光甘草定抑制肺腺癌細胞移行、侵襲和血管新生之研究

The Inhibitory Effects of Glabridin on the Migration, Invasion, and Angiogenesis of Lung Adenocarcinoma Cells

蔡英明(Ying-Ming Tsai)

指導教授 ： 黃明賢

共同指導教授 ： 許雅玲(Ya-Ling Hsu)

高雄醫學大學/醫學院/醫學研究所/碩士(2009年)

https://doi.org/10.6832/KMU.2009.00063

摘要

迄今，肺癌仍為全世界重要癌症死亡原因之一，而非小細胞肺癌佔有約85%。過去癌症治療中，大部分的討論和研究重點都琢磨在癌細胞不正常增殖，而其他癌細胞之生理狀態，例如癌細胞不適當之移轉、侵襲等，並未獲得較多的重視。本論文重點是研究光甘草定在抑制肺癌細胞株A549移行、侵襲和血管新生的作用機轉。
本研究結果發現，光甘草定並未影響到肺癌細胞株Ａ549之增殖。但藉由傷口癒合的檢定和transwell移行檢定，證實光甘草定具有抑制A549移行能力；經由transwell侵襲檢定，證實光甘草定亦具有抑制A549侵襲能力，更進一步實驗結果證實，光甘草定並未能抑制MMP-2/9的分泌。而利用臍帶內皮細胞(human umbilical vein endothelial cells)之微血管管狀生成(capillary tube formation)實驗中，可以看到管狀構造形成的抑制現象。在動物實驗中，藉由血紅素檢定Matrigel plug中血紅素含量，可以證實光甘草定對於血管新生具有明顯抑制。
針對光甘草定抑制肺癌細胞移行、侵襲和血管新生的機轉，本研究發現在藥物處理下，αvβ3 integrin的表現以及FAK和Src的活化，呈現抑制的狀態。再者，本研究亦顯示光甘草定可有效抑制FAK/Src相互作用。而針對FAK/Src相關抑制移行、侵襲和血管新生的下游路徑，如Akt活化和myosin light chain (MLC)之磷酸化等，也出現相同的抑制現象。本實驗推測光甘草定抑制肺癌細胞移行、侵襲和血管新生之作用機轉是經由抑制αvβ3 integrin、FAK/Src以及其相關的路徑如Akt的活化與MLC磷酸化之訊息傳遞路徑。
結論，本篇研究證實，光甘草定對於肺癌細胞之移行、侵襲和血管新生都有抑制的效果，是故光甘草定為具有治療肺癌的潛能化合物。

關鍵字

肺癌 ； A549 ； 光甘草定 ； 移行 ； 侵襲 ； 轉移 ； 血管新生

並列摘要

Lung cancer is the most common cause of cancer-related death, with non-small-cell lung cancer (NSCLC) accounting for 80%–85% of all cases worldwide. In addition to uncontrolled cell proliferation and inappropriate cell survival, other defects in the acquisition of inappropriate migratory, invasive and angiogenic characteristics are less often studied. The aim of this study is to investigate the inhibitory mechanisms of glabridin on migration, invasion, and angiogenesis behaviors in lung cancer cell line A549.

The results reveal glabridin did not affect the cell proliferation. It confirmed that inhibitory migratory ability via wound-healing assay and transwell assay of migration, and inhibitory invasive ability via transwell assay of invasion despite no obvious effect on matrix metalloproteinases2/9 (MMP-2/9) expression. Angiogenesis was also suppressed from hemoglobin assay in vivo by hemoglobin level inside Matrigel plug and human umbilical vein endothelial cells (HUVECs) tube formation assay despite no obvious change in secreted vascular endothelial growth factor (VEGF) level.

The molecular mechanism was established which the αvβ3 integrin was inhibited by glabridin with down-regulation of focal adhesive kinase (FAK), and Src at specific phosphorylation sites. Other FAK/Src associated pathways such as Akt activation and myosin light chain (MLC) activation were also probed. Down-regulation of phosphorylated Akt and phosphorlated MLC were found. These experimental data favored the pathway was glabridin inhibited αvβ3 integrin, followed by down-regulation of FAK/Src, Akt, and MLC which led to inhibition of migration, invasion and angiogenesis.

In conclusion, this study implies glabridin could be of potential in treating lung cancer via inhibition of migration, invasion, and angiogenesis.

並列關鍵字

lung cancer ； A549 ； glabridin ； migration ； invasion ； metastasis ； angiogenesis

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