酪氨酸激酶調控跟細胞生長、移動性、及分化相關的訊息傳遞路徑。在正常細胞中，酪氨酸激酶的活性受到嚴密的監控；然而癌細胞會誘發酪氨酸激酶過度的活化以促進癌化現象。目前有部分的酪氨酸激酶抑制劑使用於癌症病患的臨床治療。我們實驗中所使用的新型酪氨酸激酶抑制劑，1J-309，具有抑制多種酪氨酸激酶的功效，對於血管內皮生長因子受器(VEGFRs) 也有其抑制功效。在我們的實驗中發現1J-309有效的抑制VEGFR3+/VEGF-C+的A549人類肺癌細胞的細胞生長能力並經由降低細胞週期蛋白依賴型激酶CDK1及週期蛋白cyclin B1的表現導致細胞生長週期停止在G2/M phase。在長時間的藥物培養下，1J-309造成A549細胞大量死亡。雖然1J-309可早期抑制CDK1蛋白質表現，卻不會影響CDK1其核醣核酸(RNA)的表現及CDK1的蛋白質穩定性。另一方面，1J-309可以大幅降低A549肺癌細胞的移動性以及減緩A549細胞對gelatin的分解進而抑制其侵犯性。我們也發現1J-309大幅抑制AKT及p38 MAPK的訊息傳遞路徑並且在低濃度時造成腫瘤抑制基因p53的蛋白質高度表現。本實驗證實1J-309可有效的抑制癌細胞的細胞生長經由引發生長週期停止在G2/M phase、降低其移動性及侵犯性以及大量誘發腫瘤抑制基因p53的表現。

Tyrosine kinases regulate fundamental signal pathways in cells including cell proliferation, motility, and differentiation. The kinase activity is tightly controlled in normal cells but is usually excessive activated in cancers. Several tyrosine kinase inhibitors are used in cancer therapies nowadays. Our novel tyrosine kinase inhibitor, 1J-309, is a multiple kinase inhibitor that targets several receptors including vascular endothelial growth factor receptors (VEGFRs). We find 1J-309 dramatically reduces cell proliferation of VEGFR3+/VEGF-C+ A549 human lung cancer cells by decreasing the expression of CDK1 and cyclin B1 following growth arrest at G2/M phase. After long term drug treatment, 1J-309 causes cell death. Moreover, 1J-309 represses CDK1 expression at early stage but it does not change CDK1 RNA expression and protein stability. Additionally, 1J-309 significantly decreases the migration ability of A549 cells. 1J-309 also reduces gelatin-related invasion potency. The AKT and p38 MAPK activity are significantly repressed by 1J-309 and it dramatically drives the expression of tumor suppressor, p53, at low-dose treatment. Our results demonstrate that 1J-309 significantly attenuates cell proliferation by inducing G2/M growth arrest, reduces the invasion and migration potency, and promotes a dramatic increase of p53 in A549 cells.

Abstract (in Chinese) -----------------------------------------------------------------------------ii
Abstract (in English) -----------------------------------------------------------------------------iii
Introduction -----------------------------------------------------------------------------------------1
Specific aim ----------------------------------------------------------------------------------------7
Material and methods ---------------------------------------------------------------------------8
Results ---------------------------------------------------------------------------------------------19
Discussion -----------------------------------------------------------------------------------------26
Figures and table ---------------------------------------------------------------------------------31
Supplementary information --------------------------------------------------------------------49
References -----------------------------------------------------------------------------------------58

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