• 系統編號 RG9605-0157
• 計畫中文名稱 補益類中藥保護幹細胞及影響癌病患腫瘤細胞之免疫相關之功能基因體研究(2-1)
• 計畫英文名稱 --
• 主管機關 行政院衛生署 • 計畫編號 CCMP95-RD-204
• 執行機構 台北醫學大學細胞及分子生物研究所
• 本期期間 9506 ~ 9606
• 報告頁數 24 頁 • 使用語言 中文
• 研究人員 施子弼；陳清祥；；；； Shih,Daniel TB；Chen,Chin-shiang；；；；
• 中文關鍵字 人體間葉幹細胞；乳癌細胞；補益類中藥；功能性基因體藥理；；；；
• 英文關鍵字 ；；；；；；；
• 中文摘要
研究目的: 探討補中益氣湯，降低放射線傷害副作用及其防護人體幹細胞之分子藥理機制。同時亦將平行比對分析補中益氣湯對乳癌組織細胞之影 響。分析比較上述檢定藥方作用於正常幹原細胞(例如:成體間質幹細胞)與腫瘤細胞(例如:乳癌初代細胞或細胞株)之細胞分子生理及其功能性基因體 表達層面之影響。 研究方法: 利用 RT-PCR 及定量 PCR 陣列等技術，分析細胞間交互作用分子及受體等蛋白表達之功能基因體變化影響。探討致癌 基因、腫瘤抑制基因、細胞凋亡基因、細胞週期基因、前驅細胞早期基因及生長分化訊息傳導基因之表達特性變化。利用螢光免疫染分析乳癌腫瘤細胞 附著分子，以及表面蛋白標記受放射線壓力及補中益氣湯藥效之影響。同時也利用細胞激素蛋白微陣列分析探討補中益氣湯影響前發炎因子、細胞 激素、趨化激素、介白素及其他細胞生長分化因子等蛋白表達之分析。結果與討論: 本計畫執行成果已完成平行比較分析正常體幹細胞及乳癌衍生細 胞，對補中益氣湯處理後的細胞及分子級藥理反應。其中包括放射線壓力下的細胞生長狀況，研究發現補中益氣湯對正常細胞的生長有正面修復或 保護效應，反之對乳癌衍生細胞則為負面效應。分析所表達的基因則可發現補中益氣湯可促進乳癌衍生細胞高量表現腫瘤抑制及移轉抑制基因，同 時也會抑制乳癌衍生細胞表現血管生成相關基因。分析其激素蛋白表達差異性時，也可發現補中益氣湯會影響乳癌組織衍生初代細胞對周邊其他組 織細胞的調控效用。本計畫之研究成果，可對中藥在個體幹細胞及腫瘤惡性細胞的藥理調節上有更完整的資訊，提供補益類中藥應用於癌症醫療輔 助效益的評估範例，並提供中藥應用於生理維護與組織修復機制及效用提供更有效之論證。建立一現代化中藥藥理鑑定的新方向，以助中藥之國際 化發展。
• 英文摘要 Aim: In this proposal study, we proposed to pursue a pharmacogenomic study on Bu-Yi prescription as a radiation therapy suppliment candidate for breast cancer patients. The aim of this study is to analyze the possible distinct pharmagenomic mechanism of selected Bu-Yi prescription on human normal stem cells and breast cancer cells under the influence of gamm-ray irradiation stress. Method: The drug effect on proliferation and stress/toxicity response in the primary
culture of mesenchymal stem/progenitor cells and cancer cells were qualitatively and quantitatively compared at the molecular basis. Specifically, Gene expressions of oncogenes, tumor suppressor genes, apoptotic genes, cell cycle genes, early genes and Notch signal pathway genes were compared by RT-PCR analysis. Stress/toxicity pathway response genes were compared by a Real-time Q-PCR array study. Cell adhersion/interaction molecules and cancer/stem cell markers were analyzed by immunoflouence stain. Breast cancer cell secreted factors (such as proinflammatory factor, cytokines and growth factors) were analyzed by cytokine protein array. Results & Discussion: In this study, we have simultaneously compared the cellular and molecular pharmagenetic responses to the Bu-Yi prescription between human mesenchymal stem cells and breast cancerous cells。Under the irradiation stress, our experimental results indicated that the Bu-Yi prescription exerts both protective and repair effect on the proliferation pontency of normal MSCs, while irradiation-damage effect of breast cancerous cells was augment by the Bu-Yi prescription. Consistantly, Tumor suppressor gene Pinin, and metastasis suppressor BRMS1 were indeed largly increased in breast cancerous cells after treated with the Bu-Yi prescription. In addition, we further found the Bu-Yi prescription diminished expression of angiogenic factors (such as VEGFR3 and VEGF-A) in breast cancerous cells. Take together, our results present better understanding distinct
pharmacogenomic effects of the Bu-Yi prescription on normal stem cell and cancerous cells. Our study also provides better insights on the benefits of the Chinese prescription supplement for cancer patient’s irradiation therapy at molecular level. Our experimental data exemplify the benefit of traditional Chinese prescriptions as therapeutic supplements. This study was supported by NSC national research program (CCMP95-RD-204).
