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標題: 中藥當歸補血湯對大腸癌之影響
The Influence of the Chinese medicine Danggui Buxue Decoction on colon cancer
作者: 賴俊廷
Lai, Chun-Ting
關鍵字: Danggui Buxue Decoction;當歸補血湯.大腸癌;colon cancer;apoptosis;細胞凋亡
出版社: 生命科學系
大腸直腸癌是全球盛行率第三,死亡率第四的癌症。目前早期發現的大腸直腸癌治療以手術為主,而末期大腸癌則以化學治療為主,但癌症患者在接受化療之後,可能出現身體虛弱,免疫力下降的情形。許多病患會在化療時或化療之後服用中藥進補。其中當歸補血湯是一常用之中藥複方,由黃耆與當歸按5:1 的比例組成。近年來研究發現當歸補血湯可促進紅血球生成、調節免疫功能、抑制癌細胞生長等功能。本篇論文以當歸補血湯處理大腸癌細胞株HCT-8 及CT-26,觀察細胞的變化,並以CT-26植入Balb/c小鼠皮下建立大腸癌動物模式,來觀察當歸補血湯對腫瘤和實驗動物的影響。
在細胞實驗方面,經過細胞增殖分析法(MTT assay)分析後,發現細胞生長及存活率都會隨著藥物濃度和處理時間的增加而受到抑制,進ㄧ步以流式細胞儀(flow cytometry)分析細胞週期變化,發現當歸補血湯造成大腸癌細胞的細胞週期停滯於G0/G1期(G0/G1 arrest),而sub-G1 phase的百分比會隨著藥物濃度和處理時間的增加而升高。接著以Annexin V-FITC/PI stain、TUNEL分析後,確定當歸補血湯會誘導大腸癌細胞進行凋亡(apoptosis)。為了進ㄧ步了解當歸補血湯中哪些成分造成上述結果,我們取當歸補血湯中的活性成分ferulic acid、formononetin、Astragaloside IV處理大腸癌細胞株CT-26,發現當三種純物質濃度為10-4 M時,對CT-26生長的抑制率達10-25%,而將三種純物質合併處理後,其抑制率可達46%;以流式細胞儀分析細胞週期變化,發現三種純物質都會造成G0/G1 arrest,而sub-G1 phase的百分比會隨著藥物濃度和處理時間的增加而升高。

Colorectal cancer is the third most common cancer and the fourth most frequent cause of cancer deaths worldwide. The primary treatment of colon cancer is the surgical resection, and chemotherapy treat late stage colorectal cancer. The cancer patients may suffer from decrease of immunity after chemotherapy. A lot of cancer patients will take Chinese herb during or after chemotherapy. In recent years, Danggui Buxue Tang (DGBXT) has been proven to promote the production of red blood cell, to enhance the immune function and to inhibit tumor growth.
In this study, we try to investigate the anti-cancer activity of DGBXT on colon cancer cell line, CT-26 and HCT-8. We also try to investigate the influence of the decoction on colon cancer animal model. Our data show that dose and time dependent effects of DGBXT inhibited colon cancer cell line proliferation. We analyzed the cell population during cell cycle by flow cytometry, and demonstrated that DGBXT triggered the arrest of cell cycle in G0/G1 phase. Moreover DGBXT induced apoptosis in colorectal cell line CT-26 that measured by TUNEL staining and Annexin V-FITC/PI stain.
Cell viability and cell population analysis performed on CT-26 cell line in comparison to three known compounds in DGBXT - ferulic acid, formononetin and Astragaloside IV. Cell viability showed the inhibition rates of each individual compound was within 15%-25% at the concentration of 10-4M. The inhibition rate of the complex contained these above compounds was 46%. Cell population analysis demonstrated that ferulic acid, formononetin and Astragaloside IV triggered the arrest of cell cycle in G0/G1 phase and increased percentage of sub-G1 phase dose and time dependent.
The Balb/C mice bearing CT-26 cell line was used as animal model to study the effect of DGBXT on colon cancer. All mice divided into 4 groups as Group I: Feeding normal saline as control group; Group II: Feeding DGBXT; Group III: injecting 5-FU; Group IV: Feeding DGBXT and injecting 5-FU. Experiment results demonstrated that DGBXT significant inhibited tumor growth. Compared the 5-FU treatment with 5-FU combined DGBXT treatment, the toxicity and side effects such as leukopenia and weight loss which caused by 5-FU treatment were reduced by DGBXT. There were no significant changes in body weight, hepatic and renal functions in the group of mice fed DGBXT.
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