我們藉由西方點墨法（Western blotting）和免疫組織染色法（Immunohistochemistry）分析肝細胞癌（hepatocellular carcinoma, HCC）的檢體，得到在肝細胞癌檢體中有71.4%的檢體呈現Pin1過度表現的結果；並且發現Pin1的表現量和臨床上的存活率也有相關性。Pin1的過度表現與肝細胞癌細胞株（HCC cell lines）的西方點墨法分析結果也是相符合的。我們也用反轉錄聚合連鎖反應（RT-PCR）檢測肝癌檢體的腫瘤部份（Tumor part）和成對的非腫瘤部份（Non-tumor part）之中Pin1 mRNA的表現量。我們得到的結果是，Pin1的過度表現可能導因於Pin1基因轉錄的上升（upregulation）。在大多數的肝細胞癌檢體當中，也被證明了Pin1之過度表現和β-catenin與Cyclin D1累積（accumulation）之間的關聯性；這個結果和之前與Pin1在乳癌癌化中可造成β-catenin及Cyclin D1累積的研究結果也有一致性。另外，我們也分析了肝炎病毒（hepatitis virus）慢性感染和Pin1過度表現之間的相關性。本論文的結果指出，Pin1過度表現可能導致肝細胞癌中β-catenin與Cyclin D1的累積，並且可能在肝細胞癌癌化過程中佔有重要的角色；因此Pin1的表現量可以做為肝細胞癌臨床診斷上的預後指標（prognostic marker），而且推測Pin1應是一個相當有潛力的肝細胞癌治療目標（therapeutic target）。

By Western blotting and immunohistochemical analyses, we have demonstrated that Pin1 was overexpressed in 71.4% of hepatocellular carcinoma (HCC) and its levels correlated with the clinical survival rate. This conclusion was supported by the results from examining Pin1 protein in HCC cancer cell lines. RT-PCR was performed to examine the Pin1 transcription level in tumor part and was compared with that in non-tumor part. Our results indicated that pin1 overexpression was due to the upregulation of Pin1 transcription. Interestingly, most of the cases with upregulation of Pin1 have been shown to correlate with β-catenin and Cyclin D1 accumulation in HCC specimens. These results were consistent with the previous studies that Pin1 caused β-catenin and Cyclin D1 elevation in breast cancer. The concordance between hepatitis virus chronic infection and Pin1 overexpression of HCC patients was also analysis. Taken together, these data indicated that Pin1 overexpression leading to β-catenin and Cyclin D1 accumulation might play a critical role in hepatocellular carcinogenesis and tumor progression. Pin1 levels therefore can be used as a prognostic marker for HCC, and our results suggested that Pin1 is a potential target for therapeutic intervention in hepatocellular carcinoma.

一、 前言………………………………………………………………1
二、 研究目標………………………………………………………....8
三、 材料與方法……………………………………………………....9
四、 結果……………………………………………………………..19
五、 討論……………………………………………………………..22
六、 參考文獻………………………………………………………..27
七、 附表……………………………………………………………..34
八、 附圖...…………………………………………………………...36
九、 附錄...…………………………………………………….……..49

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