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研究生:李脩琦
研究生(外文):Hsiu-ChiLee
論文名稱:雞卵蛋白上游驅動子轉錄因子II調控捲曲相關蛋白2在子宮內膜異位症發展過程之探討
論文名稱(外文):Characterization of the Functional Role of COUP-TFII Downstream Factor, Secreted Frizzle-Related Protein 2, in Endometriosis Development
指導教授:蔡少正
指導教授(外文):Shaw-Jenq 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:2012
畢業學年度:100
語文別:英文
論文頁數:43
中文關鍵詞:子宮內膜異位症雞卵蛋白上游驅動子轉錄因子II捲曲相關蛋白2抗細胞凋亡
外文關鍵詞:EndometriosisChicken ovalbumin upstream promoter transcription factor IISecreted frizzle related protein 2anti-apoptosis
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子宮內膜異位症是指子宮內膜組織在腹腔以外的地方生長,是十分常見並且病因複雜的婦科疾病,約有10~15%的生育婦女患有此疾病。臨床症狀常有:月經困難、性交疼痛、腹腔疼痛以及不孕等,這些因素明顯地減低婦女及其家人的生活品質。然而,子宮內膜異位症的病因目前還是不清楚,因此,此疾病的進程及其分子與細胞機轉是迫切地需要被研究並釐清。我們實驗室先前的研究指出,在異位的子宮內膜組織中,雞卵蛋白上游驅動子轉錄因子II的表現量相對於原位子宮內膜組織是較低的,這個現象意味著雞卵蛋白上游驅動子轉錄因子II可能參與在子宮內膜異位症的形成。究竟雞卵蛋白上游驅動子轉錄因子II在子宮內膜異位症的進程中有什麼功能,目前還是未知的。利用生物資訊以及微陣列的資料分析,我們認為捲曲相關蛋白2可能為雞卵蛋白上游驅動子轉錄因子II所調控的下游因子。過去認為捲曲相關蛋白2會與Wnt的受體競爭Wnt ,所以被視為是Wnt的拮抗物。研究結果顯示,在異位子宮內膜基質細胞中捲曲蛋白2 的傳訊者核醣核酸表現量明顯多於原位的子宮內膜基質細胞。藉由降低原位子宮內膜基質細胞的雞卵蛋白上游驅動子轉錄因子II的實驗,發現捲曲相關蛋白2 傳訊者核醣核酸表現量上升。給予正常子宮內膜基質細胞介白素-1β會抑制雞卵蛋白上游驅動子轉錄因子II的表現以及捲曲相關蛋白2的增加。此外,捲曲相關蛋白2能減少etoposide 所引起的Caspase-3 活化作用,避免細胞遭遇環境壓力所引起的細胞凋亡。綜合來說,在異位的子宮內膜基質細胞中雞卵蛋白上游驅動子轉錄因子II的表現量降低,使得捲曲相關蛋白2的增加而得以促進子宮內膜異位症的發展。
Endometriosis is a common gynecological disease in women. It is defined as the growth of endometria-like tissue outside the uterus. It typically occurs in 10-15% of reproductive age women. The symptoms of endometriosis are dysmenorrhea, dyspareunia, chronic pelvic pain, and infertility, which reduce life quality of affected women. However, the etiology is unclear thus it is necessary to investigate mechanisms involved in the development of endometriosis. Our previous study showed that the expression of chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) was reduced in endometriotic cells, suggesting a functional role of COUP-TFII in endometriosis. However, how COUP-TFII induces the pathological processes of endometriosis remains unknown. By using bioinformatics analysis and DNA microarray approaches, we identified secreted frizzle related protein 2 (SFRP2) as the candidate gene that may mediate loss-of-function of COUP-TFII-regulated endometrial pathogenesis. SFRP2 is considered as an antagonist for Wnt by competing the binding to Wnt receptor (frizzle). We demonstrated that SFRP2 was elevated in ectopic endometriotic stromal cells compared to eutopic endometrial stromal cells. Knockdown of COUP-TFII induced SFRP2 expression in eutopic stromal cells. Treatment of normal endometrial stromal cells with interleukin-1β inhibited COUP-TFII expression and resulted in SFRP2 upregulation. Furthermore, administration of SFRP2 attenuated etoposide-induced caspase-3 activation suggesting that SFRP2 can protect cells from stress-induced apoptosis. Taken together, loss of COUP-TFII in ectopic endometriotic stromal cells induces the expression of SFRP2, which may play an important role in the development of endometriosis.
Abstract in Chinese I
Abstract III
Acknowledgments V
Table of content VI
Introduction 1
Materials and methods 8
Reagents and buffers 8
Tissue collection 8
Cell culture 8
Isolation of total RNA 10
Reverse transcription polymerase chain reaction 11
Real-time PCR 11
Immunoblot 11
Bioinformatics tool 12
Statistical analysis 13
Results 14
1. COUP-TFII regulated candidate as stromal secretory protein 14
2. SFRP2 is up-regulated in ectopic stromal cell compare to eutopic stromal cell 15
3. Knockdown COUP-TFII up-regulates SFRP2 expression in mRNA level 15
4. SFRP2 may promote endometriotic cell survival 16
5. IL-1β represses COUP-TFII expression leading to induction of SFRP2 expression 16
Discussion 18
Figure 23
Fig. 1 Flowchart for the identification of COUP-TFII candidate genes 23
Fig. 2 Expression of SFRP2 is inversely correlates with levels of COUP-TFII 25
Fig. 3 Effect of COUP-TFII knockdown on SFRP2 RNA expression 26
Fig.4 Effect of SFRP2 on etoposide-induced apoptosis 27
Fig. 5 IL-1β inhibits COUP-TFII expression and induces SFRP2 expression 29
Appendix 30
Reagent and buffer used in cell culture 30
Reagent used in extraction of RNA or protein 31
Reagent and buffer used for western blot 32
Reference 36


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