Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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In vitro and in vivo Effects of Extracts of Lentinus edodes on Tumor Growth in a Human Papillomavirus 16 Oncogenes-transformed Animal Tumor Model -Apoptosis-mediated Tumor Cell Growth Inhibition-

자궁경부암동물세포에서 표고버섯의 in vitro 및 in vivo 항암효과 -Apoptosis에 의한 종양세포주의 성장억제-

  • Park, Jeong-Min (Department of Medicine, College of Medicine, Catholic University of Daegu) ;
  • Lee, Sung-Hyun (Agriproduct Science Division, National Rural Living Science Institute) ;
  • Kim, Jung-Ok (Department of Medicine, College of Medicine, Catholic University of Daegu) ;
  • Park, Hong-Ju (Agriproduct Science Division, National Rural Living Science Institute) ;
  • Park, Jae-Bok (Department of Medicine, College of Medicine, Catholic University of Daegu) ;
  • Sin, Jeong-Im (Department of Medicine, College of Medicine, Catholic University of Daegu)
  • 박정민 (대구가톨릭대학교 의과대학 의학과) ;
  • 이성현 (농촌생활연구소 농산물가공이용과) ;
  • 김정옥 (대구가톨릭대학교 의과대학 의학과) ;
  • 박홍주 (농촌생활연구소 농산물가공이용과) ;
  • 박재복 (대구가톨릭대학교 의과대학 의학과) ;
  • 신정임 (대구가톨릭대학교 의과대학 의학과)
  • Published : 2004.02.28
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Abstract

Fungal products indirectly mediate anti-tumor effects in vitro and in vivo. To investigate whether Lentinus edodes might possess direct anti-tumor substance, L. edodes was extracted and tested on human papillomavirus (HPV) 16 oncogenes-associated animal tumor cells (TC-1) and in an animal tumor model. Only water extract displayed direct anti-proliferative effects in TC-1 tumor cells in vitro. This inhibition was dose-dependent, and inhibitory concentration ($IC_{50}$) was $800\;{\mu}g/mL$. Fungal extracts also showed growth inhibition to human cervical cancer cells (CaSki and HeLa) similarly to TC-1 tumor cells. When fungal extracts were added at a high dose (1.5 mg/mL), cell growth was inhibited within 6 hr following extract treatment. Cell growth inhibition was blocked by heat treatment, but not by low pH, which is indicative of heat sensitivity of this anti-proliferative substance. Cell growth suppression was mediated by apoptosis, as determined by Annexin V and propidium iodide staining. When challenged with TC-1 cells, direct intratumoral injection of fungal extracts resulted in some positive effect on tumor growth inhibition, as compared to oral delivery. Results suggest that heat labile substance of L. edodes suppresses growth of HPV oncogenes-associated tumor cells through apoptosis.

여러 발표에서 버섯추출물의 간접적 항종양효과에 대해 보고 되고 있다. 본 연구는 표고버섯의 직접적인 종양세포주 성장억제효과를 탐지하기 위하여 휴먼자궁경부암바이러스 타입 16의 종양유전자가 발현된 동물종양세포주(TC-1)를 이용하였다. 여러 추출용매 중 물을 이용한 추출액에서 TC-1 세포주의 성장억제효과를 탐지할 수 있었다. 이들 세포성장저해는 추출물첨가농도에 상응하며 $IC_{50}$는 $800\;{\mu}g/mL$로 계산되었다. 또한, 표고버섯 물 추출물은 휴먼자궁경부암세포주(CaSki and HeLa)에서도 유사한 세포성장억제효과를 보여 주었다. 첨가농도가 1.5 mg/mL에서는 첨가후 6시간째에 벌써 완전한 종양세포주(TC-1)의 성장억제가 탐지되었다. 이들 추출물을 열처리시 종양세포주성장억제가 소실되었으나 pH의 변화에서는 그대로 종양세포주성장억제효과가 탐지되었다. 이는 표고버섯 물 추출물 내에 있는 항종양물질이 열에 민감함을 의미한다. Annexin V 및 propidium iodide 염색에 의해 이들 TC-1종양세포주의 성장 억제효과는 apoptosis에 의해 매개됨을 보여 주었다 동물실험에서 표고버섯 물 추출물의 구강투여시 TC-1유래 종양의 성장 및 생성에 아무 효과를 주지 못하였다. 반면, 형성된 종양에 직접 주사시 종양크기의 감소가 탐지되었다. 요약하면 이들 연구에서 표고버섯 물 추출물에 존재하는 항암물질은 열에 민감하며 apoptosis에 의해 휴먼자궁경부암종양유전자에 의해 종양화된 세포주의 세포성장을 직접 억제하는 기능이 있음을 증명한다.

Keywords

References

  1. Lee BW, Park KM. Anti-tumor activity of protein-bound polysaccharides extracted from mycelia of Lentinus edodes. Korean J. Food Sci. Technol. 30: 665-671 (1998)
  2. Ikekawa T, Uhehara N, Maeda Y, Nakanishi M, Fukuoka F. Antitumor activities of aquous extracts of some edible mushrooms. Cancer Res. 29: 734-735 (1969)
  3. Takuma S, Nobuo T. Further study of the structure of lentinan, antitumor polysaccharides from Lentinus edodes. Carbohydr. Res. 47: 99-104 (1976) https://doi.org/10.1016/S0008-6215(00)83552-1
  4. Park MH, Oh KY, Lee BW. Anti-cancer activity of Lentinus edodes and Pleurotus astreatus. Korean J. Food Sci. Technol. 30: 702-708 (1998)
  5. Dennert G, Tucker D. Antitumor polysaccharide lentinan a T cell adjuvant. J. Natl. Cancer. Inst. 51: 1727-1735 (1973) https://doi.org/10.1093/jnci/51.5.1727
  6. Hamuro G, Wagner H. $\beta$-1,3-glucan-mediated augmentation of alloreactive murine cytotoxic T lymphocytes in vivo. Cancer Res. 38: 3080-3088 (1978)
  7. Chihara G. Immune modulation agents and their mechanisms (lentinan, a T cell oriented immunopotentiator). Basel 19: 409-436 (1985)
  8. Nakano H, Namatame K, Nemoto H, Motohashi H, Nishiyama K, Kumada K. A multi-institutional prospective study of lentinan in advanced gastric cancer patients with unresectable and recurrent diseases: effect on prolongation of survival and improvement of quality of life. Hepatogastroenterology 46: 2662-2668 (1999)
  9. Kimura Y, Iijima S, Kato T, Tsujie M, Naoi Y, Hayashi T, Tanigawa T, Yamamoto H, Kurokawa E, Kikkawa N, Matsuura N. Usefulness of TS-1 and lentinan combination immunochemotherapy in advanced or recurrent gastric cancer-pilot study aiming at a randomized trial. Gan To Kagaku Ryoho 30: 1125-1130 (2003)
  10. Ahn WS, Kim DJ, Chae GT, Lee JM, Bae SM, Sin JI, Kim YW, Namkoong SE, Lee IP. Natural killer cell activity and quality of Life were improved by consumption of a mushroom extract, Agaricus Blazei Murill Kyowa in gynecological cancer patients undergoing chemotherapy. Int. J. Gynecol. Cancer. 61: in press (2004)
  11. Kim TY, Myoung HJ, Kim JH, Moon IS, Kim TG, Ahn WS, Sin JI. Both E7 and CpG-ODN are required for protective immunity against challenge with human papillomavirus 16 (E6/E7)-immortalized tumor cells: Involvement of CD4+ and CD8+ T cells in protection. Cancer Res. 62: 7234-7240 (2002)
  12. Ahn WS, Bae SM, Kim TY, Kim TG, Lee JM, Namkoong SE, Kim CK, Sin JI. A therapy modality using recombinant IL-12 adenovirus plus E7 protein in a human papillomavirus 16 E6/E7- associated cervical cancer animal model. Hum. Gene Ther. 14: 1389-1399 (2003) https://doi.org/10.1089/104303403769211619
  13. Kim TW, Hung CF, Ling M, Juang J, He L, Hardwick JM, Kumar S, Wu TC. Enhancing DNA vaccine potency by coadministration of DNA encoding antiapoptotic proteins. J. Clin. Invest. 112: 109-117 (2003) https://doi.org/10.1172/JCI200317293
  14. Ahn WS, Han YJ, Bae SM, Kim TH, Rho MS, Lee JM, Namkoong SE, Park YS, Kim CK, Sin JI. Differential suppression of human cervical cancer cell growth by adenovirus delivery of p53 in vitro: arrest phase of cell cycle is dependent on cell line. Jpn. J. Cancer Res. 93: 1012-1019 (2002) https://doi.org/10.1111/j.1349-7006.2002.tb02478.x
  15. Romijn JC, Verkoelen CF, and Schroeder FH. Application of the MTT assay to human prostate cancer cell lines in vitro: establishment of test conditions and assessment of hormone-stimulated growth and drug-induced cytostatic and cytotoxic effects. Prostate 12: 99-110 (1988) https://doi.org/10.1002/pros.2990120112
  16. Li X, Ding X, Adrian TE. Arsenic trioxide induces apoptosis in pancreatic cancer cells via changes in cell cycle, caspase activation, and GADD expression. Pancreas 27: 174-179 (2003) https://doi.org/10.1097/00006676-200308000-00011
  17. Ahn WS, Huh SW, Bae SM, Namkoong SE, Park YS, Kim CK, Sin JI. A major constituent of tea, (-)-epigallocatechin-3-gallate inhibits cervical cancer cell growth through apoptosis, cell cycle arrest and regulation of gene expression. DNA Cell Biol. 22: 217-224 (2003) https://doi.org/10.1089/104454903321655846
  18. Park KM, Lee BW. Extraction and purification of antitumor protein-bound polysaccharides from mycelia of Lentinus edodes. Korean J. Food Sci. Technol. 30: 1236-1242 (1998)
  19. Rhee YK, Han MJ, Park SY, Kim DH. In vitro and in vivo antitumoractivity of the fruit body of Phellinus linteus. Korean J. Food Sci. Technol. 32: 477-480 (2000)