| Abstract: | 研究目的 中藥對癌症所引起的諸多症狀的治療能有所助益，已是不爭的事實。而以化學療法的觀點，發展並研究出中藥的有效成份，以供臨床上抗發炎、抗過敏及抗癌的應用，在近年來受到廣泛注意。目前已有多種中藥已被確認具有抗發炎，抗過敏及抗癌的活性，而其中有關活性的有效成份，部分已被單離並經分析鑑定。而中藥之抗癌有效成份之作用機轉，為有效地促使凋謝死亡及抑制癌瘤細胞增生，侵襲及轉移，但對其系統生物學方面的研究及建立尚很少。因此本計畫擬進行（1）分析57種中藥對癌瘤細胞如肺癌及肝癌等細胞之cell adhesion，motility，migration 及invasion活性的影響；（2）探討57種中藥對癌瘤細胞如肺癌及肝癌等細胞之ECM分解相關基因表現量的影響，包括 MMPs、TIMPs、t-PA、u-PA及PAIs；(3)利用動物模式探討中藥抑制癌瘤細胞的生長。研究方法本實驗利用了MTT assay探討中藥對肺癌細胞A549的細胞存活率；也利用Boyden chamber 分析中藥對肺癌細胞之cell adhesion、motility、migration 及invasion活性的影響。我們也利用gelatin zymograohy及western blotting分析中藥對肺癌細胞ECM分解有關基因表現量的影響。最後我們利用Lewis lung carcinoma及C57BL/6的動物模式探討中藥對抑制癌瘤細胞生長的影響。結果與討論我們取中藥萃取物，加入A549細胞株中，利用這些中藥萃取物在100？慊/ml是否影響A549的細胞生長能力、invasion及migration能力，我們將中藥分成三大類，第一大類是在100？慊/ml的濃度時，不影響細胞生長能力、卻會抑制細胞invasion及migration能力；第二大類是在100？慊/ml的濃度時，會抑制細胞生長能力、細胞invasion及migration能力；第三大類是在100？慊/ml的濃度時，完全不會抑制細胞生長能力、細胞invasion及migration能力。而我們發現紫丁地、卷柏、夏枯草、葉下珠、骨碎補及欖仁樹的葉這六種中藥萃取物抑制A549細胞株invasion及migration能力是經由抑制癌症轉移及侵襲的相關基因MMP-2及u-PA的表現及其抑制劑之間的表現所造成的。另外，我們也在mRNA level方面看到類似的抑制結果。而我們也在動物模式方面，更加應證了這些中藥萃取物會抑制腫瘤轉移及侵襲的能力。
Aim It has been well established that Chinese herbal medicines have beneficial effects for various symptoms of cancers. For chemotherapy, a great deal of attention has been focused on studies and researches for the active ingredients of Chinese herbal medicines for the treatment of inflammation, allergy, and cancers. So far, various Chinese herbal medicines have been confirmed to possess an activity against inflammation, allergy and cancers, while some of active ingredients have been isolated and identified. The anticancer mechanisms of these active ingredients are to effectively promote apoptosis, inhibit the proliferation, invasion and metastasis of cancer cells. However, there are still inadequate researches to establish the detailed mechanisms. (1) to analyze the impacts of 57 Chinese herbal medicines on the cell adhesion, motility, migration and invasion of various cancer cells, including cancers of lung and liver cancer (2) to investigate the impacts of 57 Chinese herbal medicines on the expression of genes relevant to ECM-degradation, such as MMPs, TIMPs, t-PA, u-PA, and PAIs of various cancer cells, including cancers of lung and liver cancer; and (3) to use the animal model to evaluate the impacts by the addition of Chinese herbal medicines and their applications. Method With a few established techniques, including gelatin, casein zymography, immunoblotting, RT-PCR and Boyden chamber for MMPs, u-PA, and t-PA, our Lab is very capable of performing studies on genes relevant to ECM-degradation, such as MMPs, TIMPs, plasmin, t-PA, and u-PA to clarify their involvements in anticancer ability.We also use the animal model incuding Lewis lung carcinoma and C57BL/6 miceto clarify the Chinese herbal medicines in anticancer ability. Results & Discussion We analyaze the impacts on the cell adhesion, motility, migration and invasion of A549 lung cancer cell using 57 Chinese herbal medicines .57 Chinese herbal medicines were divided into three groups based on the inhibitoy ability of viability, invasion and migration. With 6 Chinese herbal medicines including Phyllanthus urinaria , Selaginella tamariscina , Terminalia catappa , Davallia mariesii Moore , Prunella vulgaris and Syringa vulgaris treatment on A549 cell showed may decrease the expressions of MMP-2 and u-PA in a concentration- manner. Further analysis with semi-quantitative RT-PCR showed that these Chinese herbal medicines may regulate the expressions of MMP-2 and u-PA on the transcriptional level. Furthermore,use the animal model also showed that these Chinese herbal medicines may inhibit the tumor growth and migration in vivo. |
