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系統識別號 U0026-0812200915051696
論文名稱(中文) 轉化生長因子-β1對於子宮平滑肌細胞分化之調節作用
論文名稱(英文) The Regulation of Myometrial Cell Differentiation by Transforming Growth Factor-beta 1
校院名稱 成功大學
系所名稱(中) 生理學研究所
系所名稱(英) Department of Physiology
學年度 97
學期 1
出版年 98
研究生(中文) 李易儒
研究生(英文) Yiru Li
電子信箱 s3695101@mail.ncku.edu.tw
學號 s3695101
學位類別 碩士
語文別 中文
論文頁數 44頁
口試委員 指導教授-陳麗玉
口試委員-蔡少正
口試委員-江美治
中文關鍵字 轉化生長因子-β1  子宮肌層  分化作用  平滑肌細胞 
英文關鍵字 differentiation  Transforming growth factor-β1  smooth muscle cell  myometrium 
學科別分類
中文摘要 轉化生長因子-β1 (Transforming growth factor-β1; TGF-β1)是一具有許多功能的生長因子,可調控細胞的生長、分化以及胞外基質的分泌作用。TGF-β和其受器被偵測到在人類子宮肌層有表現情形,然而其扮演角色為何卻不得而知。過去有文獻指出TGF-β1可誘導間葉細胞表現出平滑肌細胞之分化指標蛋白SMα-actin (SMA) 和 SM myosin heavy chain (SM-MHC)。在此論文中,我們探討TGF-β1是否參與了子宮平滑肌細胞分化狀態的誘導。我利用人類子宮肌細胞進行實驗,子宮肌層檢體來源為醫院中做子宮全切除術的病患,我將子宮肌層小塊組織培養於含血清之培養液中以取得子宮平滑肌細胞。免疫螢光染色結果顯示,子宮肌層組織有表現 SMA和SM-MHC。在經過培養後,細胞失去了SM-MHC的表現,SMA的表現隨著細胞培養的代數而降低。細胞培養在無血清的狀態下,其SMA的表現增加。免疫細胞染色及西方墨點法結果均顯示,TGFβ對培養於無血清狀態下之細胞並未促進SMA之表現。2% FCS培養下,TGFβ降低細胞增生。經過TGF-β1處理2~4天後,可觀察到細胞呈現攤開狀;其SMA之表現量有增加情形。由以上結果顯示,TGF-β1可能參與了子宮平滑肌細胞的分化作用。
英文摘要 Transforming growth factor-β1 (TGF-β1) is an multifunctional cytokine that regulates cell growth, differentiation and matrix production. TGFβand its receptor were detected in human myometrium, however their roles are not known. It has been reported that TGFβ1 could induce the expression of smooth muscle cell (SMC) differentiation markers, such as SMα-actin (SMA) and SM myosin heavy chain (SM-MHC), in mesenchymal cells. In this study, we have investigated TGFβ1 might be involved in the induction of myometrial cell differentiation. Human myometrial tissues were obtained from the patients undergoing hysterectomy at the hospital. The tissue explants were cultured in 10% FCS for isolation of myometrial cells. Immunofluresence staining showed that myometrial tissues expressed SMA and SM-MHC. After culturing, SM-MHC expression in cells was lost. SMA expression in cells of advanced passage was reduced. Serum starvation arrested cell proliferation and increased SMA expression. Both immunofluresence staining and Western blot showed that TGFβ1 did not further increased SMA expression under serum free condition. We further compared cell proliferation and SMA in myometrial cells cultured in 2% FCS with the presence or absence of TGFβ1. The treatment with TGFβ1 for 2-4 days changed cell shape, decreased cell proliferation and increased SMA expression. All together, the results suggest that TGFβ1 may be involved in myometrial cell differentiation.
論文目次 目錄 I
圖表 III
Abstract 1
中文摘要 2
緒論 3
材料與方法 7
1.細胞培養 7
2.免疫組織化學染色法(Immunohistochemistry;IHC) 8
3.免疫螢光染色法(Immunofluresence staining) 9
(1) Smooth muscle α-actin (SMA) /Smooth muscle myosin
heavy chain (SM-MHC) 9
(2) 5’-Bromo-2’-deoxyuridine (BrdU) 10
4.西方墨點法(Western blot) 11
(1) 細胞處理及蛋白質萃取 11
(2) 蛋白質濃度測定 (Lowry assay) 12
(3) 蛋白質膠體電泳 (SDS-polyacrylamide gel
electrophoresis;SDS-PAGE) 13
(4) 蛋白質轉漬及免疫偵測 (Transfer and immunoblot) 13
藥品配製 15
結果 20
討論 23
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