- 學位論文
探討廣藿香醇抑制血癌細胞株增生藉由誘導細胞凋亡及細胞週期G1期停滯之相關機轉調控
Proliferation Inhibition of Patchouli Alcohol (PA) in Human Leukemia Cells by Apoptosis Induction and Cell Cycle G1 Arrest
摘要
在本論文中，研究結果顯示廣藿香水萃取物 (WEPC) 可誘導人類白血病細胞株細胞凋亡，然而植物萃取物成分複雜度仍有待探討，因此，利用廣藿香內有效成分之一的廣藿香醇 (Patchouli Alcohol；PA)進一步探討有效成分對於人類血癌細胞株之影響。 首先在本碩士論文所使用的WEPC是中草藥植物廣藿香 (Pogostemon cablin；PC) 的水萃取物，在傳統上廣藿香常用來治療感冒、反胃、腹瀉、頭痛及發燒等症狀；其中一種有效成分為廣藿香醇，有研究指出廣藿香醇具有抑制大腸癌細胞 (HCT116、SW480) 細胞增生之抗腫瘤特性。然而，兩者藥物對於白血病治療上相關機制目前還不明確，因此在本論文中將會進一步探討廣藿香萃取物對於人類急性骨隨性白血病細胞株 U937 及人類慢性顆粒性細胞白血病細胞株 K562 的抗腫瘤生長之相關機制。 本研究目的是想了解廣藿香萃取物及有效成分在體外試驗中的抗腫瘤特性，藉由廣藿香萃取物刺激血癌細胞使細胞停止不正常增生，進而達到治療效果。透過流式細胞儀分析細胞週期變化、DNA含量分析。再藉由 PCR array 及 Real-time PCR分析細胞週期基因表現量，如：Cyclins/CDKs及細胞凋亡，如：Bcl-2。再進一步利用SDS-PAGE 和 Western blotting ，分析調控細胞週期及細胞凋亡之相關蛋白質含量。 除了抑制血癌細胞增殖和誘導細胞凋亡外，其他機制證明了廣藿香萃取物的抗血癌作用，如：Caspase-3 活性試驗、粒線體膜電位(ΔѰm ) 改變等。 近年來天然物質萃取之研究已經日益增加，目前有很多已經被證實有相當顯著的功效，然而本研究也證實廣藿香萃取物可能具有抗腫瘤的功效，並可推論廣藿香的藥理作用在於抗腫瘤上的應用將非常重要。
並列摘要
Pogostemon cablin (PC) is a plant found in traditional Chinese medicine, traditionally used to treat the common cold, nausea, diarrhea, headaches, and fevers. Water extract from Pogostemon cablin (WEPC) has also been shown to induce apoptosis in human leukemia cells. One of its active ingredients in PC is patchouli alcohol (PA), which has been shown to inhibit cell proliferation of colon cancer cell lines HCT116 and SW480. This study investigated the influence of patchouli alcohol (PA), on human leukemia cells. PA has been shown to possess neuroprotective, anti-influenza and anti-inflammatory activities; however, the mechanisms underlying these effects have yet to be elucidated. This study sought to identify the mechanisms of PC extracts with regard to the treatment of human acute myeloid leukemia (AML) cell line U937 and the chronic myelogenous leukemia (CML) cell line K562. We focused on the anti-tumor properties of WEPC and PA in vitro in halting abnormal cell proliferation. Cell cycle progression and DNA content were analyzed using flow cytometry. Gene expression of cyclins/CDKs and apoptosis in Bcl-2 cell cycles were analyzed using PCR arrays, and real-time PCR. SDS-PAGE and western blotting analysis were used to identify the proteins associated with various cell cycles and apoptosis. The results of caspase-3 activity and mitochondrial membrane potential (ΔѰm) demonstrate that WEPC and PA possess anticarcinogenic mechanisms other than the inhibition of cell proliferation and induced apoptosis. Our findings provide further evidence of the anti-tumor effects of WEPC and PA with potential pharmacological benefits for the treatment of cancer.