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研究生:陳秀梅
研究生(外文):Hsiu-Mei Chen
論文名稱:纖維母細胞生長因子-9在人類子宮內膜的表現及功能的調控機制
論文名稱(外文):The regulation of fibroblast growth factor-9 gene expression and physiological function in human uterine endometrium
指導教授:陳麗玉陳麗玉引用關係蔡少正
指導教授(外文):Lih-Yuh C. WingShaw-Jerq Tsai
學位類別:碩士
校院名稱:國立成功大學
系所名稱:生理學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:76
中文關鍵詞:纖維母細胞生長因子基質細胞上皮細胞
外文關鍵詞:epithelial cellsstromal cellsfibroblast growth factor
相關次數:
  • 被引用被引用:1
  • 點閱點閱:363
  • 評分評分:
  • 下載下載:68
  • 收藏至我的研究室書目清單書目收藏:0
纖維母細胞生長因子-9 ( fibroblast growth factor-9)是一種能刺激數種神經細胞和前列腺細胞增生及存活的蛋白質生長因子。然而,纖維母細胞生長因子-9在雌性生殖道方面是否有類似的功能則尚未被研究。我的論文主要是要探討纖維母細胞生長因子-9在人類子宮內膜所扮演的角色。我的研究結果指出,纖維母細胞生長因子-9在正常的子宮內膜大量的表現,其表現在增生期的後期達到最高峰,而這個時期正是動情激素大量分泌及子宮內膜快速增生的時期,這些結果顯示三者之間有密切的關連性。使用反轉錄脢聚合脢鏈結反應的實驗結果發現,纖維母細胞生長因子-9主要表現在子宮內膜的基質細胞。此外我也發現纖維母細胞生長因子-9的受體,即纖維母細胞生長因子受體-2及-3,可表現在上皮細胞及基質細胞中。 纖維母細胞生長因子受體-2IIIc及-3IIIc在子宮內膜基質細胞的表現最多,纖維母細胞生長因子受體-2IIIb主要表現在上皮細胞,而纖維母細胞生長因子受體-3IIIb則是在基質細胞及上皮細胞都有表現。纖維母細胞生長因子-9刺激基質細胞增生的能力隨著濃度上升而提高,相反的,它卻無法刺激上皮細胞的增生。纖維母細胞生長因子-9可以刺激基質細胞內cyclin A及cyclin D1的產生。我利用不同的磷酸化脢抑制劑來探討纖維母細胞生長因子-9作用之訊息傳遞途徑,實驗結果顯示纖維母細胞生長因子-9刺激基質細胞增生是透過PI3 kinase以及MAPK這兩條途徑。此外,以動情激素處理可以刺激基質細胞表現纖維母細胞生長因子-9以及刺激基質細胞的增生,而此動情激素刺激基質細胞增生的功能可以被纖維母細胞生長因子-9的抗體所抑制。總結而言,我的研究證實,纖維母細胞生長因子-9是一種受到動情激素所調控的基質細胞自泌生長因子,而且在子宮內膜增生時期扮演著相當重要的角色。
Fibroblast growth factor-9 (FGF-9) is a potent mitogen and survival factor for numerous nerve cells and prostatic cells. However, the significance of FGF-9 in female reproductive tract has not been investigated. The objective of this study is to examine the role of FGF-9 in human uterine endometrium. My results show that FGF-9 is expressed at high level in normal uterine endometrium, especially during the late proliferative phase, which is coincident with the rise of estradiol and the time of uterine endometrium proliferation. Using RT-PCR analysis, I found that FGF-9 was produced primarily by endometrial stromal cells. High affinity receptors of FGF-9 were detected in both epithelial and stromal cells with distinct patterns. FGFR2IIIc and FGFR3IIIc were the most abundant isoforms in endometrial stromal cells. FGFR2IIIb was markedly expressed in endometrial epithelial cells whereas FGFR3IIIb was found in both epithelial and stromal cells. Treatment with FGF-9 induced endometrial stromal proliferation in a dose-dependent manner but had no substantial effect on epithelial cells. In accordance with this this result, cyclin D1 and cyclin A were induced by thetreatment with FGF-9 in stromal cells. Using selective protein kinase inhibitors demonstrated that FGF-9 induced cell proliferation was mediated via phosphatidylinositol 3-kinase and mitogen-activated protein kinase signaling pathways. Treatment of stromal cells with 17��-estrodiol stimulated FGF-9 expression and cell proliferation. Furthermore, the mitogenic effect of estrogen was inhibited by co-treatment with anti-FGF-9 antibody. In conclusion, the results of my study have demonstrated, for the first time, that FGF-9 is an autocrine estromedin endometrial stromal growth factor that plays significant roles in cyclic proliferation of uterine endometrium.
目錄
致謝….1
目錄….2
圖表….3
英文摘要….4
中文摘要….5
緒論….6
實驗方法…19
a. 正常子宮內膜組織上皮細胞以及基質細胞的分離…19
b. 培養之子宮內膜基質細胞及上皮細胞純度鑑定方法…20
c. 細胞數的計算及細胞存活率之鑑定….21
d. RNA之純化與分離…21
e. 反轉錄聚合脢連鎖反應(RT-PCR)…22
f. 以免疫染色法檢測纖維母細胞成長因子受體(FGFR)…23
g. 細胞攝取3H-thymidine後保留放射性同位素之能力…24
h. 蛋白質濃度分析(Lowry assay)…25
i. SDS-PAGE 蛋白質電泳…25
藥品配製…28
結果…36
討論…43
參考文獻…51
圖…57
作者簡歷…76
參考文獻

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