蜂肽誘導人類乳癌細胞株MCF-7凋亡之研究; Bee venom induce human breast cancer cell line MCF-7 apoptosis

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题名:	蜂肽誘導人類乳癌細胞株MCF-7凋亡之研究;Bee venom induce human breast cancer cell line MCF-7 apoptosis
作者:	廖士欣;Shih-Shin Liao
贡献者:	中國醫藥大學：營養學系碩士班
关键词:	蜂肽;細胞凋亡;粒腺體膜電位;活性氧成分;Bee venom;Apoptosis;Mitochondrial membrane potential;Reactive oxygen species
日期:	2007-07-13
上传时间:	2009-08-11 15:30:56 (UTC+8)
摘要:	Apis Mellifera品種的蜂肽(Bee venom)是一種具有多種藥理與生物活性的複合物質。在一些研究中証實，蜂肽可以減輕疼痛減少過敏反應。蜂肽已經被證實，可以抑制一些癌細胞增生與誘導凋亡，然而蜂肽在誘導乳癌細胞凋亡的機制清楚。在本研究發現，Apis Mellifera品種的蜂肽能抑制乳癌細胞株(MCF -7)生長，具有濃度與時間效應。透過流式細胞儀的分析，Apis Mellifera品種的蜂肽在1小時就會使MCF-7活性氧成份(Reactive oxygen species)釋出，並降低粒線體膜電位。經由DAPI染色法證實，Apis Mellifera品種的蜂肽可以會造成MCF-7的細胞凋亡，經由彗星試驗證實，Apis Mellifera品種的蜂肽可以會造成MCF-7的DNA損傷。經由西方墨點證實，Apis Mellifera品種的蜂肽可以活化Bax而抑制Bcl-2、Bcl-xL的表現，使粒腺體膜電位下降，從粒腺體釋放出cytochrome c，活化caspase-9並與Apaf-1結合，進而活化caspase-3 ，使MCF-7產生凋亡；Apis Mellifera品種的蜂肽也會增加Fas的表現而活化caspase-8，進而活化Bid，促使caspase-3活化。 Bee venom(BV) of Apis Melliferais(A.M.) an complex compound with pharmacology and biological activities. In some study has been confirmed that BV can alleviate pain and reduce allergy. Although, BV has been confirmed that inhibit proliferation and induce apoptosis in some cancer cell. However, the molecular mechanisms involved in BV-induced apoptosis are still unclear in human breast cancer cells. In this study, we find that BV inhibited proliferation of human breast cancer cell line (MCF-7) in the concentration-dependent and dose-dependent. The result of Flow cytometry demonstrated that BV decreased mitochondrial membrane potential(MMP) by releasing reactive oxygen species(ROS) in one hour. Apoptosis was also confirmed by DAPI staining of MCF-7 cells after exposed to different time of BV. Based on the results from Comet assay, it was also indicated that BV induced DNA damage in MCF-7 cells. Based on the result from western blotting, BV can reduce expression of Bcl-2 and Bcl-xL through Bax activation, and reduced MMP lead cytochrome c releasing from mitochondria, increase expression of caspase 9 , Apaf-1 and caspase 3. BV also increase expression of Fas , caspase 8 and tBid.
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