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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 蔡丰喬(Feng-Chiao Tsai) | |
dc.contributor.author | Yin-Shan Chen | en |
dc.contributor.author | 陳吟珊 | zh_TW |
dc.date.accessioned | 2021-06-17T02:31:27Z | - |
dc.date.available | 2020-09-01 | |
dc.date.copyright | 2020-09-01 | |
dc.date.issued | 2020 | |
dc.date.submitted | 2020-08-17 | |
dc.identifier.citation | 1. Morgan, M. and S. McDonnell, Modulation of matrix metalloproteinases in trophoblast cell lines. Ann N Y Acad Sci, 1999. 878: p. 563-4. 2. Screen, M., et al., Cathepsin proteases have distinct roles in trophoblast function and vascular remodelling. Development, 2008. 135(19): p. 3311-20. 3. Silva, J.F. and R. Serakides, Intrauterine trophoblast migration: A comparative view of humans and rodents. Cell Adh Migr, 2016. 10(1-2): p. 88-110. 4. Goldman-Wohl, D. and S. Yagel, Regulation of trophoblast invasion: from normal implantation to pre-eclampsia. Mol Cell Endocrinol, 2002. 187(1-2): p. 233-8. 5. Tsai, F.C., et al., Ca2+ signaling in cytoskeletal reorganization, cell migration, and cancer metastasis. Biomed Res Int, 2015. 2015: p. 409245. 6. Wei, C., et al., Calcium flickers steer cell migration. Nature, 2009. 457(7231): p. 901-5. 7. Mo, P. and S. Yang, The store-operated calcium channels in cancer metastasis: from cell migration, invasion to metastatic colonization. Front Biosci (Landmark Ed), 2018. 23: p. 1241-1256. 8. Branch, K.M., D. Hoshino, and A.M. Weaver, Adhesion rings surround invadopodia and promote maturation. Biol Open, 2012. 1(8): p. 711-22. 9. Baczyk, D., J.C. Kingdom, and P. Uhlen, Calcium signaling in placenta. Cell Calcium, 2011. 49(5): p. 350-6. 10. Liu, Z., et al., Histamine enhances cytotrophoblast invasion by inducing intracellular calcium transients through the histamine type-1 receptor. Mol Reprod Dev, 2004. 68(3): p. 345-53. 11. Hache, S., et al., Alteration of calcium homeostasis in primary preeclamptic syncytiotrophoblasts: effect on calcium exchange in placenta. J Cell Mol Med, 2011. 15(3): p. 654-67. 12. Ghulmiyyah, L. and B. Sibai, Maternal mortality from preeclampsia/eclampsia. Semin Perinatol, 2012. 36(1): p. 56-9. 13. Vigil-De Gracia, P. and L. Ortega-Paz, Pre-eclampsia/eclampsia and hepatic rupture. Int J Gynaecol Obstet, 2012. 118(3): p. 186-9. 14. Ikechukwu, I.C., et al., Blood lead, calcium, and phosphorus in women with preeclampsia in Edo State, Nigeria. Arch Environ Occup Health, 2012. 67(3): p. 163-9. 15. Farzin, L. and F. Sajadi, Comparison of serum trace element levels in patients with or without pre-eclampsia. J Res Med Sci, 2012. 17(10): p. 938-41. 16. Hofmeyr, G.J., et al., Calcium supplementation during pregnancy for preventing hypertensive disorders and related problems. Cochrane Database Syst Rev, 2018. 10: p. CD001059. 17. Berridge, M.J., M.D. Bootman, and H.L. Roderick, Calcium signalling: dynamics, homeostasis and remodelling. Nat Rev Mol Cell Biol, 2003. 4(7): p. 517-29. 18. Berridge, M.J., P. Lipp, and M.D. Bootman, The versatility and universality of calcium signalling. Nat Rev Mol Cell Biol, 2000. 1(1): p. 11-21. 19. Collins, H.E., et al., STIM1/Orai1-mediated SOCE: current perspectives and potential roles in cardiac function and pathology. Am J Physiol Heart Circ Physiol, 2013. 305(4): p. H446-58. 20. Shen, W.W. and N. Demaurex, Morphological and functional aspects of STIM1-dependent assembly and disassembly of store-operated calcium entry complexes. Biochem Soc Trans, 2012. 40(1): p. 112-8. 21. Kiviluoto, S., et al., STIM1 as a key regulator for Ca2+ homeostasis in skeletal-muscle development and function. Skelet Muscle, 2011. 1(1): p. 16. 22. Gudlur, A., et al., Calcium sensing by the STIM1 ER-luminal domain. Nat Commun, 2018. 9(1): p. 4536. 23. Sharma, S., G. Godbole, and D. Modi, Decidual Control of Trophoblast Invasion. Am J Reprod Immunol, 2016. 75(3): p. 341-50. 24. Lee, C.L., et al., Role of Endoplasmic Reticulum Stress in Proinflammatory Cytokine-Mediated Inhibition of Trophoblast Invasion in Placenta-Related Complications of Pregnancy. Am J Pathol, 2019. 189(2): p. 467-478. 25. Wileman, T., et al., Depletion of cellular calcium accelerates protein degradation in the endoplasmic reticulum. J Biol Chem, 1991. 266(7): p. 4500-7. 26. Cui, Y., B. Zhu, and F. Zheng, Low-dose aspirin at </=16 weeks of gestation for preventing preeclampsia and its maternal and neonatal adverse outcomes: A systematic review and meta-analysis. Exp Ther Med, 2018. 15(5): p. 4361-4369. 27. Belcon, M.C., P.J. Rooney, and P. Tugwell, Aspirin and gastrointestinal haemorrhage: a methodologic assessment. J Chronic Dis, 1985. 38(1): p. 101-11. 28. Atallah, A., et al., Aspirin for Prevention of Preeclampsia. Drugs, 2017. 77(17): p. 1819-1831. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/68705 | - |
dc.description.abstract | 滋養層細胞在胎盤發育以及維持正常懷孕生理功能的過程中扮演著一個很重要的角色。當滋養層細胞的細胞遷移與侵襲作用不正常的時候,會造成一些妊娠失調的相關疾病如子癲前症、胎兒生長受限、植入性胎盤以及絨毛膜癌等等。然而,目前仍不清楚滋養層細胞是如何調控其細胞遷移與入侵的能力。先前的研究指出子癲前症患者血中的鈣離子濃度較正常孕婦低,且低血鈣症可能會使子癲前症的風險增加,表示鈣離子在這種疾病中相當關鍵。根據檢體資料庫的結果顯示在子癲前症患者的胎盤中STIM1的表現量較低,而STIM1是維持細胞內鈣離子恆定性的調節者。因此,我們推測STIM1可能對於滋養層細胞遷移與入侵有一定程度的影響,並進一步探究STIM1如何調控滋養層細胞遷移與侵襲。
由於滋養層細胞在進行細胞侵襲前需要先分泌一些proteases以降解細胞外基質,於是我們在滋養層細胞中knockdown STIM1,並利用protease protein array挑出兩個在knockdown STIM1後細胞外蛋白表現量明顯下降的兩個蛋白作為我們首要觀察的目標:MMP-2與cathepsin V。在3A-sub E與JAR中,細胞外MMP-2蛋白質表現量與活性在knockdown STIM1後都會降低。我們綜合先前文獻與實驗結果,推測這個現象可能與ER stress有關。我們假設在knockdown STIM1後ER鈣離子耗盡,進而產生ER stress與UPR,或者加速蛋白質的降解,使MMP-2蛋白質表現量與活性降低,最終導致細胞入侵的能力下降。為了驗證這個假說,我們以加藥方式調控ER內鈣離子與細胞質鈣離子的濃度。這些結果可能表明了ER內鈣離子對於調控protease的重要性遠大於細胞質鈣離子。一旦ER鈣離子耗盡,protease的細胞外蛋白質表現量與活性就會大幅降低。我們之後會進一步驗證knockdown STIM1後ER鈣離子耗盡對於滋養層細胞的影響,同時釐清STIM1在滋養層細胞中的功能,期許未來能藉由檢測STIM1基因表現量而提早對子癲前症進行預防與治療。 | zh_TW |
dc.description.abstract | Trophoblasts play an important role in placental development and maintaining normal physiological functions of pregnancy. Aberrant trophoblast migration and invasion cause gestational disorders including pre-eclampsia, fetal growth restriction, placenta accreta/increta and choriocarcinoma. However, it is still unclear how trophoblasts regulate their migration and invasion ability. Previous studies revealed that pre-eclampsia patients had lower blood Ca2+ level, and hypocalcemia can increase the risk of pre-eclampsia, indicating that calcium may play a key role in this disease. Placenta from patients with pre-eclampsia had reduced level of STIM1, which is the regulator of Ca2+ homeostasis. We therefore examined if STIM1 was involved in trophoblast migration and invasion.
Trophoblasts secrete some proteases to degrade extracellular matrix before they invade to the decidua or spiral arteries. Using protease protein array, we found that extracellular protein level of MMP-2 and cathepsin V decreased significantly by STIM1 knockdown in human trophoblast cell lines. We also verified that MMP-2 was reduced both in extracellular protein level and activity in 3A-sub E and JAR cells under STIM1 knockdown. Based on previous studies and our results, we speculate that this may be related to ER stress. Thus we hypothesize that STIM1 knockdown cause ER Ca2+ depletion, then induce ER stress and UPR or accelerate protein degradation, leading to decreased MMP-2 protein level and activity, finally resulting in the reduction of trophoblast invasion. To prove this hypothesis, we manipulated ER or cytosol Ca2+ pharmacologically. These data indicated that ER Ca2+ was more important than cytosol Ca2+ in protease regulation. Once ER Ca2+ depleted, extracellular protein level and activity of proteases reduced dramatically. We will further verify the above hypothesis, and also clarify the function of STIM1 in trophoblast, which may lead to the development of new therapeutic strategies in trophoblast-related diseases. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T02:31:27Z (GMT). No. of bitstreams: 1 U0001-1708202010001500.pdf: 4246073 bytes, checksum: 07b8664b58bf10da986b7ede63781476 (MD5) Previous issue date: 2020 | en |
dc.description.tableofcontents | 誌謝 2 中文摘要 3 Abstract 4 目錄 6 縮寫表 8 第一章、緒論 10 1-1 滋養層細胞在妊娠中的重要角色 10 1-1-1 滋養層細胞的主要功能 10 1-1-2 滋養層細胞相關疾病 11 1-2研究鈣離子以更加了解滋養層細胞調控細胞遷移與入侵的能力 12 1-2-1 鈣離子為影響細胞遷移與入侵之關鍵 12 1-2-2 鈣離子與子癲前症之間的關聯性 13 1-2-3 STIM1是維持細胞內鈣離子恆定性的調節者 13 1-2-4 STIM1水平與滋養層細胞的侵襲能力相關 15 研究動機 17 第二章、材料與方法 19 2-1 細胞培養 19 2-2 病毒製作與感染 19 2-3 Quantitative real time polymerase chain reaction (qPCR) 19 2-4 Western blot 20 2-5 Wound healing assay 20 2-6 Transwell assay 21 2-7 Protease protein array 22 2-8 Gelatin zymography assay 22 2-9 SOCE assay 22 第三章、結果 25 3-1 STIM1在滋養層細胞中的表現量與細胞間SOCE的差異 25 3-1-1 STIM1表現於滋養層細胞及絨毛膜癌細胞 25 3-1-2 細胞間SOCE的差異及利用knockdown STIM1方式降低SOCE 25 3-2 STIM1對於滋養層細胞的影響 27 3-2-1 STIM1 knockdown對於細胞遷移的影響 27 3-2-2 STIM1 knockdown對於protease表現量的影響 29 3-3 調控ER與細胞質鈣離子對於protease的影響 38 第四章、討論 42 4-1 驗證ER鈣離子耗盡對於protease功能的影響 42 4-2 STIM1調控滋養層細胞可能的機轉 44 4-3 STIM1調控滋養層細胞侵襲作用在臨床上的意義 45 4-4 細胞外蛋白質標準化方法 46 4-5 Transwell assay不穩定的原因 47 附錄 51 Matlab script 57 參考文獻 88 | |
dc.language.iso | zh-TW | |
dc.title | STIM1在滋養層細胞相關疾病中對於鈣離子調控蛋白質分解酵素表現與活性之探討 | zh_TW |
dc.title | STIM1 effects on trophoblast-related diseases: focusing on Ca2+-mediated protease expression and protease activity | en |
dc.type | Thesis | |
dc.date.schoolyear | 108-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 施景中(Jin-Chung Shih),龔秀妮(Hsiu-Ni Kung),陳文彬(Wen-Pin Chen),張美玲(Mei-Leng Cheong) | |
dc.subject.keyword | 滋養層細胞,子癲前症,細胞入侵, | zh_TW |
dc.subject.keyword | trophoblast,pre-eclampsia,STIM1,invasion,MMP-2, | en |
dc.relation.page | 89 | |
dc.identifier.doi | 10.6342/NTU202003687 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2020-08-18 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 藥理學研究所 | zh_TW |
顯示於系所單位: | 藥理學科所 |
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