跳到主要內容

臺灣博碩士論文加值系統

(18.97.14.86) 您好!臺灣時間:2025/02/08 01:07
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:陳妃琳
研究生(外文):Fei-Lin Chen
論文名稱:鈣離子及黏著分子的調控對癌細胞遷移之影響
論文名稱(外文):Interactions between Ca2+ and cell adhesion molecules during cancer cell migration
指導教授:蔡丰喬蔡丰喬引用關係
指導教授(外文):Feng-Chiao Tsai
口試日期:2017-07-20
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:藥理學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2017
畢業學年度:105
語文別:中文
論文頁數:89
中文關鍵詞:鈣離子store-operated Ca2+ entry (SOCE)粘著斑 (focal adhesionFA)細胞遷移癌症轉移
外文關鍵詞:Ca2+store-operated Ca2+ entry(SOCE)focal adhesioncell migrationcancer metastasis
相關次數:
  • 被引用被引用:0
  • 點閱點閱:189
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
細胞內鈣離子參與調控細胞增生、細胞內訊息傳遞以及細胞遷移,近幾年研究指出當細胞內鈣離子訊息傳遞失調時,會導致癌症的生成及轉移,而對於細胞內鈣離子的調控以及維持恆定性,其中store-operated Ca2+ entry (SOCE)扮演著重要調節腳色,然而對於SOCE是促進還是抑制癌細胞遷移仍然眾說紛紜,目前認為SOCE可能與粘著斑 (focal adhesion, FA)形成有關,就此影響細胞遷移能力。在先前研究發現讓癌細胞貼附於不同程度的細胞外基質上並抑制SOCE,在細胞移動速度上卻有著相反的結果,暗示著粘著斑與SOCE活性之間有著重要的平衡調控癌細胞遷移。
為了驗證上述想法我們藉由粘著斑與SOCE建立了細胞遷移速度的數學模型,並使用口腔鱗狀上皮癌細胞株SAS檢測模型的可靠性。具體做法為偵測SAS含有黏附因子及鈣離子調控蛋白的量,並應用遺傳學的方式使細胞過度表現或減少Paxillin (focal adhesion complex成員)及STIM1,或是用藥理學工具BTP2抑制SOCE情況下對SAS細胞遷移的影響,確立SAS在我們所建立的熱度圖上的位點。接著藉由同時過度表現不同程度的PXN及STIM1做細胞遷移實驗,追蹤每顆細胞個別帶有的PXN及STIM1量後畫出熱度圖,發現與我們所假設的模型相似,因此粘著斑與SOCE之間的平衡對癌細胞的遷移是重要的。
未來我們將(1)繼續驗證及修正此數學模型,並且能夠將此模型套用在其他癌細胞上,(2)透過高解析度活細胞影像平台,更了解癌細胞遷移之間SOCE及粘著斑平衡調控的機制,(3)將此模型進一步在動物及臨床上驗證,藉此預測癌症病人預後及抑制癌細胞轉移。
Ca2+ signaling regulates many cellular functions such as cell cycle progression and cell migration, which is also critical for cancer metastasis. Proper Ca2+ signaling requires store-operated Ca2+ entry (SOCE). However, there are controversies that SOCE may enhance or impede cancer cell migration from different literature reports. Recent studies suggested that SOCE-mediated Ca2+ signaling may modulate cell migration through interacting with focal adhesion (FA) complexes. Our previous study also showed that different cell-matrix adhesion strength and SOCE activities might differentially affect motility. These results indicate that FA and SOCE activities affect cancer cell migration in an integrated way.
To validate our speculations, we established a cell migration prediction heatmap using SOCE activities and focal adhesions. To verify our model, we suppressed or overexpressed Paxillin (a member of the focal adhesion complex) in SAS, an oral squamous cell carcinoma cell line. Also, we manipulated SOCE by altering STIM1 or treating the cells with BTP2, a SOCE inhibitor in SAS. Motilities of cells with above manipulations were measured to determine the original position of wild type SAS cell line on the heatmap. We then made each of the SAS cells expressing different levels of STIM1 and PXN, and measured their motilities in cell migration assays. These cells showed different migration activities as predicted in our prediction map. Therefore, our results support that proper balance between SOCE and focal adhesion is important for cancer cell migration.
In the future, we will (1) further validate and revise our model in other types of cancer cells, (2) use high resolution live-cell imaging platform to understand the mechanism how SOCE and FA regulate cancer cell migration, and (3) apply this model in animal experiments and clinical samples. With these approaches, this model will enable us to predict cancer prognosis precisely and to treat cancer metastasis efficiently.
目錄
誌謝 i
摘要 iii
Abstract iv
縮寫表 viii
第一章、 緒論 1
1-1 鈣離子影響細胞遷移 1
1-2 細胞遷移時鈣離子維持恆定 3
1-3 鈣離子影響癌症進程 5
1-4 SOCE影響癌細胞遷移透過黏著分子 7
1-5 研究動機與目的: 驗證細胞遷移熱度圖 9
第二章、 材料及方法 12
第三章、 結果 18
3-1 透過基因調控及藥物方式改變口腔癌細胞內鈣離子及黏著斑 18
3-1-1 細胞內Paxillin及STIM1表現量 18
3-1-2 利用lenti-virus系統操縱Paxillin及STIM1的表現 20
3-2 分別操縱鈣離子調控蛋白及黏著蛋白分子確立細胞在熱度圖上的位點 24
3-2-1 SAS分別overexpression及knockdown Paxillin降低細胞遷移速度 24
3-2-2 SAS分別overexpression及knockdown STIM1對細胞遷移速度的影響 27
3-3 驗證熱度圖的可行性透過改變SOCE及focal adhesion 30
3-3-1 同時在SAS增加STIM1及Paxillin表現量對細胞遷移之影響 30
第四章、 討論與結論 36
4-1 討論 36
4-1-1 細胞遷移熱度圖的完整性 36
4-1-2 在實際細胞遷移熱度圖中分成兩群細胞 38
4-1-3 機轉: 40
4-1-4 SAS overexpression STIM1表現量高組別的細胞knockdown PXN後STIM1表現量增加 42
4-1-5 臨床應用: 43
4-2 結論於未來展望 44
附錄 45
Matlab scripts 48
參考資料 86
Angelucci, A., and M. Bologna. 2007. Targeting Vascular Cell Migration as a Strategy for Blocking Angiogenesis: The Central Role of Focal Adhesion Protein Tyrosine Kinase Family. Current Pharmaceutical Design. 13:2129-2145.
Berridge, M.J. 1993. Inositol trisphosphate and calcium signalling. Nature. 361:315-325.
Chen, Y.-F., W.-T. Chiu, Y.-T. Chen, P.-Y. Lin, H.-J. Huang, C.-Y. Chou, H.-C. Chang, M.-J. Tang, and M.-R. Shen. 2011. Calcium store sensor stromal-interaction molecule 1-dependent signaling plays an important role in cervical cancer growth, migration, and angiogenesis. Proceedings of the National Academy of Sciences of the United States of America. 108:15225-15230.
Chigurupati, S., R. Venkataraman, D. Barrera, A. Naganathan, M. Madan, L. Paul, J.V. Pattisapu, G.A. Kyriazis, K. Sugaya, S. Bushnev, J.D. Lathia, J.N. Rich, and S.L. Chan. 2010. Receptor Channel TRPC6 Is a Key Mediator of Notch-Driven Glioblastoma Growth and Invasiveness. Cancer Research. 70:418-427.
Clapham, D.E.,2007. Calcium Signaling. Cell. 131:1047-1058.
Flourakis, M., V. Lehen''kyi, B. Beck, M. Raphaël, M. Vandenberghe, F. Vanden Abeele, M. Roudbaraki, G. Lepage, B. Mauroy, C. Romanin, Y. Shuba, R. Skryma, and N. Prevarskaya. 2010. Orai1 contributes to the establishment of an apoptosis-resistant phenotype in prostate cancer cells. Cell Death & Disease. 1:e75.
Franco, S.J., M.A. Rodgers, B.J. Perrin, J. Han, D.A. Bennin, D.R. Critchley, and A. Huttenlocher. 2004. Calpain-mediated proteolysis of talin regulates adhesion dynamics. Nat Cell Biol. 6:977-983.
Giannone, G., B.J. Dubin-Thaler, H.-G. Döbereiner, N. Kieffer, A.R. Bresnick, and M.P. Sheetz. 2004. Periodic Lamellipodial Contractions Correlate with Rearward Actin Waves. Cell. 116:431-443.
Grahovac, J., and A. Wells. 2014. Matrikine and matricellular regulators of EGF Receptor signaling on cancer cell migration and invasion. Laboratory investigation; a journal of technical methods and pathology. 94:31-40.
Levin, E.G. 2005. Cancer Therapy Through Control of Cell Migration. Current Cancer Drug Targets. 5:505-518.
Liou, J., M.L. Kim, W.D. Heo, J.T. Jones, J.W. Myers, J.E. Ferrell, and T. Meyer. 2005. STIM Is a Ca(2+) Sensor Essential for Ca(2+)-Store-Depletion-Triggered Ca(2+) Influx. Current biology : CB. 15:1235-1241.
Monteith, G.R., F.M. Davis, and S.J. Roberts-Thomson. 2012. Calcium channels and pumps in cancer: changes and consequences. J Biol Chem. 287:31666-31673.
Monteith, G.R., D. McAndrew, H.M. Faddy, and S.J. Roberts-Thomson. 2007. Calcium and cancer: targeting Ca2+ transport. Nat Rev Cancer. 7:519-530.
Ohga, K., R. Takezawa, Y. Arakida, Y. Shimizu, and J. Ishikawa. 2008. Characterization of YM-58483/BTP2, a novel store-operated Ca2+ entry blocker, on T cell-mediated immune responses in vivo. International Immunopharmacology. 8:1787-1792.
Peng, X., L.E. Cuff, C.D. Lawton, and K.A. DeMali. 2010. Vinculin regulates cell-surface E-cadherin expression by binding to β-catenin. Journal of Cell Science. 123:567-577.
Schmidt, S., G. Liu, G. Liu, W. Yang, S. Honisch, S. Pantelakos, C. Stournaras, A. Hönig, and F. Lang. 2014. Enhanced Orai1 and STIM1 expression as well as store operated Ca(2+) entry in therapy resistant ovary carcinoma cells. Oncotarget. 5:4799-4810.
Seccareccia, D. ,2010. Cancer-related hypercalcemia. Canadian Family Physician. 56:244-246.
Shaw, P.J., and S. Feske. . 2012. Physiological and pathophysiological functions of SOCE in the immune system. Frontiers in bioscience (Elite edition). 4:2253-2268.
Singh, A., M.E. Hildebrand, E. Garcia, and T.P. Snutch. 2010. The transient receptor potential channel antagonist SKF96365 is a potent blocker of low-voltage-activated T-type calcium channels. British Journal of Pharmacology. 160:1464-1475.
Stanisz, H., S. Saul, C.S.L. Müller, R. Kappl, B.A. Niemeyer, T. Vogt, M. Hoth, A. Roesch, and I. Bogeski. 2014. Inverse regulation of melanoma growth and migration by Orai1/STIM2-dependent calcium entry. Pigment Cell & Melanoma Research. 27:442-453.
Sun, J., F. Lu, H. He, J. Shen, J. Messina, R. Mathew, D. Wang, A.A. Sarnaik, W.C. Chang, M. Kim, H. Cheng, and S. Yang. 2014. STIM1- and Orai1-mediated Ca(2+) oscillation orchestrates invadopodium formation and melanoma invasion. J Cell Biol. 207:535-548.
Tkachenko, E., M. Sabouri-Ghomi, O. Pertz, C. Kim, E. Gutierrez, M. Machacek, A. Groisman, G. Danuser, and M.H. Ginsberg. 2011. Protein Kinase A Governs a RhoA-RhoGDI Protrusion-Retraction Pacemaker in Migrating Cells. Nature cell biology. 13:660-667.
Tsai, F.C., A. Seki, H.W. Yang, A. Hayer, S. Carrasco, S. Malmersjo, and T. Meyer. 2014. A polarized Ca2+, diacylglycerol and STIM1 signalling system regulates directed cell migration. Nat Cell Biol. 16:133-144.
Vashisht, A., M. Trebak, and R.K. Motiani. 2015. STIM and Orai proteins as novel targets for cancer therapy. A Review in the Theme: Cell and Molecular Processes in Cancer Metastasis. Am J Physiol Cell Physiol. 309:C457-469.
Wang, C., S. Chowdhury, M. Driscoll, C.A. Parent, S.K. Gupta, and W. Losert. 2014. The interplay of cell–cell and cell–substrate adhesion in collective cell migration. Journal of the Royal Society Interface. 11:20140684.
Wang, J.-Y., J. Sun, M.-Y. Huang, Y.-S. Wang, M.-F. Hou, Y. Sun, H. He, N. Krishna, S.-J. Chiu, S. Lin, S. Yang, and W.-C. Chang. 2015. STIM1 Overexpression Promotes Colorectal Cancer Progression, Cell Motility and COX-2 Expression. Oncogene. 34:4358-4367.
Yang, S., J.J. Zhang, and X.-Y. Huang. 2009a. Orai1 and STIM1 Are Critical for Breast Tumor Cell Migration and Metastasis. Cancer Cell. 15:124-134.
Yang, S., J.J. Zhang, and X.Y. Huang. 2009b. Orai1 and STIM1 are critical for breast tumor cell migration and metastasis. Cancer Cell. 15:124-134.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關期刊