傳統中藥方劑對人類乳癌細胞增生抑制作用和細胞程式死亡機制之探討類

林俊清
高雄醫學大學藥學系
在全世界，每年幾乎增加一百萬個乳癌的新病例，在發生於女性的癌症病例中，乳癌佔了18%。以英國為例，每年約有14萬人死於乳癌。台灣地區由於經濟快速成長，飲食日漸西化，導致女性乳癌發生率急遽上升，目前乳癌已是僅次於子宮頸癌，排名女性第二好發的癌症。本研究目標乃從生命科學發展迅速的分子生物學理論與傳統中藥方劑之作用相互配合，深入地探索其抗乳癌之活性機轉，如此不僅可增加西方醫學對中醫藥作用的接受度，並賦與中醫藥複方藥效的現代化意義。經過初步篩選結果中發現傳統中藥方劑之癰瘍劑如散腫潰堅湯和紫草根牡蠣湯等在濃度300 μg/ml的濃度可達最好的增生抑制效果，散腫潰堅湯：88.6% (MCF-7)和82.7% (MDA-MB-231)；紫草根牡蠣湯：78.3% (MCF-7)和72.1% (MDA-MB-231) 生長抑制作用。因此接續以這二種方劑針對受測細胞株p53正常、ER表現的MCF-7和p53突變、ER不表現的 MDA-MB-231 cell line，其各個分子在細胞中含量的變化以釐清受測藥物的抗癌機制是透過p53-dependent或p53-independent的途徑，同時可清楚的建立出其抗癌機轉。整體研究分成三大步驟，第一是初步抗癌活性的建立，確認抑制癌細胞增生所需之有效抑制濃度，分別確認藥物濃度和作用時間與其效應的關係性，並初步地探討抑制癌細胞增生的狀況，包括細胞凋亡及細胞週期停止等等。第二則針對藥物對腫瘤細胞抑制其增生和誘發細胞凋亡之機轉確立，評估該純化成分對p53、p21、p27、cyclins, cdks、Fas/Fas ligand、MAPK (JNK, p38,ERK1/2)和Bcl-2 family的影響，並藉由專一分子的抑制劑或gene-based knockdown的方式來確認這些受影響的分子所扮演的角色，建立其完整的抗乳癌途徑。最後則以nude mice (in vivo)的方式確立藥物在in vivo活性的評估。
關鍵字：乳癌；散腫潰堅湯；紫草根牡蠣湯；細胞凋亡

The proliferative inhibition and apoptotic mechanism of Traditional Chinese Medicinal Prescription (TCMP) in human breast cancer Cells

Chun-Ching Lin
Graduate Institute of Natural Products, College of Pharmacy, Kaohsiung Medical University
About a million women in the world are found to have invasive breast cancer every year. Breast cancer, counted up 18 percent, is one of the leading cancers in women. In England, one hundred and forty thousands of women die from breast cancer. The breast cancer case increases rapidly in Taiwan. At present, breast cancer and cervical cancer, respectively, are the first and second leading cancers in the women of Taiwan. Our laboratory focuses on the traditional Chinese medicinal prescription (TCMP) with molecular biology to further study the active mechanism their anti-breast cancer activity. This allows a better appreciation of Chinese traditional medicines in the modern era and also a better understanding of its underlying potential for therapy, thus increasing its acceptance in Western medicine. From preliminary research and screening, San Zhong Kui Jian Tangand Zi Cao Gen Mu Li Tang had inhibitory effect on breast cancer cells proliferation. The TCMPs are therefore selected to test on two human breast cancer cell lines, MCF-7 and MDA-MB-231. Using different dosages of TCMP, our study investigates whether their inhibition on the breast cancer cells are through a p53-dependent or -independent pathway, in order to clarify the underlying mechanism of the pure TCMP anti-breast cancer activity.
關鍵字：p53；p21；Fas receptor；Fas ligand；Bcl-family protein；MAKP