機械循環拉伸對肺癌細胞功能的影響之研究;The effect of mechanical circulation stretching on the function of lung cancer cells

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Title:	機械循環拉伸對肺癌細胞功能的影響之研究;The effect of mechanical circulation stretching on the function of lung cancer cells
Authors:	黃元蓉;Huang, Yuan-Jung
Contributors:	系統生物與生物資訊研究所
Keywords:	肺癌細胞;機械循環拉伸;細胞侵襲;長片段非編碼RNA;lung cancer cells;mechanical circulation stretching;cell invasion;long non-coding RNA
Date:	2018-08-21
Issue Date:	2018-08-31 13:12:56 (UTC+8)
Publisher:	國立中央大學
Abstract:	在過去的文章我們知道機械循環拉伸會重新定位細胞骨架結構以及改變細胞的侵襲及爬行速率，所以本實驗的目的是探討機械循環拉伸對人類肺癌細胞功能的影響，而細胞功能的改變是由哪些基因在做調控。本實驗對象分別是人類肺腺癌細胞A549以及人類非小細胞肺癌細胞H322m，將這兩株細胞經由Boxer TM Cyclic Stretch Culture System (ATMS-Boxer)機械拉伸儀器進行拉伸實驗，拉伸的時間設定在0h、4h和24h，頻率設定在15%，0.5HZ。對拉伸與未拉伸的細胞分別做細胞侵襲、細胞遷移和細胞形成細胞群之能力以及DNA微陣列的分析。從實驗結果我們發現A549和H322m在經由機械循環拉伸四小時會促進細胞的侵襲能力(p-value < 0.05)，接著我們藉由DNA微陣列的分析，分析出CXCL1、 FOS、 CCL2、 CCL20、 BIRC3、TNFAIP3這6個基因在肺癌細胞拉伸四小時後與未拉伸的細胞相比有顯著性的差異(fold change > 2 or < -2 )。而這些基因大部分都是促進細胞侵襲能力，DNA微陣列的分析結果我們發現在拉伸四小時的細胞中都是表現量較高，這些拉力調控基因在路徑分析中發現可能藉由TNF信號通路來調控細胞的侵襲能力。;In the past, we known that mechanical cyclic stretching can induce reorganization of cytoskeleton and changed the cell migration and invasion. Therefor we identified the cyclic stretch-regulated gene expression in human lung cancer cell lines. Human lung cancer cell line A549 and H322m were subjected to cyclic strain (the frequency set in 15%, 0.5HZ) on 0h, 4h and 24h. We compared invasion assay, migration assay, colony formation and microarray assay in stretching and non-stretching groups. The results showed that the cell invasion in A549 and H322m were increased after 4hr stretching (p-value < 0.05). The microarray data showed that CXCL1, FOS, CCL2, CCL20, BIRC3, and TNFAIP3 were significant differences after 4hr stretching (fold change >2 or <-2). These genes have been reported to enhance cell invasion and these genes were cyclic stretch-regulated genes. Pathway enrichment analysis have found that these genes might regulate cells invasion through the TNF signaling pathway.
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