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멜라틴이 세포자멸사 유발에 의해 DU-145 세포증식에 미치는 영향

Melittin Inhibits DU-145 Cell Proliferation Through Induction of Apoptosis

大韓鍼灸學會誌= The journal of Korean Acupuncture & Moxibustion Society, v.26 no.3, 2009년, pp.49 - 58  

심윤섭 (경원대학교 한의과대학 침구학교실) ,  송호섭 (경원대학교 한의과대학 침구학교실)

초록
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목적 : 본 연구는 봉약침의 주요성분인 멜리틴이 전립선 암세포주인 DU-145 세포성장에 어떤 영향을 미치는지를 알아보기 위하여 시행하였다. 방법 : 멜리틴이 DU-145의 성장에 미치는 영향을 알아보기 위한 cell viability 측정으로는 WST-1 assay를, 세포자멸사의 관찰에는 DAPI(4,6-diamidino-2-phenylindole)와 TUNEL staining assay를 시행하였으며, 세포자멸사 조절단백질(calpain, Bax, caspase-3, -9, cleaved caspase-3, cleavaged PARP, cleaved caspase-9, Bcl-2, XIAP, cIAP2, Akt, p-Akt, MMP-2, MMP-13)의 관찰을 위하여 western blot analysis를 시행하였다. 결과 : 1. DU-145 세포에서 멜리틴을 처리한 후 세포자멸사가 유도되어 세포성장이 억제되었다. 2. 세포자멸사 관련 단백질 중 분리된 caspase-3, caspase-9은 유의한 증가를, Bcl-2, p-Akt, XIAP, cXIAP는 유의한 감소를 나타내었다. 결론 : 이상의 결과는 멜리틴이 인간 전립선암세포주인 DU-145의 세포자멸사를 유발함으로써 증식억제 효과가 있음을 나타낸 것으로, 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 도움이 될 것으로 기대된다.

주제어

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제안 방법

  • To delineate whether the inhibition of cell growth by the MT was due to increase of the induction of apoptosis, I evaluated change of the chromatin morphology of human prostate cancer c아Is using DAPI staining. Consistent with the loss of viability, apoptosis determined after 24ir treatment was increased in a dose dependent manner.

대상 데이터

  • The DU-145 human prostate cancer cell was obtained from ATCC(American Type Culture Collection, Rockville, MD). Prostate cells were cultured in RPMI-1640 medium(Life Technologies Inc.

데이터처리

  • Data were analyzed using one-way analysis of variance followed by Tuckey test as a post-hoc test. Differences were considered significant at p< 0.

이론/모형

  • 2). I also evaluated DU-145 cell apoptosis by TUNEL assay. As shown in Fig.
  • 5您)for 24hr, the cells were washed twice with PBS and fixed by incubation in 4% para-formald산lyde in PBS for 1 h at room temperature. TUNEL assays were performed by using the in situ Cell Death Detection Kit(Roche Diagonostics GmbH, Mannheim, Germany) according to manufacturers instructions. Total number of cells in a given area was determined by using DAPI nuclear staining.
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