2013-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/705298摘要：先前在探討光動力致壓效應對腫瘤細胞轉移能力下降的研究中，我們發現CLIC4這一基因的改變，會影響MMP9的表現量與活性，並進而影響腫瘤細胞的轉移能力。雖然CLIC4一開始被發現為細胞內氯離子通道蛋白，但近年來在臨床腫瘤組織的研究中發現CLIC4與腫瘤的發展可能具有關聯性。然而，有關CLIC4與癌症發展與轉移之關係或分子調控機轉目前並不清楚。因此這一個計畫其目的在於探討CLIC4 在腫瘤組織發展及轉移中所扮演的角色及相關分子作用機轉。為此，我們將自兩個主軸來展開研究工作。第一個研究主軸上，我們將藉由細胞株及臨床病理組織檢體的研究，來分析CLIC4在腫瘤發展(tumorigenesis)及侵襲(invasiveness)中所扮演的可能角色。由於我們先前在細胞上的研究結果也顯示調控CLIC4之表現量將會直接對胞內MMP9造成影響，因此第二個研究主軸將在分子層次上探討CLIC4如何調控MMP9這一類影響癌轉移分子以及相關的控制機轉。為此我們擬定了下述幾個研究目標(specific aims)。第一、檢視CLIC4及MMP9在不具或有不同轉移能力細胞株中的表現量，並分析比較CLIC4在胞內位置是否和細胞轉移能力具關聯性；第二、利用shRNA或expressing plasmid來改變細胞內CLIC4的量，然後藉由 in vitro細胞及in vivo動物實驗來探討CLIC4對腫瘤發展、MMP9表現及癌細胞轉移侵襲能力的影響；第三、分析不同臨床腫瘤組織中，CLIC4及MMP9的表現量及其與腫瘤發展期別的關聯性；第四、找出CLIC4是藉由何種因子，來調控MMP9的表現；第五、探討CLIC4如何調控目標因子之作用機轉。 這一個計畫除了能清楚瞭解CLIC4在腫瘤組織發展及癌轉移分子的調控機制外，由於CLIC4表現量之變化與腫瘤的發展可能具有相當程度的關聯性，因此所得研究成果將來有潛力作為癌轉移生物標記或做為標靶治療的目標蛋白。<br> Abstract: Chloride intracellular channel 4 (CLIC4) belongs to the chloride intracellular channel family of proteins that may be involved with multiple cellular functions. However, recent studies in many human tumor types suggest that CLIC4 is an important participant in human cancer development. Previously, we have found that CLIC4 was significantly down regulated in PDT-derived variants with reduced ability of migration and invasion. Further studies showed that this reduced expression of CLIC4 could further down-regulate metalloproteinase-9 (MMP9) and result in the suppressed invasion of cancer cells. In this study, we intend to elucidate the role of CLIC4 in tumorigenesis and invasiveness. To fulfill this goal, we propose five specific aims for this three year research project. First, to examine the mRNA and protein contents of CLIC4 and MMP9 in normal and tumor cell lines with different invasive ability; Second, to elucidate the role of CLIC4 in tumor development in vitro and in vivo; Third, to investigate and correlate the expression level of CLIC4 and MMP9 in human neoplastic tissues with different stages of malignant progression; Forth, to investigate molecular profiling involved in CLIC4 mediated MMP9 expression; Fifth, to elucidate the molecular mechanisms involved in the CLIC4 mediated neoplastic development. These studies could clarify the unique role and molecular mechanism of CLIC4 during cancer pathogenesis. In the future, CLIC4 expression may be a useful addition to the diagnostic criteria and the growing list of novel targets for cancer therapy.CLIC4MMP-9腫瘤發展CLIC4MMP-9metastasis