- 學位論文
桑葚水萃物輔助膀胱癌放化療分子機制及應用之研究
The study of molecular mechanism and application of chemotherapy and IR via mulberry water extract assistance
摘要
膀胱癌是泌尿系統第二好發的癌症，目前是台灣十大癌症中排行第七名；膀胱癌棘手的地方在於高復發性，通常罹患膀胱癌的患者有高達70%會再復發，復發之後轉移機率也會相對提高，此時死亡率也會增加。現今膀胱癌治療方針為根治性膀胱全切除手術(radical cystectomy)，並且利用泌尿道重建來取代原本膀胱的功能，但是這種治療方法必須終身攜帶尿袋，所以治療上開始採取局部切除膀胱並且配合化學藥物及放射線的治療。但不論化學藥物或是放射線在生理上都會對患者造成副作用，另外癌細胞也容易對化學藥物或是放射線產生抗藥性或是抗敏性導致治療失敗。為了解決當前對於膀胱癌治療上的盲點，目前正積極研究在治療上添加天然中草藥或食品作為輔助者(auxiliary)，幫助降低現階段臨床治療劑量來毒殺癌細胞，幫助病患降低生理上的副作用及抗藥性，甚至避免復發及轉移的發生。在本篇論文研究證實，桑葚水萃物(Mulberry water extract, MWE: 250-1500 ug/ml)對於膀胱癌細胞(TSGH 8301 human bladder cancer cells line)合併太平洋紫杉醇(paclitaxel, dose: 3 nM)或放射線(IR, dose: 10 Gy)都比單獨使用藥物有較佳的毒殺效果，並且更進一步發現桑葚萃取物在合併紫杉醇48小時或放射線24小時後都會造成TSGH 8301停滯在細胞週期中的G2/M phase，並且都表現出mitotic catastrophe會發生的多核現象(multinucleation)。在探討桑葚合併紫杉醇作用機轉部分，結果發現合併處理可以透過誘導PTEN磷酸化表現增加而抑制early endosome的生成，使細胞質膜無法正常分裂(cytokinesis)而導致癌細胞發生mitotic catastroph。另外在桑葚合併放射線部分，發現合併處理可以透過Ras/Raf/MEK/ERK/MAP kinase pathway來抑制Aur- B的磷酸化表現導致細胞無法正常分裂。 在動物實驗方面，利用xenograft model可以觀察到桑葚水萃物合併化療或是放射線上可以有效抑制腫瘤生長，另外利用組織切片染色觀察蛋白表現，結果可以與in vitro相符，證明桑葚水萃物合併化學藥物或是放射線可以透過不同途徑誘導癌細胞走向mitotic catastrophe。期望本論文結果可以應用於膀胱癌臨床輔助治療。
並列摘要
Bladder cancer is the second predilection urinary system cancer, cancer is currently Taiwan's top ten ranked seventh. The treatment of bladder cancer by cystectomy combined with the treatment of chemotherapy and radiation because of the prevention of cancer is not clean. But whether chemicals or radiation will cause the patient in the physical side effects, in addition to the cancer cells also easy to become resistant to chemotherapy or radiation, or allergy, leading to treatment failure. In order to solve the blind spot for the treatment of bladder cancer, and is now actively adding natural herbs or food as a sensitizer in the treatment, helping to reduce the dose of poison clinical stage cancer patients to help reduce the physiological Side effects and drug resistance, or even prevent recurrence and metastasis. Our studies found that Mulberry water extract (MWE: 250-1500 ug/ml) combined with paclitaxel (dose: 3 nM) or radiation (dose: 10 Gy) for bladder cancer cells line (TSGH 8301) than the use of drugs alone have a better cytotoxic effect, and further found that MWE combined with paclitaxel (MWE/paclitaxel) or IR (MWE/IR) will result cycle arrest in G2/M phase in 48 hours in TSGH 8301 cells line, and promotes mitotic catastrophe -related proteins, such as Aur- A, Plk1, Rho A and Cep55 expression. MWE/paclitaxel decreased the immunofluorescence stain of early endosome antigen 1 (EEA1) and increased the expression and phosphorylation of phosphatase and tensin homolog (PTEN) indicating that the regimen inhibited the formation of recycling endosome which is required for cytokinesis. MWE/IR decreased phospho-Aur- B expression via Ras/Raf/MEK/ERK/MAP kinase pathway to inhibit cytokinesis. We found further that two combined treatments induced apoptosis of cancer cells and caused cell death after 72 hours finally. In a TSGH 8301 xenograft model, MWE/Paclitaxel retarded tumor growth via activation of PTEN and Caspase 3 and MWE/IR via inhibited Aur- B. These data demonstrated a synergistic effect on the anticancer efficacy of paclitaxel or IR by MWE supplement via promoting mitotic catastrophe through activation of PTEN and inhibition of Aur- B, and provide a novel and effective therapeutic option for bladder cancer treatment strategies.
參考文獻
延伸閱讀
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