Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Induction of Apoptotic Cell Death by Aqueous Extract of Cordyceps militaris Through Activation of Caspase-3 in Human Hepatocarcinoma Hep3B Cells

Hep3B 간암세포에서 Caspase-3 활성화를 통한 동충하초 열수추출물의 Apoptosis 유도에 관한 연구

  • Kim, Kyung-Mi (Dept. of Biology, Pusan National University) ;
  • Park, Cheol (Dept. of Biology, Pusan National University) ;
  • Seo, Sang-Ho (Dept. of Oriental Medicine, Dongeui University College of Oriental Medicine) ;
  • Hong, Sang-Hoon (Dept. of Oriental Medicine, Dongeui University College of Oriental Medicine) ;
  • Lee, Won-Ho (Dept. of Biology, Pusan National University) ;
  • Choi, Yung-Hyun (Dept. of Oriental Medicine, Dongeui University College of Oriental Medicine)
  • 김경미 (부산대학교 자연과학대학 생물학과) ;
  • 박철 (부산대학교 자연과학대학 생물학과) ;
  • 서상호 (동의대학교 한의과대학 한의학과) ;
  • 홍상훈 (동의대학교 한의과대학 한의학과) ;
  • 이원호 (부산대학교 자연과학대학 생물학과) ;
  • 최영현 (동의대학교 한의과대학 한의학과)
  • Published : 2008.06.30
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Abstract

Cordyceps militaris is a medicinal fungus which has been used for patient suffering from cancer in Oriental medicine. It was previously reported that C. militaris extracts are capable of inhibiting tumor growth and inducing apoptosis; however, the anti-poliferative effects of human cancer cells have been poorly understood. In this study, to elucidate the anti-cancer mechanisms of human cancer cells by treatment with aqueous extract of C. militaris (AECM), we investigated the anti-proliferative effects of AECM in human hepatocarcinoma Hep3B cells. AECM treatment inhibited the growth of Hep3B cells and induced the apoptotic cell death in a concentration-dependent manner such as formation of apoptotic bodies and increased populations of apoptotic-sub G1 phase. The induction of apoptosis by AECM was connected with a proteolytic activation of caspase-3 and caspase-8. and concomitant degradation of poly (ADP-ribose) polymerase (PARP) and ${\beta}$-catenin proteins. Furthermore, caspase-3 inhibitor, z-DEVD-fmk, significantly inhibited AECM-induced apoptosis demonstrating the important role of caspase-3 in the bserved cytotoxic effect. Taken together, these findings suggest that AECM-induced inhibition of human hepatocarcinoma cell proliferation is associated with the induction of apoptotic cell death via activation of caspase-3 and C. militaris may have therapeutic potential in human cancer.

본 연구에서는 동충하초(C. militaris)의 항암작용 기전 해석을 위하여 Hep3B 간암세포의 apoptosis 유발에 미치는 동충하초 열수추출물(WECM)의 영향을 조사하였으며, apoptosis 조절에 중요한 몇 가지 유전자들의 발현 및 활성 변화를 조사하였다. AECM 처리에 의한 Hep3B 세포의 증식억제는 형태적 변형을 동반한 apoptosis 유도와 연관성이 있음을 DAPI 염색을 통한 apoptotic body 출현의 증가 및 flow cytometry 분석에 의한 sub-G1 기에 속하는 세포 빈도의 증가로 확인하였다. AECM 처리에 의한 apopotosis 유도에서 Bcl-2 family에 속하는 몇 가지 유전자들의 발현은 큰 변화가 없었으나, caspase-3 및 -8의 활성이 매우 높게 증가 되었으며 이는 PARP 및 ${\beta}$-catenin 단백질의 분해와 연관성이 있었다. 또한 caspase-3 선택적 저해제인 z-DEVD-fmk로 caspase-3의 활성을 인위적으로 차단시켰을 경우, AECM에 의한 apoptois 유도 현상이 유의적으로 감소되어 AECM에 의한 Hep3B 세포의 apoptosis 유발에 caspase-3이 중요한 역할을 하고 있음을 알 수 있었다. 본 연구의 결과만으로 동충하초에 의한 간암세포의 증식억제 기전을 명확하게 제시할 수는 없으나, 이상의 결과들은 동충하초의 생화학적 항암기전 해석을 이해하는데 중요한 기초자료로서 활용될 수 있을 것으로 생각된다.

Keywords

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