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研究生： 李佩蓉
Lee, Pei-Rong
論文名稱： 初級纖毛調控子宮內膜基質的分化
Primary cilium regulates uterine endometrial stroma differentiation
指導教授： 黃步敏
Huang, Bu-Miin
王家義
Wang, Chia-Yih
學位類別： 碩士
Master
系所名稱： 醫學院 - 細胞生物與解剖學研究所
Institute of Cell Biology and Anatomy
論文出版年： 2021
畢業學年度： 109
語文別： 英文
論文頁數： 45
中文關鍵詞： 蛻膜化 、初級纖毛 、子宮內膜基質 、cAMP 、孕激素受體
外文關鍵詞： decidualization, primary cilia, endometrial stroma, cAMP, progesterone receptor
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    • 懷孕流失是一種常見的產科併發症，約佔所有臨床孕婦的15％至25％。胚胎著床是懷孕的關鍵步驟，不好的著床會增加早期流產的風險。在妊娠早期，子宮內膜基質經歷了明顯的分子和形態變化，以進行胚胎著床和發育，這過程稱為子宮內膜蛻膜化。蛻膜化是由幾種細胞外因子引起的，像是前列腺素E2和鬆弛素會使細胞內cAMP的濃度增加。此時將會激活數個cAMP下游信號傳導，從而誘導蛻膜化基因的表達。另外，黃體酮激活黃體酮受體信號傳導與cAMP信號傳導具有協同作用，使蛻膜化作用最大化，從而可以正確著床。蛻膜化異常會導致著床失敗和流產，因此了解子宮內膜蛻膜化的機制對於增加胚胎著床率扮演著重要的角色。初級纖毛參與調控細胞的生長與分化，就像是細胞的天線會感應外在的環境，將接受到的訊息傳遞進入細胞，促使細胞做出回應。初級纖毛存在於許多人體細胞中，調控了許多細胞的分化，例如骨骼，脂肪，大腦，生殖器官，腎臟，視網膜和心臟。初級纖毛形成若缺陷會導致發育異常和疾病，統稱為纖毛缺損症候群。初級纖毛對於發育和分化很重要，但是初級纖毛在子宮內膜蛻膜化中所扮演的作用尚未被探討。在本篇研究中，我們證明了初級纖毛促進了子宮內膜蛻膜化。自細胞實驗中我們發現，當子宮基質細胞受到蛻膜化訊號刺激後，初級纖毛開始生長。利用藥理學抑制初級纖毛的生長或是利用遺傳基因耗竭破壞初級纖毛基因都會使蛻膜化作用減弱。然後，我們發現cAMP觸發了短纖毛形成。然而，孕激素受體信號傳導促進纖毛變長而成為功能性纖毛。最後我們也發現自小鼠子宮內膜中，初級纖毛的長度在交配過後隨著時間的增加而增長。因此，我們的研究揭示了初級纖毛在體內的新功能，並揭示了其潛在的分子機制。

    Pregnancy loss is a common obstetric complication and occurs in about 15-25% of all clinical pregnant women. Implantation of the embryo is a key step in pregnancy, and defective implantation increases the risk of early pregnancy loss. During early pregnancy, the uterine endometrial stroma undergoes dramatic molecular and morphological differentiation for embryo implantation and development, and this process is now known as endometrial decidualization. Decidualization is initiated with several extracellular factors such as prostaglandin E2 and relaxin followed by increasing intracellular concentration of cAMP. Several downstream cAMP signalings are activated thus inducing expressions of decidual markers genes. In addition, progesterone-activated progesterone receptor signaling cascade coordinately with cAMP signaling maximizes the decidualization. Defective decidualization leads to implantation failure and pregnancy loss. Thus, understanding the underlying mechanism by which decidual signaling promotes decidualization is important issue for promoting implantation rate. Primary cilia are involved in regulating the growth and differentiation; just like the antenna, primary cilia sense the external environment and transmit the received signals into the cell for maintaining physiological homeostasis. Primary cilia exist in many human cells and regulate the differentiation of many cells, such as bones, fat, brain, reproductive organs, kidneys, retina and heart; defects in primary cilium formation leads to several developmental abnormalities and diseases collectively termed ciliopathies. Primary cilium is important for development and differentiation, however, its role in endometrial decidualization has not been studied as yet before. In this study, we showed that decidualization was controlled by primary cilia. Upon decidual stimulation, primary cilia started to grow. Pharmacological inhibition of cilia formation or genetic depletion of ciliary genes attenuated decidualization. Then, we found cAMP triggered short cilia to grow; however, progesterone receptor signaling promoted short cilia became long cilia, also known as functional cilia. We also observed cilia became longer after copulation in mice model. Thus, our study uncovers the novel function of primary cilia in decidualization and deciphers the underlying molecular mechanism.

    口試合格證明書 I 中文摘要 II Abstract III Acknowledgement V Contents VI Abbreviations VIII Introduction 1 A.Endometrium: where the embryo implantation 1 B.Decidualization 2 C.Primary cilia 3 Material and method 7 Results 11 Discussion 16 Figures 19 Fig. 1. Decidualization in THESC cells. 19 Fig. 2. Decidual stimulation induces primary cilia formation in THESC cells. 20 Fig. 3. Decidual stimulation induces complete structure of primary cilia formation in THESC cells. 21 Fig. 4. Roscovitine inhibits decidualization and decidual-induced primary cilia in THESC cells. 22 Fig. 5. Depletion of primary cilia gene IFT88 inhibits primary cilia formation and decidualization. 23 Fig. 6. Depletion of primary cilia gene CEP164 inhibits primary cilia formation and decidualization. 24 Fig. 7. Co-treatment of cAMP and MPA induces primary cilia formation and decidualization in culture medium with normal serum. 26 Fig. 8. Treatment of cAMP induces short primary cilia in culture medium with charcoal -stripped serum. 28 Fig. 9. Two populations of cilia were observed during decidualization. 29 Fig. 10. Treatment of cAMP primes primary cilia formation in cells cultured with charcoal -stripped serum medium in THESC cells. 30 Fig. 11. Long cilia makes decidualization better. 31 Fig. 12. Co-treatment of cAMP and MPA induces decidualization in a dose dependent manner. 32 Fig. 13. Treatment of MPA did not induce PGR and phosphorylated PGR. 33 Fig. 14. Treatment of cAMP induces PGR but not phosphorylated PGR. 34 Fig. 15. Treatment of MPA/cAMP induces PGR and phosphorylated PGR. 35 Fig. 16. Treatment of cAMP induces expression of progesterone receptor, and co-treatment of cAMP and MPA induces expression of phosphorylated PGR. 36 Fig. 17. Depletion of progesterone receptor reduces long cilia formation. 37 Fig. 18. Depletion of progesterone receptor reduces long cilia formation. 38 Fig. 19. Endometrial stroma grows primary cilia during decidualization in vivo. 39 Fig. 20. A summary model shows that primary cilia regulates uterine endometrial stroma cells (ESCs) differentiation. 40 Reference 41

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