三環藥物Teroxirone對非小細胞肺癌細胞增生機制的探討

Abstract

肺癌細胞依其組織型態可分為兩類:小細胞肺癌 (small cell lung cancer, SCLC) 以及非小細胞肺癌 (non-small cell lung cancer, NSCLC)，而其中大多數屬於非小細胞肺癌。目前在臨床治療上經常使用的抗癌藥物 (例如: camptothecin, doxorubicin, etoposide, cisplatin) 容易產生抗藥性，導致治療的困難，因此本文開發目前臨床上還未徹底了解，但具有抗癌效果的化合物。 研究目標是發展小分子且低劑量的三環氧化物teroxirone，了解teroxirone如何引發細胞凋亡。實驗以MTT及細胞群落分析 (colony formation assay) 確認teroxirone確實能降低非小細胞肺癌的存活率。接著要確認非小細胞肺癌細胞的存活率下降的確是由於細胞凋亡所造成。再利用單細胞凝膠電泳 (single cell gel electrophoresis (SCGE) / comet assay)，偵測teroxirone是否造成DNA 損傷。再利用propidium iodide (PI) 染色觀察teroxirone對細胞週期的影響，再利用二維流式細胞儀及DNA片段化實驗觀察teroxirone所誘導的細胞凋亡。 實驗結果顯示，teroxirone都會對NSCLC細胞造成不同程度DNA損傷，且細胞凋亡與p53的基因型有關。Lung cancer according to biological characteristics can be classified two types: small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC), most of which belong to non-small cell lung cancer. Clinical treatment used anticancer drugs (ex: camptothecin, doxorubicin, etoposide, cisplatin) frequently is easy to become resistant, leading to difficulties of treatment. The purpose of the thesis is to study the effectiveness of potential anticancer drugs.

The goal of this study is to understand how low doses of teroxirone induce NSCLC cell apoptosis. Teroxirone can reduce the proliferation rate of non-small cell lung cancer on indicated by MTT assay and colony forming assay. To confirm the reduced NSCLC cell growth rate is indeed due to cell death. To be have used the single cell gel electrophoresis (SCGE) / comet assay to detect DNA damage. To be have used propidium iodide (PI) staining to observe that impact of teroxirone on cell cycle and the induced apoptosis by flow cytometry. Double staining with PI and Annexin V – FITC of cell death for apoptosis. At the same time, DNA fragmentation also confirmed teroxirone indeed induced cells apoptosis.

The experimental results showed that teroxirone caused different degrees of DNA damage in NSCLC cells, but is caused apoptosis only occurred in p53 wild-type cells.