臺灣博碩士論文加值系統

相關研究指出Prostaglandin F2 Receptor Inhibitor (PTGFRN)，和人類部分腫瘤的生長與血管的新生有關。而多型性神經膠質母細胞瘤(Glioblastoma multiforme，簡稱GBM)，為最常見的原發性惡性腦瘤，病程惡化速度快、腫瘤浸潤性高、手術不易完全清除乾淨，使其成為神經膠質瘤(Glioma)中惡性程度最高的腫瘤，若只接受開刀治療中位存活時間(median survival time) 只有4個月，接受傳統放射治療中位存活時間可延長至9~10個月，而目前多型性膠質母細胞瘤的標準治療方式是同步藥物與放射治療，但中位存活時間也頂多延長至15個月，5年存活率僅有3~5%，預後仍不盡理想。因此想探討人類與GBM相關的基因，期望日後能以基因療法得到較好的治療效果。
本實驗利用即時聚合酶鏈鎖反應(Real-time PCR)、西方墨點法(Western blot)以及神經膠質瘤(Glioma)組織微陣列(tissue microarray)進行免疫組織染色(immunohistochemistry stain)，來評估PTGFRN在神經膠質瘤細胞之表現。以上實驗結果皆顯示PTGFRN在神經膠質瘤細胞組織的mRNA與蛋白質的表現量都比正常腦細胞組織來的高，而且其表現量的多寡與世界衛生組織(WHO)的腫瘤分級具有正相關性。未來的研究方向將透過抑制此基因，觀察腫瘤細胞生長速率、侵襲能力以及對細胞週期的影響，藉此驗證PTGFRN是否能成為未來治療神經膠質瘤的標的基因。

Recent studies have shown Prostaglandin F2 Receptor Inhibitor (PTGFRN) is related to angiogenesis and tumor overgrowth in human. De novo glioblastoma multiforme (GBM) is the most common and highly malignant form of brain tumor. It is aggressive and challenging to be treated. Due to their finger-like tentacles, they are difficult to be completely removed by surgery. Median survival time for patients receiving only surgical resection is 4 months. The median survival time is extended to 9 to 10 months with conventional radiation therapy. The current standard treatment for glioblastoma is concurrent chemo-radiation therapy (CCRT) with median survival time of 15 months. 5-year survival rate of GBM ≤ 5% which is less than ideal. The objective of this study is to explore human genes that is associated with GBM. We sincerely hope to achieve better outcomes via pharmacology target therapy in the future.
U87MG, LN229 and GBM8401 cell lines were cultured to determine the mRNA and protein expression level of PTGFRN in gliomas. For the immunohistochemistry (IHC) stain, 85 glioma brain tissues and 5 non-neoplastic brain tissues were collected to make the tissue microarray. The expression level of PTGFRN was evaluated by real-time PCR, western blot and IHC. Our results showed mRNA and protein expression level of PTGFRN is up-regulated in glioma cells/tissues as compared to normal brain tissues. The expression level of PTGFRN is thus positively correlated to WHO tumor grades of gliomas. PTGFRN serve as a promising candidate for future research to place upon in observing its inhibition on the effects on cell cycle, growth rate, and invasion ability. This may serve to verify in determining whether PTGFRN can act as new therapeutic target for glioma patients.

目錄 I
表目錄 IV
圖目錄 V
縮寫表 VI
中文摘要 VIII
Abstract IX
第一章 緒論 1
第一節 神經膠質瘤(Glioma) 1
第二節 與神經膠質瘤相關基因 2
第三節 Prostaglandin F2 Receptor Negative Regulator 3
第四節 研究動機與目的 4
第二章 材料與方法 5
第一節 實驗材料 5
壹、 細胞及組織材料 5
貳、 藥品試劑 5
參、 儀器設備 8
第二節 實驗方法 9
壹、 細胞培養(Cell Culture) 9
貳、 反轉錄聚合酶鏈鎖反應(RT-PCR) 10
參、 即時聚合酶鏈鎖反應(Real-time PCR) 12
肆、 西方墨點法(Western blot) 13
伍、 石蠟切片(Paraffin Block Section) 16
陸、 蘇木紫-伊紅染色(Hematoxylin & Eosin stain) 17
柒、 免疫組織化學染色(Immunohistochemistry stain) 18
捌、 數據分析 18
第三章 結果 19
第一節 GEO Profiles資料中Gliomas WHO分級與PTGFRN mRNA表現呈現正相關性 19
第二節 PTGFRN在神經膠質瘤細胞株有較高的mRNA表現 19
第三節 PTGFRN在神經膠質瘤細胞株中有過度蛋白質表現 19
第四節 Glioma病患腦組織中利用免疫組織化學染色法證實PTGFRN蛋白質表現量和世界衛生組織(WHO)腫瘤分級具有正相關性 20
第五節 Glioma腦腫瘤PTGFRN免疫組織化學染色結果與病患存活時間具負相關性 21
第四章 討論 22
第五章 結論 24
Tables (表) 25
Figures (圖) 27
第六章 參考資料 39

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