Cytotoxicities of Hydrolyzed Crude Laminaran from Eisenia bicyclis on the SNU-1, HeLa and SW Cells

대황으로부터 추출한 crude laminaran 가수분해물의 암 세포독성

  • Published : 2006.12.31

Abstract

We investigated the effects on the cytotoxicity against several cancer cells of the hydrolysis and molecular weight fractionation of crude laminaran from E. bicyclis, a brown seaweed collected from Uleung island in Korea, was extracted with boiling water and then crude laminaran was prepared by ethanol precipitation of extract obtained after elimination of calcium alginate by calcium chloride. Crude laminaran was hydrolyzed by enzyme (Econase CE), acid (0.1 N HCl) and autoclaving ($121^{\circ}C$, 180 min), and the molecular weight fractions by ultrafiltration to generate molecular weight fractions. Total sugar and sulfate contents of hydrolyzed laminaran were 72.3 and 3.5% (enzyme hydrolysate), 68.5 and 3.0% (acid hydrolysate), 70.2 and 3.2% (autoclaved), and monosaccharides of which consisted of glucose (74.7-78.5%), mannose (9.9-11.5%), galactose (8.5-9.6%) and fucose (3.1-4.5%), respectively. When the cytotoxicity of hydrolyzed laminaran on SNU-1, HeLa and SW cells was evaluated by MTT assay, growth-inhibitory activity of the enzyme hydrolysate against cancer cells was higher than that of acid hydrolysate or autoclaved laminaran. Furthermore, the fraction at a molecular weight range of 10 to 50 kDa revealed higher anti-proliferative activities. The $IC_{50}$ values of 10-50 kDa fraction at a molecular weight range of 10 to 50 kDa revealed higher anti-proliferative activities. The $IC_{50}$ values of 10-50 kDa fractions on SNU-1, HeLa and SW cells were 60.4, 58.6 and 53.9 ${\mu}g/mL$ for enzymatic hydrolysate, 75.6, 73.5 and 77.4 ${\mu}g/mL$ for acid hydrolysate, and 61.7, 68.2 and 60.8 ${\mu}g/mL$ for autoclaved, respectively.

References

  1. Jung SE, Bae JH. The effect of Prunus mume extracts on the growth of Hepg 2 and HeLa cell lines. Korean J. Food Nutr. 35: 439-445 (2002)
  2. Kim SS. A study on the related factors of Korean cancer-out-breaks. J. Korean Soc. Health Statistics. 23: 1-16 (1998)
  3. Doll R, Peto R. The causes of cancer. J. Natl. Cancer Inst. 66: 1191-1308 (1981)
  4. Hebert JT, Barone J, Reddy MM, Backllund JC. Natural killer cell activity in dietary fat intervention trial. Clin. Immunol. Immunopathol. 54: 103-106 (1990) https://doi.org/10.1016/0090-1229(90)90010-N
  5. Goldin AI, Kline ZP, Sofina ZP, Syrkin AB. Academic Press, New York. USA. Experimental evaluation of antitumor drugs in the USA and USSR clinical correlation. NIH. pp. 33-44 (1980)
  6. Kamiya H. Endo Y. Uchida A, Raj U. Antitumor activity of the macromolecular fraction from a Fihan tunicate Didemnum carians. Nippon Suisan Goldmishi. 53: 493-504 (1987) https://doi.org/10.2331/suisan.53.493
  7. Sasaki T, Uchida A, Takasuka N, Tachihana Y, Nakamich K, Endo Y, Kamiya H. Antitumor activity and immunomodulatory effect of glycoprotein fraction from scallop, Patinopecten Yessonensis. Nippon Suisan Gokkaishi. 53: 267-273 (1987) https://doi.org/10.2331/suisan.53.267
  8. King CY., Shim MJ, Choi EC, Lee YN, Kim BK. Studies on antitumor components of Korean Basidiomycetes. Mycelial culture and antitumor components of Ganoderma lucidum. Korean Biochem. J.14: 100-109 (1981)
  9. Kim BK, Park EK, Shim M.l. Studies on constituents of higher fungi of Korea (XXIV), antineoplastic activities of coriolus versicolor (lex fr) Qule. Pleurotus ostreatus (Fr) kummer and lentinus edodes (Berk) sing. Arch. Pharm. Res. 2: 145-151 (1979) https://doi.org/10.1007/BF02959042
  10. Ha YL, Michael WP. Naturally-occuring novel anticarcinogens: Conjugated dienoic derivatices of linoleic acid (CLA). Korean J. Food Nutr. 20: 401-407 (1991)
  11. Miyzzaki T. Nishijima M. Structural examination of a water soluble antitumor polysaccharide of Ganoderma lucidum. Chem. Pharm. Bull. 29: 3611-3616 (1981) https://doi.org/10.1248/cpb.29.3611
  12. Franz G. Polysaccharides in phamacy: current applications and future concepts. Planta. Med. 55: 493-497 (1989) https://doi.org/10.1055/s-2006-962078
  13. Purnell D, Bartlet G, Kreidet J, Biro T. Corynebacterium parvum and cyclophosphamides as combination treatment for a mutine mammary adenocar cinoma. Cancer Res. 37: 1137-1140 (1977)
  14. Bjorsson S, Takida H, Combination chemotheraphy plus methanol extracted residue of Bacillus Calmette-Guerin or Corynebacterium parum in stage III lung cancer. Cancer Treat. Rep. 62: 505-510 (1978)
  15. Lee KH, Lee JW, Han MD, Jeong H, Kim YI, Oh DW. Correlation between anticomplementary and antitumor activity of the crude polysaccaride from Ganoderma lucidum IY009. Korean J. Appl. Mirobiol. Biotechnol. 22: 45-51 (1994)
  16. Sugiura M, Ito H. Toxicological studies of Ganoderma lucidum Karst. Tokyo Yakka Daigaku Kenkyu Nempo. 27: 722-733 (1977)
  17. Suzuki I, Hashimoto K, Oikawa S, Sato K, Osawa M, Yadomae T. Antitumor and immunomodulaton activities of a ${\beta}$-glucan obtain from liquid-cultured Grifola flondosa. Chem. Pharm. Bull. 37: 410-413 (1980)
  18. Choi DM, Kim DS, Lee DS, Kim HR, Pyeun JH. Trace components and functional saccharides in seaweed-1. J. Korean Fish Soc. 28: 49-59 (1995)
  19. Cho DM, Kim DS, Lee DS, Kim HR, Pyeun JH. Trace components and functional saccharides in seaweed-2. J. Korean Fish Soc. 28: 270-278 (1995)
  20. Kim DS, Park YH. Uronic acid composition block structure and some related properies of alginic acid. J. Korean Fish Soc. 18: 29-36 (1985)
  21. Tatiana NZ, Nataliya MS, Irina SP, Vladimir VI, Andrey SS, Elena VS, Lyudmila AE. A new procedure for the separation of water-soluble polysaccharides from brown seaweeds. Carbohydrate Res. 322: 32-39 (1999) https://doi.org/10.1016/S0008-6215(99)00206-2
  22. Kim YM, Choi YS, Park JH. Purification and chemical characterisation of laminaran from Eisenia bicyclis in Korea. J. Korean Soc. Food Sci. Nutr. 35: 78-86 (2006) https://doi.org/10.3746/jkfn.2006.35.1.078
  23. Somogyi JN. Notes on sugar detrmination. J. Biochem. 195: 19-23 (1962)
  24. Dubois M. Gills KA, Hamilton JK, Rebers PA, Smith F. Colorimetric method for the determination of sugars and related substances. Anal. Chem. 28: 350-352 (1956) https://doi.org/10.1021/ac60111a017
  25. Lowry OH, Rosebrough NJ, Farr L, Rindall RJ. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193: 256-275 (1951)
  26. Dodgson KS, Price RG. A note on the determination of the ester sulphate content of sulphated polysaccharides. J. Biochem. 84: 106-107 (1962) https://doi.org/10.1042/bj0840106
  27. Furneaux RH, Miller IJ, Stevenson TT. Agaroids from New Zealand members of the Gracilariaceae (Gracilariales. Rhodophyta)- a novel dimethylated agar. Hydrobiologia 645: 204-205 (1990)
  28. Charmichael J, Degraff EG, Gazdaar AF, minna JD, Michell JB. Evaluation of a tetrazolium based semiautomated colorimetric assay, assessment of chemosensitivy testing. Cancer Res. 47: 936-943 (1987)
  29. Tatiana NZ, Natalia MS, Irina VN, Andrey SS, Pavel AL, Ludmila AE. Inhibition of complement activation by water-soluble polysaccharides of some far-eastern brown seaweeds. Comparative Biochemistry and Physiology Part C 126: 209-215 (2000)
  30. Fischer SM, Leyton LJ, Lee mL, Locnislcar M, Belury MA, Maldve RE. Differential effects of dietary linoleic acid on mouse skin-tumor promotion and mammary carcinogenesis. Cancer Res. 52: 2049-2056 (1992)
  31. Yang KH, Yang JH, Ryu BH. Antitumor effects of extracts obtained from Dacdalea dickinsii. Korean J. Appl. Microbiol. Biotechnol. 25: 178-182 (1997)
  32. Song JH, Lee HS, Hwang JK, Han JW, Ro JG, Keum DH, Park KM. Physiological activity of Sarcodon aspratus extracts. Korean J. Food Sci. Ani. Res. 23: 172-179 (2003)
  33. Hwang WI, Sohn JW. Comparative study on the cytotoxic activities of red ginseng of Korea and China. Korean J. Ginseng Sci. 17: 196-200 (1993)
  34. LaVecchia C, Negri E, Decarli A, D'Avanzo B, Franceschi S. Risk factors for hepatocellular carcinoma in northern Italy. Int. J. Cancer 42: 872-876 (1995)
  35. Ziegler RG, Jones CJ, Brinton LA. Diet and risk of in situcervical cancer among white women in the United States. Cancer Causes Control 2: 17-29 (1991) https://doi.org/10.1007/BF00052357