中草藥BP004誘導管腔A型乳腺癌細胞凋亡;BP004, a Traditional Chinese Medicine, induces apoptosis in human luminal A breast cancer cell lines

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题名:	中草藥BP004誘導管腔A型乳腺癌細胞凋亡;BP004, a Traditional Chinese Medicine, induces apoptosis in human luminal A breast cancer cell lines
作者:	林君潔;Lin,Jun-jie
贡献者:	系統生物與生物資訊研究所
关键词:	乳癌;蒲公英;中草藥;基底A型乳癌;細胞凋亡;Breast cancer;Dandelion;Traditional Chinese medicine;Luminal A;Apoptosis
日期:	2014-08-27
上传时间:	2014-10-15 14:45:15 (UTC+8)
出版者:	國立中央大學
摘要:	乳癌發生率及死亡率逐年攀升，但是主流醫學並沒有辦法延長患者生命。雖然多本中醫古籍記載具有清熱解毒、活血化瘀等特性的中藥能緩解癌症病徵，主流醫學還沒有對它們的療效做科學驗證。我們利用高通量細胞學藥物篩選系統從約60種此類中草藥篩選出蒲公英萃取物BP004為候選治療藥物，並利用多種量化方法證實BP004對乳癌細胞的確有抑制生長及毒殺的效果。首先我們使用細胞存活率試驗證明BP004能抑制基底A型乳癌細胞MCF-7細胞生長。其對基底A型乳癌細胞MCF-7的50% 生長抑制所需濃度(IC50)為550μg/ml; 對基底A型乳癌細胞T47D為700μg/ml。對三陰性乳癌細胞MDA-MB-231、her-2/neu陽性乳癌細胞MDA-MB-453及正常乳腺上皮細胞 H184B5F5/M10無生長抑制的反應。我們接著使用流式細胞儀觀察上列細胞株對BP004處理的反應，發現基底A型乳癌細胞MCF-7會發生細胞凋亡。並使用西方墨點法量化經不同濃度BP004處理後各細胞株蛋白質的表現量，並發現基底A型乳癌細胞MCF-7的BAX, CASP-7, FAS等蛋白質表現量在BP004濃度為200-400 μg/ml BCL-2時上升，其BCL-2 蛋白質則隨之下降，表示BP004能使細胞走向內部凋亡。最後我們透過全基因體表現量分析，證實BP004除了向上調節基底A型乳癌細胞MCF-7細胞凋亡。此外，我們還發現有細胞週期及細胞外來刺激反應等功能。以上結果顯示BP004有潛力成為基底A型乳癌細胞之乳癌新藥。未來我們會強化過去的研究外，也會繼續進行動物層次研究，對於BP004是否可當作抗癌藥物需要再作更深入的評估其抗癌的功效。我們希望藉由臨床樣本的數據，以高通量微陣列晶片的技術利用系統生物學來整合基因調控元素及蛋白質交互作用間複雜的關係為藥物BP004的治療建立出可鑑別的乳癌分子標記。;Breast cancer incidence and mortality rates haves increased over the years, but due to limitation of current medicines, there were no effective way extending the lives of patients. Although many Chinese ancient books had recorded the benefits of Chinese medicine in terms of cancer treatments by alleviating fever, de-toxication and, eliminating the blood flow by promoting blood circulation, the mainstream medicines still haves not yet approve scientifically their efficacies and use. In this study, we used high-throughput drug screening cytology system (High-throughput Drug Screening System Cytology, HT-DSSC) to screen 60 herbal extracts, and BP004 (The dandelion extract) was the candidate for our study. BP004 was found to inhibit the growth of luminal-A breast cancer cell line MCF-7 by inducing cell apoptosis; however, none of the triple-negative breast cancer cell lines such as MDA-MB 231, HER2-positive breast cancer cell lines MDA-MB-453 and normal mammary epithelial cell H184B5F5/M10 were effected. The extrinsic and intrinsic apoptosis pathways were examined by Western blot. We found that BAX, CASP-7, and FAS proteins were increased in expression while, BCL-2 was decreased expression, indicating cells undergo apoptosis through the intrinsic pathway. Finally, using microarray analysis, genes involved with "cell cycle" and "cellular response to stimulus were also identified". These results indicated that BP004 has is the potential adjuvant drug in the treatment of breast cancer cells. In the future, we will conduct animal experiments, to further confirm the effectiveness of BP004 in the treatment of breast cancer. We use of the technology for high-throughput microarray systems biology to integrate the complex relationships between elements of gene regulation and protein interactions for drug BP004 establish a molecular markers for breast cancer classification.
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