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| 題 名 | 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(全程總報告)=A Functional Genomic Study of Some Bu-Yi Chinese Herbs' Pharmacologic Effects on Protection of Tissue Stem/ Progenitor Cells and Immuno-modulation of Patient's Cancer Cells (Final Report) |
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| 作 者 | 施子弼; | 書刊名 | 中醫藥年報 |
| 卷 期 | 28:7 2010.09[民99.09] |
| 頁 次 | 頁33-86 |
| 分類號 | 414.5 |
| 關鍵詞 | 人體間葉幹細胞; 乳癌細胞; 前列癌細胞; 補益類中藥; 功能性基因體藥理; Human mesenchymal stem cells; Breast cancer cells; Prostate cancer cells; Bu-Yi prescription; Pharmacogenomic study; |
| 語 文 | 中文(Chinese) |
| 中文摘要 | 研究背景及目的: 延續之前探討補益類中藥具保護放射線及癌症化學醫療藥劑壓力傷害的 人體正常幹細胞效果分析,本計畫應用既知具正面防護人體幹細胞之補益中 藥效用資訊,探討補益中藥對受放射線醫療處理之癌細胞及正常幹細胞影響 之差異,尋找中藥輔助癌症醫療應用契機。二年計畫執行期間,分別針對女 性乳癌及男性前列腺癌為例,探討此補益中藥對癌症醫療輔佐應用之可能影 響。第一年集中於探討補中益氣湯對乳癌組織細胞在物理化學傷害壓力下之 影響;第二年探討四物湯(4D)保護人體正常幹細胞作用與對男性前列腺癌癌 組織細胞之不同反應中,尋找中藥輔助癌症醫療應用契機。 研究方法: 應用細胞體外初代培養技術觀察細胞存亡或增殖以及其細胞表面標誌及 受體等蛋白表達之表型變化。並用RT-PCR、分子微陣列分析評估藥物作用於 癌細胞影響其基因表達探討致癌基因、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。 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癌衍生細胞,對 補益中藥處理後的細胞及分子級藥理反應發現,補中益氣湯對正常細胞的生 長有正面修復或保護效應,反之部份對乳癌衍生細胞則為負面效應。進一步 分析其基因表達的變化,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。其對於乳癌細胞可降低BRCA-1及BRCA-2之表現並明顯減少帶有AC133 之細胞表形,可增加抑制細胞週期之P27及細胞凋亡基因P53表現。除此之外,在針對腫瘤轉移基因進行array分析,癌細胞抑制基因PNN及乳癌轉移抑 制基因BRMS1其表現量,故其於輔助性治療上可能有正面之輔助放射線治療 效用。相對性的觀察補益四物湯劑對不同前列腺癌細胞在RI(放射線)傷害壓 力下顯現著不同之反應及效益。在AC133之cell surface marker之分析上,在病 患檢體細胞上,則未見相同之影響作用。在基因表現的分析上,補益四物湯 劑則抑制PSCA、P27之表現並促進細胞CXCR4表形增加,但其對不同癌細胞 表形及細胞株作用亦有所相異。病人細胞檢體研究結果未有一致性的影響。 進一步分析發現四物湯在RI(放射線)傷害壓力下對Androgenic dependent之 PCC2前列癌細胞有明顯的失去修復之加層效益,但對Androgenic independent 之前列癌細胞有明確抑制癌細胞之凋亡及促進週期基因Cyclind D等基因表之 負面作用,顯示四物湯對個人癌細胞輔助醫療前分子檢測之必要,補益四物湯劑對於前列腺癌之輔助性治療應用上,仍有待探討。除外,我們對六味丸中藥之胞輔助醫可能影響亦作了初步的探討,結果發現六味丸對前列腺癌腫 瘤病患細胞在RI(放射線)傷害壓力下亦有增加腫瘤基因PSCA及癌細胞之表面 附著相關因子CD44表型之表現等多項負面作用,但在沒RI傷害壓力情況下反 之對PCC3前列癌細胞有顯著的存活成長抑制效用與對正常幹細胞及未確定 癌之前列腺腫病患之保護效益有顯著差異,顯示六味丸之服用效用在於保護 正常細胞、防護癌之形成作用但不適用於放射醫療癌症效益,以及病症不同 用藥之效果具大差異。四物湯與六味丸均顯示著補益中藥在各個病患細胞保 護或修復之效用有顯著之差異。補益類對癌細胞之效用評估亦顯示著癌腫瘤 細胞表型多樣性與其對藥物反之個例特異性錯綜複雜,本計畫之研究結果顯 示應用細胞初代培養技術及功能基因體評估藥物影響細胞表形及其生理變化 之重要性。因其可 提供中藥應用於生理維護與組織修復機制及效用有效之論證並在分子藥理解析上可提供個病案例對中藥在個體幹細胞及腫瘤惡性細胞的個病案藥理較完整的資訊,唯各病患細胞表型之多樣性又礙於檢測病例數 之有限未能足夠檢體建構補益類中藥應用於癌症醫療輔助之通用效益藥理評估。 |
| 英文摘要 | Aim: In this proposal study, we proposed to pursue a pharmacogenomic study on Bu-Yi prescription (補中益氣湯) as a radiation therapeutic adjuvant for breast cancer and Su-Wu Tang (四物湯) for prostate cancer patient cells. The aim of this study is to analyze the distinct pharmagenomic influences of selected Bu-Yi drugs among human normal stem cells, breast cancer and prostate cancer cells under the influence of gamm-ray irradiation stress. Method: The molecular pharmacologic effect on the co-cultured cell 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. Cancer cell secreted factors (such as proinflammatory factor, cytokines and growth factors) were analyzed by cytokine protein array. Results and Discussion: In this study, we have compared the cellular and molecular pharmacologi- cal influences of the selective Bu-Yi drugs in human mesenchymal stem cells and breast or prostate cancerous cells, under the γ-ray irradiation stress. Most Bu-Yi drugs tested exerts both protective and repair effect to the normal MSCs, including the cell function. In contrast, the irradiation-damage effect of breast cancer cells was found augment by the Bu-Yi prescription (補中益氣湯). Consistantely, tumor sup-pressor Pinin, and metastasis suppressor BRMS1 genes were found largly increased in breast cancerous cells after the Bu-Yi prescription treatment, under the radiation stress condition. Interestingly, the Bu-Yi prescription was also found diminished ex- pressions of angiogenic factors (such as VEGFR3 and VEGF-A) in breast cancerous cells. Take together, our results presented distinct pharmacogenomic influences of the Bu-Yi prescription on normal stem cell and cancerous cells. This result provides better insights on the potential benefits of the Chinese prescription supplement for breast cancer patient’s irradiation therapy. On the prostate cancer drug study, however, our results showed that Su-Wu Tang (四物湯) enhances cell cycle gene Cyclin D expression, and inhibits cell apoptotic genes of prostate cancer patient tumor cells studied under the irradiation stress condition, indicating the negative effect may in- fluent the cancer irradiation therapy. The result also showed that Su-Wu Tang (四 物湯) exhibits heterogeneous responses to different prostate cancer pataient cells, prostate cancer cell lines and normal MSCs. This phenomenon is also shown in the Lio-Wei Wan (六味丸) study in which it exhibits enhance PSCA gene expression and CD44 cancer cell adhesion molecule of prostate cancer clls or cell lines, but on the other hand presented more preventive and protective effects to the non-cancer detected prostate patient and normal stem cells. Current study exemplify the benefit of traditional Chinese prescriptions as therapeutic supplements in some cases can be achieved by cell molecular biological analyses. It also warrant that the pharmacological effect is case sensitive and cancer cell phenotype dependent. By analyzed the Bu-Yi Chinese Drug’s harmacologic effect on protection and repair of tissue stem/progenitor during cancer patient tumor cells may benifit for personalized medication in use of Chinese Bu-yi drug as therapeutic adjuvant. This study was supported by NSC national research program (CCMP95-RD-204). |
本系統中英文摘要資訊取自各篇刊載內容。