臺灣博碩士論文加值系統

合併台灣女性肺腺癌、轉移癌以及肺腺癌轉移至腦的三個基因微陣列分析資料，我們找到數個可能與癌轉移有關的基因。在本論文中，我針對一個目前研究尚在起步的基因KIAA0101進行研究，並驗證了在許多肺癌細胞株相對於正常肺纖維母細胞株都會有較高的KIAA0101蛋白質表現量。我們發現若抑制內生性KIAA0101蛋白質的表現，可明顯降低肺癌細胞株H1299的移行與侵入能力，而當過量表現KIAA0101時，則會增加肺腺癌細胞株CL1-0的移行與侵入的能力，說明了KIAA0101在癌細胞移行與侵入中可能扮演著部份的角色。而類似的結果也呈現在穩定表現KIAA0101的細胞株，相對於對照組，穩定過量表現KIAA0101會促進細胞的生長、增加細胞移行能力以及導致裸鼠的腫瘤生成。由於基端KIAA0101的截短蛋白質，幾乎喪失了促進細胞移行的能力，因此我們推測位於62~73胺基酸的PCNA binding motif或WW4 motif可能具有影響細胞移行的能力；此外，我更進一步發現藉由處理MMPs inhibitor (GM6001)會抑制KIAA0101所促進的細胞移行與侵入的能力以及穩定過量表現KIAA0101可降低E-cadherin蛋白質的表現量，因此，KIAA0101可能同時藉由調控EMT或是MMP的活性來影響細胞移行與侵入。綜合上述研究結果，本論文證明，KIAA0101能促進細胞移行、侵入以及腫瘤的生成，其將來極具潛力可做為一治療之標的或預後指標。
另外，我也證實KIAA0101在肝癌細胞亦具有在肺癌細胞相似的效果，即KIAA0101 mRNA 在肝癌組織呈現較高表現，且其蛋白質表現量亦與細胞移行呈現正相關。

By integrating three gene expression profile datasets, including pairwise female lung adenocarcinoma, secondary metastatic tumors vs. benign tumors, and lung adenocarinoma metastasis to brain, we have identified some novel metastasis-associated genes in lung adenocarcinoma. In this thesis, I focus on one poorly characterized gene, KIAA0101, and demonstrated the higher level of KIAA0101 expression is found in many lung cancer cell lines as compared to normal lung fibroblast. Knock-down the expression of KIAA0101 significantly decreased the migration and invasion ability of H1299 cells, while over-expression of KIAA0101 increased these events in CL1-0 cells, indicating KIAA0101 may play a role in promoting cell migration and invasion. Similar results are evidenced in stable KIAA0101 over-expression cells, which display higher growth rate and migration rate than vehicle cells, and promote tumor formation in nude mice in vivo. Since c-terminal truncate form of KIAA0101 (1~60 a.a.) almost lost it’s ability to promote cell migration, we speculate that PCNA binding or the WW4 motif, which is located between 62 ~73 a.a., may be crucial for KIAA0101 elicited cell migration. In addition, the presence of MMP inhibitor can decrease KIAA0101 induced cell migration and invasion and the decrease of E-cadherin expression was found in stable KIAA0101 over-expression cells, indicating KIAA0101 may concurrently regulate EMT event and MMP activity to affect cell migration and invasion. In accordance with the situation on lung cancer cell, the expression level of KIAA0101 is high in HCC tumor tissue and correlates with the migration abilities of HCC cell lines. Overall, this thesis demonstrated KIAA0101 could promote cell migration, invasion and tumorgenesis, which may be a promising target for cancer therapeutics or prognosis.

目 錄
中文摘要 ..………………………………………1
Abstract ..………………………………………2
縮寫表 ..………………………………………3
壹、序論 ..………………………………………4
貳、實驗材料 ………………………………………14
参、實驗方法 ………………………………………19
肆、結果 ………………………………………26
伍、討論 ………………………………………37
陸、圖表 ………………………………………44
柒、參考文獻 ………………………………………61

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