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Effect of a Fibrinolytic Enzyme (BK-17) from Bacillus subtilis on Apoptosis Induction in AGS and T24 Human Carcinoma Cells

인간 암세포인 AGS와 T24에서의 apoptosis 유도에 미치는 Bacillus subtilis 혈전용해효소 BK-17의 영향

  • Baik, Hyun (Amaranth-Cosmetics Co.) ;
  • Seo, Min Jeong (Department of Biotechnology, Dong-A University) ;
  • Kim, Min Jeong (Department of Biotechnology, Dong-A University) ;
  • Lee, Hye Hyeon (Department of Biotechnology, Dong-A University) ;
  • Kang, Byoung Won (Medi-Farm Industrialization Research Center, Dong-A University) ;
  • Park, Jeong Uck (Medi-Farm Industrialization Research Center, Dong-A University) ;
  • Choi, Yung Hyun (Department of Biochemistry, College of Oriental Medicine and Research Institute of Oriental Medicine, Department of Biomaterial Control, Graduate School and Blue-Bio Industry Regional Innovation Center, Dongeui University) ;
  • Seo, Kwon Il (Department of Food Nutrition, Sunchon National University) ;
  • Jeong, Yong Kee (Department of Biotechnology, Dong-A University)
  • 백현 (아마라스화장품(주) 부산) ;
  • 서민정 (동아대학교 생명공학과) ;
  • 김민정 (동아대학교 생명공학과) ;
  • 이혜현 (동아대학교 생명공학과) ;
  • 강병원 (동아대학교 Medi-Farm 산업화 연구사업단) ;
  • 박정욱 (동아대학교 Medi-Farm 산업화 연구사업단) ;
  • 최영현 (동의대학교 한의학과) ;
  • 서권일 (순천대학교 식품영양학과) ;
  • 정영기 (동아대학교 생명공학과)
  • Received : 2013.06.18
  • Accepted : 2013.10.26
  • Published : 2013.10.30

Abstract

To investigate the effects of a fibrinolytic enzyme, BK-17, on the growth of human cancer cells, we performed various biochemical experiments, including cell proliferation and viability, and investigated subsequent morphological changes and apoptosis induction. BK-17 treatment of AGS human gastric and T24 human bladder carcinoma cells decreased the viability and the proliferation of the cells in a concentration-dependent manner. Microscopic studies indicated that the antiproliferative effects of the BK-17 treatment were associated with morphological changes, such as membrane shrinking, cell rounding up, and the formation of apoptotic bodies, indicating that BK-17 induced apoptosis in the cell lines. Of note, RT-PCR and Western blotting data indicated that the BK-17 treatment induced the down-regulation of antiapoptotic Bcl-2 members, Bcl-2 and $Bcl-X_L$, and the up-regulation of proapoptotic Bax members, Bax and Bad, in the AGS cells. BK-17-induced apoptosis of AGS cells was involved in the proteolytic activation of caspase-3, caspase-8, and caspase-9. Taken together, these findings suggest that BK-17 is associated with the induction of apoptotic cell death.

인간 암세포의 생육에 미치는 혈전용해효소(BK-1)의 영향을 조사하기 위해, 세포증식, 생존력, 형태변화 및 apoptosis 유도 등을 포함한 여러 가지 생화학적 실험을 하였다. 그 결과, AGS 인간 위장 암세포 및 T24 인간 방광 암세포상에의 BK-17 처리는 그 암세포들의 생존력 및 생율을 농도의존적 방법으로 감소시켰다. 현미경 관찰은, BK-17 처리에 의한 항 생육 효과는 막 수축, 세포의 rounding up, apoptotic bodies와 같은 형태학적 변화를 나타내었다. 특히, RT-PCR과 Western blotting data는, BK-17 처리가 항 apoptosis Bcl-2 군들 특히 Bcl-2, and $Bcl-X_L$의 down-regulation 그리고 AGS 세포에서, apoptosis 촉진 매개체 Bax와 Bad의 up-regulation를 유도했다는 것을 보여주었다. BK-17에 의해 유도된 AGS 세포의 apoptosis는 caspase-3, caspase-8 그리고 caspase-9의 단백질가수분해 활성과 관련이 있었다. 이상의 결과를 볼 때, BK-17은 apoptotic cell death의 유도와 밀접한 관련이 있다는 것을 보여주고 있다.

Keywords

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