請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/36476
標題: | 週期性壓力刺激對類成骨母細胞基因表現的影響 Changes of gene expression in MG63 cell under cyclic compression force stimulation |
作者: | Chu-Yin Weng 翁竹音 |
指導教授: | 姚宗珍 |
關鍵字: | 基因微陣列,成骨母細胞,壓力,間質分解酵素-3, microarray,osteoblast,compression force,matrix metalloproteinase-3, |
出版年 : | 2005 |
學位: | 碩士 |
摘要: | 機械力量刺激對生長發育過程中骨骼重塑扮演重要的角色,但對力量刺激對細胞內所造成的基因變化影響,目前仍瞭解不多。為了篩選更多受壓力刺激影響之細胞基因,本研究利用類成骨母細胞株 (MG63 cell) 作為實驗對象,培養於3D膠原蛋白凝集體中,施予週期性壓力24小時。在帶有7680個基因的微陣列進行生物微陣列分析,經標準化後,共有43個基因在細胞受壓後有差異性表現,包括許多細胞凋亡、細胞週期調控、代謝、細胞增生、發炎反應等功能相關的基因,以及一些功能未知的基因。在進一步以半定量反轉錄-聚合酶鏈鎖反應 (RT-PCR) 分析,COX-2、ODC、MMP-3、MMP-10的表現上升,而ET-2、mIL-1RAcP的表現下降,符合微陣列之結果。以即時定量聚合酶連鎖反應 (Real-time PCR),發現COX-2 mRNA表現因壓力刺激提升2.27倍,而MMP-3 mRNA表現提升2.3倍。以西方點墨法分析細胞內MMP-3蛋白質的表現量,在壓力刺激下有微量上升的情形,但MMP-10蛋白質的表現量受機械壓力刺激後並無差異。以酵素活性分析法分析,MMP-3與MMP-10分解gelatin的活性微量受機械壓力刺激誘導增加表現。我們的結果支持著週期性壓力具有促進MG63細胞COX-2和MMP-3 mRNA表現,以及受壓後MMP-3蛋白質分泌也有增加。 It is widely accepted that mechanical loading is necessary to construct the architecture of bone and to maintain bone mass. Mechanical strain plays an important role in bone remodeling during growth and development. Several studies indicate that osteblastic cells respond to physical loading by transducing signals that alter gene expression patterns. However, the cellular gene expression that controls the tissue response to mechanical force remains unclear. To identify genes that may be affected by the compression force, we used a cyclic compression force applied to MG63 osteoblast-like cell in 3D collagen gel. We then performed a cDNA microarray analysis on an array contain 7680 gene-specific DNA fragments. After normalization, 43 genes had significant expression, including many cellular genes that involved in apoptosis, cell cycle regulator, cell proliferation, metabolism, inflammation and other cellular function were observed. Some selected genes were chosen for further study by RT-PCR analysis. Our results indicate that the expression of Cyclooxygenase-2 (COX-2), Ornithine Decarboxylase (ODC), Matrix metalloproteinase-3 and -10 (MMP-3 and MMP-10) was up regulated by compression force, whereas the expression of Endothelin-2 (ET-2), Membrane bound IL-1 receptor accessory protein (mIL-1RAcP) was down regulated, which were consistent with microarray data. We examined the protein expression of MMP-3 and MMP-10 by western blotting, and it showed that MMP-3 proteins were slightly elevated by compression force stimulation, whereas the expression of MMP-10 in compression group was similar to that of the control group. The protein expression of MMP-3 and MMP-10 was also detected by zymogram, and it showed that both proteins were slightly elevated by compression force stimulation. Our results showered that compression force induces COX-2 and MMP-3 mRNA expression, and MMP-3 protein level also increased. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/36476 |
全文授權: | 有償授權 |
顯示於系所單位: | 臨床牙醫學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-94-1.pdf 目前未授權公開取用 | 1.06 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。