Chrysin抑制人類神經膠質母細胞瘤的轉移及增強Temozolomide作用機轉之研究

The study of chrysin on metastasis suppression and temozolomide sensitizing in human glioblastoma cells

謝東穎(Tung-Ying Hsieh)

指導教授 ： 林志立

中山醫學大學/醫學院/醫學研究所/碩士(2011年)

https://doi.org/10.6834/CSMU.2011.00049

摘要

神經膠細胞瘤Glioblastoma multiforme（GBM），是腦瘤中最常見的一種，也是腦瘤中致死率最高的一種。而目前針對GBM的治療的臨床主流用藥為Temozolomide（TMZ）；TMZ的優點是口服便能達到所需濃度，而缺點則是當O-6-甲基鳥嘌呤甲基轉移酶（MGMT）表現量上升時，TMZ所造成的DNA損傷便會被逆轉，使得腫瘤細胞不會走向死亡，有機率在距離原發位置3公分內復發，且20~30％的患者有產生遠端轉移的可能，而復發與轉移則是腦部腫瘤死亡的主要原因，因此我們希望能找到與其搭配，降低轉移、增加療效的藥物。
在過去文獻中指出松類萃取物中富含的生物類黃酮有抗癌的效果；因此我們取了幾樣台灣五葉松萃取物進行實驗，發現其中的白楊素（Chrysin）在傷口癒合實驗（Wound Healing Assay）與Boyden Chamber Assay對於腫瘤細胞的侵襲與轉移有顯著抑制的現象。因此我們想探討TMZ與Chrysin的合併治療對於GBM的治療或者是預後會不會有較大的改善。因此我們利用Western Blot方式觀察Chrysin影響腫瘤細胞走向凋亡的可能路徑，但結果卻發現腫瘤細胞並非走向細胞凋亡，反而是走向細胞自噬性死亡，並且是透過活化AMPK，抑制mTOR使得細胞走向自噬性死亡；除此之外透過觀察MGMT的表現量，發現加入Chrysin 合併處理過後，MGMT的表現量有下降的趨勢，表示加入Chrysin合併處理之後，不只能增強TMZ抑制腫瘤遷移與轉移的能力，並能改善TMZ治療後腫瘤容易復發的缺點。

關鍵字

多型性神經膠母細胞瘤 ； Temozolomide ； 自噬性細胞死亡 ； 台灣五葉松 ； O-6-甲基鳥嘌呤甲基轉移酶

並列摘要

Glioblastoma multiforme (GBM) is the most frequently malignant adult tumor, and the death rate is also the highest one. At present, Temozolomide (TMZ) is one of the standard chemotherapy agent to treat GBM, and TMZ has the advantage of oral administration will be able to achieve the required concentration, but the disadvantage is that when the activation of O-6-methyl-guanine methyltransferase (MGMT) , The DNA damage that TMZ caused would be reversed, the tumor cells will not to die, and patients have a chance at a distance of 3 cm within the primary site recurrence, and 20 ~30% of patients had the possibility of distant metastases. The recurrence and metastasis is the major cause of death of brain tumors, so we hope to find the match with TMZ, reducing the metastasis, and increase the efficacy of drugs.
Pine extracts containing rich in bioflavonoids. In present study, bioflavonoids showed the anti-cancer activity on cancer cells ; so we took a few extracts of Taiwan Pinus morrisonicola Hayata to experiments and the results showed that the Chrysin significantly inhibited the invasion and migration ability of GBM cells in wound healing assay and Boyden Chamber assay. Therefore, we would like to discuss the combined treatment of TMZ and Chrysin for the treatment or the prognosis of GBM will have a greater improvement. So we use Western Blot assay to observe the possible pathway that Chrysin cause the tumor cell into autophagic cell death instead of apoptosis, and found that Chrysin stimulating the activation of AMPK and attenuating of mTOR makes cells to autophagic cell death; in addition, through observe the expression of MGMT found that after combined treatment of Chrysin and TMZ, MGMT expression was decreased. Taken together, this results suggest that after the combined treatment of TMZ and Chrysin, not only enhanced the ability of inhibits tumor migration/invasion and the anti-cancer effect of TMZ treatment ,but also can improve the shortcomings of the tumor in recurrent.

並列關鍵字

Glioblastma multiforme ； Temozolomide ； Autophagic cell death ； Pinus morrisonicola Hayata ； O-6-methyl-guanine methyltransferase

參考文獻

[1] Quick,A, et al. Current therapeutic paradigms in glioblastoma.

Rev Recent Clin Trials,2010.5(1):p. 14-27.

[2] Aldape et al.Molecular epidemiology of glioblastoma.

Cancer J. 9 (2003), pp. 99–106.

[4] Benjamin et al. Classification of glioblastoma multiforme in adults by molecular genetics, Cancer J. 9 (2003), pp. 82–90.