2012-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/655995摘要：受器酪胺酸激脢是細胞生長的最重要訊息來源。癌症細胞常呈現同時活化多個受器酪胺酸激脢，因而影響癌細胞對分子標的療法的感受性。我們以受器酪胺酸激脢陣 列發現 Axl 是肝細胞癌細胞株最常見活化的受器酪胺酸激脢之一。我們將以下列實驗 研究 Axl 活化的機制，在癌症的功能，其下游分子機轉，及利用 Axl 抑制劑進行治療 肝細胞癌的可能。(1) 以反轉錄聚合脢鏈反應和免疫組織化學染色法研究肝細胞癌 Gas6 和 Axl 的 表現。(2) 把肝細胞癌細胞株的 Axl 過度表現或以 shRNA 敲毀 Axl 的表現，以 MTT 測 定，軟膠測定，和裸鼠注射試驗，證明 Axl 在肝細胞癌腫瘤生長的角色。 以 Boyden 試驗和傷口癒合試驗證明 Axl 在肝細胞癌腫瘤侵犯的角色。(3) 以 cDNA 微陣列等方法尋找 Gas6/Axl 途徑下游調控的基因。再以訊息傳遞途 徑的抑制分子，發光脢報導子試驗及染色質免疫沉澱法等實驗找出訊息傳 遞途徑。(4). 測試是否 Axl 的小分子抑制物可以加強 sorafenib 對肝細胞癌的抑制效果。<br> Abstract: Receptor tyrosine kinases (RTK) are the most important signal sources for cell growth. Cancer cells often activate several RTKs simultaneously, so that the response of cancer cell to molecularly targeted therapy is generally not very good. We have used RTK array to study the receptor tyrosine kinases activated in hepatocellular carcinoma (HCC) cell lines and found Axl is one of the RTKs most frequently activated in HCC. We will use the following experiments to study the mechanisms for activation of Axl, its functional roles in cancer, the downstream molecular mechanisms and the possibility of treating HCC with Axl inhibitors.(1). Use reverse transcriptase-polymerase chain reaction to study the expression of Gas6 and Axl in HCC.(2). Overexpression of Axl or knockdown the expression of Axl in HCC cell lines. We will use MTT assay, soft agar assay, and nude mice injection assay to prove the roles of Axl on the growth and tumorigenesis of HCC. We will also use Boyden chamber invasion assay and wound-healing assay to prove the roles of Axl in tumor invasion.(3). We will use cDNA array and other methods to identify the downstream targets of Axl/Gas6 pathway. Then we will use inhibitors of signal transduction pathway, luciferase reporter assay, and chromatin immunoprecipitation to identify the signal transduction mechanism.(4). We will test whether small inhibitors of Axl will enhance the inhibitory effect of sorafenib on HCC.