anti-complementary polysaccharide from the leaves of Artemisia princeps. Planta Medica 51: 121-125   DOI   ScienceOn
22 Zhang, M., S. W. Cui, P. C. K. Cheung, and Q. Wang. 2007. Antitumor polysaccharides from mushrooms: A review on their isolation process, structural characteristics and antitumor activity. Trends Food Sci. Technol. 18: 4-19   DOI   ScienceOn
23 Zhu, T., L. Collins, J. Kelly, Z. J. Xiao, S. H. Kim, and C. Y. Chen. 2007. Phellinus linteus activates different pathways to induce apoptosis in prostate cancer cells. Br. J. Cancer 96: 583-590   DOI   ScienceOn
24 Salvesen, G. S. and V. M. Dixit. 1997. Caspases: Intracellular signaling by proteolysis. Cell 91: 443-446   DOI   ScienceOn
25 Bradford, M. M. 1976. A rapid and sensitive method for the quantization of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254   DOI   ScienceOn
26 Koh, J. H., J. M. Kim, U. J. Chang, and H. J. Suh. 2003. Hypocholesterolemic effect of hot-water extract from mycelia of Cordyceps sinensis. Biol. Pharm. Bull. 26: 84-87   DOI   ScienceOn
27 Sakahira, H., M. Enari, and S. Nagata. 1998. Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature 391: 96-99   DOI   ScienceOn
28 Taylor, P. R., G. D. Brown, D. M. Reid, J. A. Willment, L. M. Pomares, S. Gordon, and S. Y. C. Wong. 2002. The $\beta$-glucan receptor, Dectin-1, is predominantly expressed on the surface of cells of the monocyte/macrophage and neutrophil lineages. J. Immunol. 169: 3876-3882   DOI
29 Lazebnik, Y. A., S. H. Kaufmann, S. Desnoyers, G. G. Poirier, and W. C. Earnshaw. 1994. Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature 371: 346-347   DOI   ScienceOn
30 Yin, X. M., Z. Oltvai, and S. J. Korsmeyer. 1994. BH1 and BH2 domains of bcl-2 are required for inhibition of apoptosis and heterodimerization with bax. Nature 369: 321-323   DOI   ScienceOn
31 Liu, F., V. E. C. Ooi, and S. T. Chang. 1995. Anti-tumour components of the culture filtrates from Tricholoma sp. World J. Microbiol. Biotechnol. 11: 486-490   DOI   ScienceOn
32 Kaufmann, S. H. and W. C. Earnshaw. 2000. Induction of apoptosis by cancer chemotherapy. Exp. Cell Res. 256: 42-49   DOI   ScienceOn
33 Kiho, T., K. Ookubo, S. Usui, S. Ukai, and K. Hirano. 1999. Strutural features and hypoglycemic activity of a polysaccharide (CS-F10) from the cultured mycelium of Cordyceps sinensis. Biol. Pharm. Bull. 22: 966-970   DOI   ScienceOn
34 Nakamura, K., Y. Yamaguchi, S. Kagota, K. Shinozuka, and M. Kunitomo. 1999. Activation of in vivo Kupffer cell function by oral administration of Cordyceps sinensis in rats. Jap. J. Pharmacol. 79: 505-508   DOI
35 Kiho, T., A. Yamane, J. Hui, S. Usui, and S. Ukai. 1996. Polysaccharides in fungi XXXVI: Hypoglycemic activity of a polysaccharide (CS-F30) from the cultural mycelium of Cordyceps sinensis and its effect on glucose metabolism in mouse liver. Biol. Pharm. Bull. 19: 294-296   DOI   ScienceOn
36 Park, J. P., Y. M. Kim, S. W. Kim, H. J. Hwang, Y. J. Cho, Y. S. Lee, and J. W. Yun. 2002. Effect of aeration rate on the mycelial morphology and exo-biopolymer production in Cordyceps militaris. Process Biochem. 37: 1257-1262   DOI
37 Yasuhara, S., Y. Zhu, T. Matsui, N. Tipirneni, Y. Yasuhara, M. Kaneki, A. Rosenzweig, and J. A. Martyn. 2003. Comparison of comet assay, electron microscopy, and flow cytometry for detection of apoptosis. J. Histochem. Cytochem. 51: 873-885   DOI   ScienceOn
38 Thornberry, N. A. 1997. The caspase family of cysteine proteases. Br. Med. Bull. 53: 478-490   DOI   ScienceOn
39 Wasser, S. P. 2002. Medicinal mushrooms as a source of antitumor and immuno-modulating polysaccharides. Appl. Microbiol. Biotechnol. 60: 258-274   DOI   ScienceOn
40 Li, N., X. H. Liu, J. Zhou, Y. X. Li, and M. W. Zhao. 2006. Analysis of influence of environmental conditions on ganoderic acid content in Ganoderma lucidum using orthogonal design. J. Microbiol. Biotechnol. 16: 1940-1946   과학기술학회마을
41 Peng, Y., L. Zhang, F. Zeng, and Y. Xu. 2003. Structure and antitumor activity of extracellular polysaccharides from mycelium. Carbohydr. Polym. 54: 297-303   DOI   ScienceOn
42 Oh, J. Y., E. J. Cho, S. H. Nam, J. W. Choi, and J. W. Yun. 2007. Production of polysaccharide-peptide complexes by submerged mycelial culture of an entomopathogenic fungus Cordyceps sphecocephala. Process Biochem. 42: 3352-3362
43 Mizuno, T., P. Yeohlui, T. Kinoshita, C. Zhung, H. Ito, and Y. Mayuzumi. 1996. Antitumor activity and chemical modification of polysaccharides from Nioshimeiji mushroom, Tricholoma giganteum. Biosci. Biotechnol. Biochem. 60: 30-33   DOI   ScienceOn
44 Moradali, M. F., H. Mostafavi, S. Ghods, and G. A. Hedjaroude. 2007. Immunomodulating and anticancer agents in the realm of macromycetes fungi (macrofungi). Int. Immunopharmacol. 7: 701-724   DOI   ScienceOn
45 Tao, Y., L. Zhang, and P. C. K. Cheung. 2006. Physicochemical properties and antitumor activities of water-soluble native and sulfated hyperbranched mushroom polysaccharides. Carbohydr. Res. 341: 2261-2269   DOI   ScienceOn
46 Choi, D. B., J. M. Maeng, J. L. Ding, and W. S. Cha. 2007. Exopolysaccharide production and mycelial growth in an air-lift bioreactor using Fomitopsis pinicola. J. Microbiol. Biotechnol. 17: 1369-1378   과학기술학회마을
47 Oltvai, Z. N., C. L. Milliman, and S. J. Korsmeyer. 1993. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74: 609-619   DOI   ScienceOn
48 Earnshaw, W. C. 1995. Nuclear changes in apoptosis. Curr. Opin. Cell Biol. 7: 337-343   DOI   ScienceOn
49 Matsunaga, K., A. Hosokawa, M. Oohara, N. Sugita, M. Harada, and K. Nomoto. 1998. Direct action of a proteinbound polysaccharide, PSK, on transforming growth factor-$\beta$. Immunopharmacology 40: 219-230   DOI   ScienceOn
50 Jeong, S. C., B. K. Yang, K. S. Ra, M. A. Wilson, Y. Cho, Y. A. Gu, and C. H. Song. 2004. Characteristics of anticomplementary biopolymer extracted from Coriolus versicolor. Carbohydr. Polym. 55: 255-263   DOI   ScienceOn