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National Chung Hsing University Institutional Repository - NCHUIR > 生命科學院 > 生物醫學研究所 > 依資料類型分類 > 碩博士論文 >  14-3-3 zeta (YWHAZ)蛋白對於肺腺癌細胞 調控上皮塑性與轉移的重要性

Please use this identifier to cite or link to this item: http://nchuir.lib.nchu.edu.tw/handle/309270000/100946

標題: 14-3-3 zeta (YWHAZ)蛋白對於肺腺癌細胞 調控上皮塑性與轉移的重要性
The role of 14-3-3 zeta protein (YWHAZ) in regulating epithelial plasticity and lung adenocarcinoma cell metastasis
作者: 楊孟芳
Fang, Yang-Meng
Contributors: 楊泮池;俞松良
關鍵字: 14-3-3 zeta protein;lung adenocarcinoma
日期: 2009
Issue Date: 2012-09-04 10:49:35 (UTC+8)
Publisher: 生物醫學研究所
摘要: 肺癌,尤其是肺腺癌,是近年來具有高發生率和極高死亡率之惡性腫瘤,而癌轉移則是導致病患死亡的主因。癌轉移與細胞上皮塑性(epithelial plasticity)的改變有很大的關連。為找尋新的癌轉移相關基因,我們以競爭性基因體核酸雜交微陣列(CGHarray)分析不同侵襲與轉移能力肺癌細胞株之基因體中基因拷貝數的變異(CL1-0<CL1-5細胞株),並發現14-3-3zeta基因可能具有調控轉移的潛力。14-4-4zeta蛋白對於細胞功能的調控十分重要,包括訊息傳遞、細胞週期與凋亡等,然而在肺癌轉移所扮演的角色並不清楚。因此,本研究的目的主要為探討14-3-3zeta蛋白對肺腺癌細胞上皮塑性的調節與促進轉移之重要性。首先,確認14-3-3zeta蛋白在不同侵襲與移動能力之人類肺腺癌細胞株的表現量與細胞內分佈。接續,利用大量表現14-3-3zeta基因,探討14-3-3zeta蛋白對肺腺癌細胞型態的影響並進行與侵襲或移動相關功能之測試。為暸解14-3-3zeta蛋白是否調節肺腺癌細胞上皮塑性而影響細胞型態與功能,將評估14-3-3zeta蛋白對於上皮細胞轉型間葉細胞的形成(EMT)與G/F actin比例的分佈與改變。更進一步,鑑定14-3-3zeta蛋白改變上皮塑性過程中的交互作用分子與可能的調控機制。此外,以14-3-3zeta基因大量表現肺癌細胞株種植(seeding)於免疫不全鼠(SCID mice)說明14-3-3zeta蛋白在活體上對肺癌細胞移動之影響,證實14-3-3zeta是腫瘤發生與肺癌轉移所必須的分子物質。最後,檢測比較不同背景(不同時期)之肺腺癌病患檢體中,14-3-3zeta蛋白的表現量與分佈並加以病例資料分析,證實14-3-3zeta蛋白是否有潛力成為日後診斷肺腺癌的指標蛋白(marker protein),同時亦成為治療肺腺癌的標的蛋白(target protein)。由以上的結果,相信可以暸解14-3-3zeta蛋白對於人類肺腺癌轉移過程的影響,並希望對於癌症臨床治療有所貢獻。
Lung cancer, predominantly non-small-cell lung cancer (NSCLC), is the most common cause of cancer deaths worldwide, and metastasis is the major cause leading to mortality for cancer patients. The alteration of epithelial plasticity is crucial events during metastasis. To identify the novel metastasis-related genes, we used a method that simultaneously integrated microarray-based comparative genomic hybridization and affymetrix gene expression profiles to identifiy a potential candidate, 14-3-3zeta, in a lung cancer cell line model with different invasion capability (CL1-5 > CL1-0 cell line). 14-3-3zeta protein plays an important role in a wide variety of cellular processes, including signal transduction, cell cycle regulation and apoptosis. However, its role in lung cancer metastasis is unclear. This study will investigate the role of 14-3-3zeta protein in regulating epithelial plasticity and metastasis of lung adenocarcinoma cell. First, microarray data will be confirmed by real-time PCR and western blotting to demonstrate that the genomic copy number, mRNA and protein level of 14-3-3zeta in CL1-5 and CL1-0 lung caner cells. We also observe the distribution of 14-3-3zeta protein in subcellular localization. Second, we will overexpress or silence 14-3-3zeta in lung caner cells and estimate the ability of invasion, migration, as well as morphology. According to preliminary results, we presume that the protein may be involved in tumor metastasis via cytoskeleton rearrangement and regulating epithelial plasticity. To evaluate the alternation of epithelial plasticity, the amount of EMT marker,and G/F actin ratio will be performed by western blotting and immunofluorescence. In addition, we clarify the interaction molecule and modulating pathway of 14-3-3zeta protein. Furthermore, stable lung adenocarcinoma cell lines will be seeded onto immune-compromised (SCID) mice to suggest the influence of 14-3-3zeta protein. Consistent with an animal experiment, 14-3-3zeta is indeed essential for tumorigenesis and lung tumor metastasis. Finally, we check 14-3-3zeta expression in lung adenocarcinoma specimen with different stage, and analyze the survival and recurrent of 60 patients. These efforts may assist us to understand the role of 14-3-3zeta protein in human lung adenocarcinoma metastasis, and may offer some help in the treatment of lung cancer.
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