차가버섯과 어성초 함유 발효 조성물이 인체 위암 AGS 및 대장암 HCT-15 세포 생육에 미치는 영향

Effect of Fermented Compositions Containing Inonotus obliquus with Houttuynia cordata on Growth of Human AGS Gastric and HCT-15 Colon Cancer Cells

  • 차재영 (동아대학교 응용생명공학부) ;
  • 전병삼 (경상대학교 의과대학 미생물학교실) ;
  • 박정원 (경상대학교 의과대학 미생물학교실) ;
  • 문재철 ((주)홍재그린) ;
  • 조영수 (동아대학교 응용생명공학부)
  • Cha, Jae-Young (Dept. of Biotechnolgy, College of Natural Resources and Life Science, Dong-A University) ;
  • Jeon, Beong-Sam (Dept. of Microbiology, College of Medicine, Gyeongsang National University) ;
  • Park, Jeong-Won (Dept. of Microbiology, College of Medicine, Gyeongsang National University) ;
  • Moon, Jae-Chul (Hong Jae Green Co., Ltd.) ;
  • Cho, Young-Su (Dept. of Biotechnolgy, College of Natural Resources and Life Science, Dong-A University)
  • 발행 : 2004.06.30

초록

어성초 분말을 포함하는 차가버섯 발효 조성물의 물 추출물이 인체 위암 세포 AGS 및 대장암 세포 HCT-15, 그리고 정상세포 NIH3T3 fibroblast의 세포 생육에 미치는 영향을 검토하였다. 세포독성 실험은 세포수 count와 MTT 방법으로 측정하였다. 어성초 분말을 포함하는 차가버섯 발효 조성물의 제조는 차가버섯과 어성초 분말을 혼합하고, 여기에 대두 발효 미생물원을 다시 혼합시켜 습도 $50{\sim}60%$,온도 $30{\sim}37^{\circ}C$에서 30일 정도 발효시킨 후 건조 시킨 분말에서 5% 물 추출물을 얻어 동결건조 시켜 실험에 제공하였다. MTT assay 방법에서 차가버섯 발효 조성물의 수용성 추출물 0.16, 0.4, 0.8, 1.6 및 4.0 mg/ml 첨가 농도에서 AGS 세포 생육은 13, 25, 40, 67 및78% 억제 되었으며, HCT-15세포 생육은 22, 40, 50, 69및 76%씩 각각 억제되었다. 그러나 동일한 실험조건에서 정상 세포수 NIH3T3은 86% 이상의 생존율을 나타내었다. 차가버섯 발효 조성물의 수용성 추출물은 인체 대장암 세포주 HCT-15와 위암 세포수 AGS에 대해 생육억제 작용이 강한 반면, 정상세포 NIH3T3에 대해서는 세포 독성을 거의 나타내지 않아 가장 바람직한 암예방 또는 항암식품 개발가능성을 제시하였다.

This study was performed to investigate the inhibitory effect of the water-extract from fermented compositions containing Inonotus obliquus with added Houttuynia cordata on the growth of either human AGS gastric and HCT-15 colon cancer cells or NIH3T3 normal mouse fibroblast cells. Cytotoxic activity on cancer cells was investigated by viable cell count, MTT assay and morphological observation. Mixtures of Inonotus obliquus with added Houttuynia cordata were fermented at $30{\sim}37^{\circ}C$, $50{\sim}60%$ humidity for 30 days, extracted with water, freeze dried, powered, and then dissolved in water for the experiment. In MTT assay, the fermented compositions exhibited inhibitory effects of 13, 25, 40, 67 and 78% for AGS and 22, 40, 50, 69 and 76% for HCT-15 at 0.16, 0.4, 0.8, 1.6 and 4.0 mg/ml, respectively. However, normal NIH3T3 cells were exhibited 86% survival under the same experimental condition. Fermented compositions showed highly inhibitory effect against human cancer cell line HCT-15 and AGS, but not on normal cell line NIH3T3.

키워드

참고문헌

  1. Kier, L. (1961) Triterpenes of Poria obliqua. J. Pharm. Sci. 50 471-474 https://doi.org/10.1002/jps.2600500605
  2. Shivrina, A. N. (1967) Chemical characteristics of compounds extracted from Irwnotus obliquus. Chem. Abstr. 66, 17271-17279
  3. Kahlos, K., Kangas, L. and Hitunen, R., (1987) Antitumor activity of some compounds and fractions from an n-hexane extract of Inonotus obliquus. Acta Pharm. Fenn 96, 33-40
  4. Mizuno, T., Zhuang, C., Abe, K., Okamoto, H., Kiho, T., Ukai, S., Leclerc, S. and Meijer, L. (1999) Antitumor and hypohlycemic activities of polysaccharides from the Sclerotia and Mycelia of Inonotus obliquus (Pers.: Fe.) Pll. (Aphyllophoromycetideae). Int. J. Med Mushrooms 1, 301-316 https://doi.org/10.1615/IntJMedMushr.v1.i4.20
  5. Wasser, S. P. (2002) Medicinal mushrooms as a source of antitumor and imrnunomodulating polysaccharides. Appl. Microbiol. Biotechnol. 60, 258-274
  6. Mizuno, T., Minato, K., Ito, H., Kawade, M., Terai, H. and Tsuchida, H. (1999) Antitumor polysaccharide from the mycelium of liquid-cultured Agricus blazei Murrill. Biochem. Mol. BioI. Int. 47, 707-714
  7. Ham, S. S., Oh, S. W, Kim, Y. K., Shin, K. S., Chang, K. Y. and Chung, G. H. (2003) Antimutagenic and cytotoxic effects of ethanol extract from the Inonotus obliquus. J Korean Soc. Food Sci. Nutr. 32, 1088-1094 https://doi.org/10.3746/jkfn.2003.32.7.1088
  8. Murakami, C., Hirakawa, Y., lnui, H. and Nakano, Y. (2002) Effect of tea catechins on cellular lipid peroxidation and cytotoxicity in HepG2 cells. Biosci. Biotechnol .Biochem. 66, 1559-1562
  9. Vemon, E. S. and Gary, J. (2000) Comparative chemopreventive mechanism of green tea, black tea and selected polyphenol extracts measured by in vitro bioassay. Carcinogenesis 21, 63-67 https://doi.org/10.1093/carcin/21.1.63
  10. Song, J. H., Kim, M. J., Kwon, H. D. and Park, I. H. (2003) Antimicrobial activity of fractional extracts from Houttuynia cordata root. J Korean Soc. Food Sci. Nutr.32, 1055-1058
  11. Sung, N. J., Lee, S. J., Shin, J. H. and Kim, H. S. (1998) The effect of feeding juice and powder of Houttuynia cordata Thunb on serum lipids in rats. J. Korean Soc. Food Sci. Nutr. 27, 1223-1229
  12. Hayashi, K., Kamiya, M. and Hayashi, T. (1995) VIrucidal effects of the stearm distillate from Houttuynia cordata Thunb and its components on HSV-1, influenza virus and HIV. Planta Med. 61, 237-240 https://doi.org/10.1055/s-2006-958063
  13. Kim, S. K., Ryu, S. Y., No, J., Choi, S. U. and Kim, Y. S. (2001) Cytotoxic alkaloids from Houttuynia cordata. Arch. Pharm. Res. 24, 518-521
  14. Lee, S. T., Lee, Y. H., Choi, Y. J., Shon, G. M., Lee, H. J. and Heo, J. S. (2002) Composition of quercetin and soluble tannin in Houttuynia cordata Thunb according to growth stages and plant parts. Korean J. Medicinal Corp Sci. 10, 12-16
  15. Ok, M., Kim, M. S., Seo, W. S. and Cha, J. Y., Cho, Y. S. (2000) Characteization of extracellular protease of Bacillus sp. WRD-l isolated from soil. Kor. J. Appl. Microbiol. Biotechnol. 28, 329-333
  16. Yotsumoto, H., Yanagita, T., Yamamoto, K., Ogawa, Y., Cha, J. Y. and Mori, Y. (1997) Inhibitory effects of Oren-Gedoku-to and its components on cholesteryl ester synthesis in cultured human hepatocyte HepG2 cells: Evidence from the cultured HepG2 cells and in vitro assay ACAT. Planta Med. 63, 141-145 https://doi.org/10.1055/s-2006-957631
  17. Yanagita, T., Hara, E., Yotsumoto, H., Rahman, S. M., Han, S. Y., Cha, J. Y. and Yamamoto, K .(1999) NK-104, a potent new 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, enhances posttranslational catabolism of apolipoprotein B-lOO and inhibits secretion of apolipoprotein B-100 and triacylglycerols from HepG2 cells. Curr. Ther. Res. 60, 423-434 https://doi.org/10.1016/S0011-393X(99)80021-6
  18. Kim, S. W. (1998) Studies on anti-microbial and anti-cancer function of polysaccharide extracted Ganoderma lucidum. J. Korean Soc. Food Sci. Nutr. 27, 1183-1188
  19. Li, X., Rong, J., Wu, M. and Zeng, X. (2003) Anti-tumor effect of polysaccharide from Grifola frondosa and its int1uence on immunological function. Zhong Yao Cai. 26, 31-32
  20. Goro, C. Jnuli, H., Yukiko, Y., Yoshiko, A. and Fumoko, F. (1970) Fractionation and purification of the polysaccharides with masked antitumor activity, especially lentinan from Lentinus edodes (Berk.) Sing (an edible mushroom). Cancer Res. 30, 2776-2781
  21. Kawagishi, H., Inagaki, R., Kanao, T., Mizuno, T., Shimura, K., Ito, H., Hagiwara, T. and Hakamura, T. (1989) Fractionation and antitumor activity of the water-insoluble residue of Agricus blazei fruiting bodies. Carbohydr. Res. 186, 267-274 https://doi.org/10.1016/0008-6215(89)84040-6
  22. Song, H. N. and Oh, S. W. (2002) Optimization of extraction and clarification condition for preparation of liquid extract tea from artificially cultivated Phellinus linteus. J. Korean Soc. Food Sci. Nutr. 31, 636-641 https://doi.org/10.3746/jkfn.2002.31.4.636
  23. Ohno, N., Miura, N. N., Nakajima, M. and Yadomae, T. (2000) Antitumor 1,3-beta-glucan from cultured fruit body of Sparassis crispa. Biol. Pharm. Bull. 23, 866-872 https://doi.org/10.1248/bpb.23.866
  24. Suzuki, K, Mikami, T., Okawa, Y., Tokoro, A., Suzuki, S. and Suzuki, M. (1986) Antitumor effect of hexa-N-acetylchitohexaose and chitohexaose. Carbohydr. Res. 151, 403-408 https://doi.org/10.1016/S0008-6215(00)90359-8
  25. Korean National Cancer Center (2002) Annual Report on the Cause of Cancer Statistics
  26. Dnizot, F. D. and Rita, L. (1986) Rapid colorimetric assay for cell growth and survival: Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J. Immunoi. Methods 22, 271-277