抗黴菌藥物對小鼠L-5178Y淋巴腫瘤細胞生長的抑制作用:誘發細胞凋亡與細胞週期停滯

The Growth Inhibition of Antifungal Drugs in Murine L-5178Y Lymphoma Cell: Induced

簡振欽(Chen-Chin Chien)

指導教授 ： 劉正民

臺北醫學大學/醫學院/醫學檢驗生物技術學研究所/碩士(2002年)

摘要

一些研究中指出抗黴菌藥物Griseofulvin (GF)或ketoconazole (KT)能夠造成腫瘤細胞細胞週期產生G2/M arrest或G0/G1 arrest，並誘發腫瘤細胞發生apoptosis。在本實驗中，我們選用四種抗黴菌藥物，包括Griseofulvin (GF)、Nystatin (NT)、Amphotericin B (AmB)以及Clotrimazole (CLT)，探討它們對murine L-5178Y lymphoma生長的影響。我們使用trypan blue染色計算腫瘤細胞存活率、利用propidium iodide染色及流式細胞儀分析腫瘤細胞的apoptosis以及cell cycle的變化，然後進一步使用agarose gel electrophoresis的技術去偵測腫瘤細胞DNA ladder的情形，最後利用western blotting assay去觀察 Bax protein的表現。在藥物濃度方面，分別以GF (10到80 mM)，AmB (1 到25mM)。CLT (1到25mM)以及NT (1到30mM)等五種不同濃度。結果發現GF、AmB以及CLT都能有效的誘發腫瘤細胞產生apoptosis，造成腫瘤細胞的死亡，apoptosis與藥物濃度成正相關性。此外，也發現GF能使腫瘤細胞產生G2/M arrest；而CLT則可使腫瘤細胞出現G0/G1 arrest。而NT則沒有任何作用。經由實驗結果，證實抗黴菌藥物griseofulvin、amphotericin B以及clotrimazole都具有誘發腫瘤細胞株產生apoptosis的能力，或許可以成為一種新的腫瘤治療方法。

關鍵字

Griseofulvin ； Ketoconazole ； Amphotericin B ； Clotrimazole ； Nystatin ； Murine L-5178Y lymphoma ； Apoptosis

並列摘要

Some studies have demonstrated that cancer cell G2/M or G0/G1 cell cycle arrest and apoptosis were induced by antifungal drugs, griseofulvin (GF) or ketoconazole (KT). In this study, four kinds of antifungal drugs were chosen which include griseofulvin (GF), nystatin (NT), amphotericin B (AmB) and clotrimazole (CLT). To study the effects of these drugs on the growth of murine L-5178Y tumor cells, the percentage of survival tumor cells, cell apoptosis and cell cycle were observed. The former was measured by using trypan blue stain and the latter was analyzed by flow cytometry. In addition, DNA laddering was performed by using agarose gel electrophoresis and Bax protein were detected by western blot assay. In this study, different concentrations of each drug were dissolved in the culture medium. The doses of each drug were calculated in the effective range of GF (from 10mM to 80mM), AmB (from 1mM to 25mM), CLT (from 1mM to 25mM) and NT (from 1mM to 30mM). The results showed that GF、CLT and AmB revealed dose dependent relation on tumor cell apoptosis and viability. In addition, cell cycle arrest was induced by treating with GF or CLT, contrary, NT showed no effect on tumor cells. The results demonstrated that the antifungal drugs, griseofulvin, clotrimazole and amphotericin B may induce apoptosis in L-5178Y lymphoma cells and might to prove to be a novel therapy for lymphocytic cancer.

並列關鍵字

Griseofulvin ； Ketoconazole ； Amphotericin B ； Clotrimazole ； Nystatin ； Murine L-5178Y lymphoma ； Apoptosis

參考文獻

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