傳統中藥桂枝茯苓丸(GFW)作為替代膀胱癌化學治療藥物的探討

The Investigation of a Traditional Chinese Medicine, Guizhi Fuling Wan（GFW）as an Intravesical Therapeutic Agent for Urothelial Carcinorma of the Bladder

呂啓禎(Chi-Chen Lu)

指導教授 ： 陳永恩 博士 ； 徐政達 博士

國立中正大學/理學院/生命科學系分子生物研究所/博士(2016年)

摘要

膀胱癌是世界上第二常見的生殖泌尿系統癌症，由於它的術後復發率很高，因此常規的治療會在經尿道的刮除術(TUR)後，給予膀胱灌注化療藥或Bacillus Calmette Guerin (BCG)的治療，但這些膀胱灌注藥物會引起嚴重的血尿，甚至感染肺結核的副作用，造成患者很大的困擾。因此我們想從傳統中藥裡找一個處方其對人體的副作用是小的而且具有抑制膀胱癌生長的效果。桂枝茯苓丸(GFW)是一個大家熟知之傳統中藥複方，曾被報導可用於治療肝癌和子宮頸癌，因此我們探討桂枝茯苓丸是否也有抑制膀胱癌生長的作用。我們利用細胞生長與毒殺試驗來確認桂枝茯苓丸與現行膀胱灌注藥物mitomycin C、epirubicin 和 cisplatin同樣具有抑制人類膀胱癌生長的效果，但是其對正常人類膀胱上皮細胞的毒殺性是非常小的。並且藉由流式細胞儀分析桂枝茯苓丸能引起人類膀胱癌細胞的細胞週期休止進而造成細胞的凋亡。接著更進一步研究桂枝茯苓丸在活體老鼠膀胱癌模型中抑制膀胱癌的效用和評估其副作用，得到桂枝茯苓丸與mitomycin C和BCG同樣有抑制活體老鼠膀胱癌生長的效果。最後我們藉由流式細胞儀和西方墨點法來探討桂枝茯苓丸抑制膀胱癌生長的機轉。結果顯示桂枝茯苓丸能造成膀胱癌細胞內活性氧化物質含量增加使細胞產生氧化壓力死亡。這結果讓桂枝茯苓丸成為膀胱癌膀胱灌注藥物的一個好選擇。

關鍵字

桂枝茯苓丸 ； 膀胱癌 ； 膀胱灌注 ； Bacillus Calmette Guerin ； Mitomycin C ； 活性氧化物質

並列摘要

Urothelial carcinoma is the second most common malignancy of the genitourinary tract in the world. The patients of bladder cancer have high risk of recurrence and need the administration of supplemental intravesical chemotherapy or Bacillus Calmette Guerin (BCG) after transurethral resection (TUR). However, current therapeutic agents often cause severe side effects of hematuria and miliary pulmonary tuberculosis. Therefore, we intend to find a traditional Chinese Medicine as a substitute of chemotherapy with enhanced efficacy and minimal toxicity are urgently required for the treatment of urothelial carcinoma. Guizhi Fuling Wan (GFW) is a well-known traditional Chinese herbal formula which has been reported to inhibit the tumor cell growth of hepatocellular carcinoma and cervical cancer. Therefore, we investigate whether GFW also inhibit the growth of urothelial carcinoma. In this study, the CCK-8 assay demonstrated that GFW had an effect on growth inhibition of human bladder cancer cell lines was comparable to that of mitomycin C, epirubicin and cisplatin, but had mild toxicity on normal human bladder epithelial cells, and flow cytometry assay showed that the major cytotoxic effect of GFW was the induction of cell cycle arrest and apoptosis in bladder cancer cells. Subsequently, we carried out in vivo study to further confirm the inhibitory effect of GFW on bladder cancer using a mouse model, and evaluated its side effects as compared with mitomycin C and Bacillus Calmette Guerin (BCG). The results demonstrated that GFW therapy effectively inhibited orthotopic tumor growth as same as BCG and mitomycin C. Finally, we further investigated the mechanisms underlying GFW-induced cell cycle arrest and tumor cells death by flow cytometer and Western blot analysis. A clear reactive oxygen species (ROS) activated pathway in response to GFW-treatment to illustrate GFW as a good candidate for intravesical chemotherapy against bladder cancer was observed.

並列關鍵字

Guizhi Fuling Wan (GFW) ； Urothelial carcinoma ； Intravesical ； Bacillus Calmette Guerin (BCG) ； Mitomycin C ； Reactive oxygen species

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