請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30346
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 李心予(Hsinyu Lee) | |
dc.contributor.author | Yu-Ting Huang | en |
dc.contributor.author | 黃鈺婷 | zh_TW |
dc.date.accessioned | 2021-06-13T02:01:42Z | - |
dc.date.available | 2007-07-16 | |
dc.date.copyright | 2007-07-16 | |
dc.date.issued | 2007 | |
dc.date.submitted | 2007-07-06 | |
dc.identifier.citation | Albelda, S.M., Muller, W.A., Buck, C.A. and Newman, P.J. (1991) Molecular and cellular properties of PECAM-1 (endoCAM/CD31): a novel vascular cell-cell adhesion molecule, The Journal of cell biology, 114, 1059-1068.
An, S., Goetzl, E.J. and Lee, H. (1998) Signaling mechanisms and molecular characteristics of G protein-coupled receptors for lysophosphatidic acid and sphingosine 1-phosphate, J Cell Biochem Suppl, 30-31, 147-157. Anliker, B. and Chun, J. (2004) Cell surface receptors in lysophospholipid signaling, Seminars in cell & developmental biology, 15, 457-465. Anliker, B. and Chun, J. (2004) Lysophospholipid G protein-coupled receptors, The Journal of biological chemistry, 279, 20555-20558. Ayalon, O., Sabanai, H., Lampugnani, M.G., Dejana, E. and Geiger, B. (1994) Spatial and temporal relationships between cadherins and PECAM-1 in cell-cell junctions of human endothelial cells, The Journal of cell biology, 126, 247-258. Bird, I.N., Taylor, V., Newton, J.P., Spragg, J.H., Simmons, D.L., Salmon, M. and Buckley, C.D. (1999) Homophilic PECAM-1(CD31) interactions prevent endothelial cell apoptosis but do not support cell spreading or migration, Journal of cell science, 112 ( Pt 12), 1989-1997. Biswas, P., Canosa, S., Schoenfeld, J., Schoenfeld, D., Tucker, A. and Madri, J.A. (2003) PECAM-1 promotes beta-catenin accumulation and stimulates endothelial cell proliferation, Biochemical and biophysical research communications, 303, 212-218. Bogen, S., Pak, J., Garifallou, M., Deng, X. and Muller, W.A. (1994) Monoclonal antibody to murine PECAM-1 (CD31) blocks acute inflammation in vivo, The Journal of experimental medicine, 179, 1059-1064. Cao, G., O'Brien, C.D., Zhou, Z., Sanders, S.M., Greenbaum, J.N., Makrigiannakis, A. and DeLisser, H.M. (2002) Involvement of human PECAM-1 in angiogenesis and in vitro endothelial cell migration, American journal of physiology, 282, C1181-1190. Cao, M.Y., Huber, M., Beauchemin, N., Famiglietti, J., Albelda, S.M. and Veillette, A. (1998) Regulation of mouse PECAM-1 tyrosine phosphorylation by the Src and Csk families of protein-tyrosine kinases, The Journal of biological chemistry, 273, 15765-15772. Cepinskas, G., Savickiene, J., Ionescu, C.V. and Kvietys, P.R. (2003) PMN transendothelial migration decreases nuclear NFkappaB in IL-1beta-activated endothelial cells: role of PECAM-1, The Journal of cell biology, 161, 641-651. Cicmil, M., Thomas, J.M., Sage, T., Barry, F.A., Leduc, M., Bon, C. and Gibbins, J.M. (2000) Collagen, convulxin, and thrombin stimulate aggregation-independent tyrosine phosphorylation of CD31 in platelets. Evidence for the involvement of Src family kinases, The Journal of biological chemistry, 275, 27339-27347. DeLisser, H.M., Chilkotowsky, J., Yan, H.C., Daise, M.L., Buck, C.A. and Albelda, S.M. (1994) Deletions in the cytoplasmic domain of platelet-endothelial cell adhesion molecule-1 (PECAM-1, CD31) result in changes in ligand binding properties, The Journal of cell biology, 124, 195-203. DeLisser, H.M., Christofidou-Solomidou, M., Strieter, R.M., Burdick, M.D., Robinson, C.S., Wexler, R.S., Kerr, J.S., Garlanda, C., Merwin, J.R., Madri, J.A. and Albelda, S.M. (1997) Involvement of endothelial PECAM-1/CD31 in angiogenesis, The American journal of pathology, 151, 671-677. Gao, C., Sun, W., Christofidou-Solomidou, M., Sawada, M., Newman, D.K., Bergom, C., Albelda, S.M., Matsuyama, S. and Newman, P.J. (2003) PECAM-1 functions as a specific and potent inhibitor of mitochondrial-dependent apoptosis, Blood, 102, 169-179. Garcia, J.G., Liu, F., Verin, A.D., Birukova, A., Dechert, M.A., Gerthoffer, W.T., Bamberg, J.R. and English, D. (2001) Sphingosine 1-phosphate promotes endothelial cell barrier integrity by Edg-dependent cytoskeletal rearrangement, The Journal of clinical investigation, 108, 689-701. Graler, M.H., Bernhardt, G. and Lipp, M. (1998) EDG6, a novel G-protein-coupled receptor related to receptors for bioactive lysophospholipids, is specifically expressed in lymphoid tissue, Genomics, 53, 164-169. Gratzinger, D., Barreuther, M. and Madri, J.A. (2003) Platelet-endothelial cell adhesion molecule-1 modulates endothelial migration through its immunoreceptor tyrosine-based inhibitory motif, Biochemical and biophysical research communications, 301, 243-249. Gratzinger, D., Canosa, S., Engelhardt, B. and Madri, J.A. (2003) Platelet endothelial cell adhesion molecule-1 modulates endothelial cell motility through the small G-protein Rho, Faseb J, 17, 1458-1469. Hla, T. and Maciag, T. (1990) An abundant transcript induced in differentiating human endothelial cells encodes a polypeptide with structural similarities to G-protein-coupled receptors, The Journal of biological chemistry, 265, 9308-9313. Ilan, N., Cheung, L., Miller, S., Mohsenin, A., Tucker, A. and Madri, J.A. (2001) Pecam-1 is a modulator of stat family member phosphorylation and localization: lessons from a transgenic mouse, Developmental biology, 232, 219-232. Ilan, N., Cheung, L., Pinter, E. and Madri, J.A. (2000) Platelet-endothelial cell adhesion molecule-1 (CD31), a scaffolding molecule for selected catenin family members whose binding is mediated by different tyrosine and serine/threonine phosphorylation, The Journal of biological chemistry, 275, 21435-21443. Ilan, N., Mahooti, S., Rimm, D.L. and Madri, J.A. (1999) PECAM-1 (CD31) functions as a reservoir for and a modulator of tyrosine-phosphorylated beta-catenin, Journal of cell science, 112 Pt 18, 3005-3014. Im, D.S., Heise, C.E., Ancellin, N., O'Dowd, B.F., Shei, G.J., Heavens, R.P., Rigby, M.R., Hla, T., Mandala, S., McAllister, G., George, S.R. and Lynch, K.R. (2000) Characterization of a novel sphingosine 1-phosphate receptor, Edg-8, The Journal of biological chemistry, 275, 14281-14286. Jackson, D.E., Kupcho, K.R. and Newman, P.J. (1997) Characterization of phosphotyrosine binding motifs in the cytoplasmic domain of platelet/endothelial cell adhesion molecule-1 (PECAM-1) that are required for the cellular association and activation of the protein-tyrosine phosphatase, SHP-2, The Journal of biological chemistry, 272, 24868-24875. Jackson, D.E., Ward, C.M., Wang, R. and Newman, P.J. (1997) The protein-tyrosine phosphatase SHP-2 binds platelet/endothelial cell adhesion molecule-1 (PECAM-1) and forms a distinct signaling complex during platelet aggregation. Evidence for a mechanistic link between PECAM-1- and integrin-mediated cellular signaling, The Journal of biological chemistry, 272, 6986-6993. Kogata, N., Masuda, M., Kamioka, Y., Yamagishi, A., Endo, A., Okada, M. and Mochizuki, N. (2003) Identification of Fer tyrosine kinase localized on microtubules as a platelet endothelial cell adhesion molecule-1 phosphorylating kinase in vascular endothelial cells, Molecular biology of the cell, 14, 3553-3564. Le Stunff, H., Milstien, S. and Spiegel, S. (2004) Generation and metabolism of bioactive sphingosine-1-phosphate, Journal of cellular biochemistry, 92, 882-899. Lee, H., Goetzl, E.J. and An, S. (2000) Lysophosphatidic acid and sphingosine 1-phosphate stimulate endothelial cell wound healing, American journal of physiology, 278, C612-618. Lee, H., Lin, C.I., Liao, J.J., Lee, Y.W., Yang, H.Y., Lee, C.Y., Hsu, H.Y. and Wu, H.L. (2004) Lysophospholipids increase ICAM-1 expression in HUVEC through a Gi- and NF-kappaB-dependent mechanism, American journal of physiology, 287, C1657-1666. Lee, M.J., Thangada, S., Claffey, K.P., Ancellin, N., Liu, C.H., Kluk, M., Volpi, M., Sha'afi, R.I. and Hla, T. (1999) Vascular endothelial cell adherens junction assembly and morphogenesis induced by sphingosine-1-phosphate, Cell, 99, 301-312. Lee, O.H., Kim, Y.M., Lee, Y.M., Moon, E.J., Lee, D.J., Kim, J.H., Kim, K.W. and Kwon, Y.G. (1999) Sphingosine 1-phosphate induces angiogenesis: its angiogenic action and signaling mechanism in human umbilical vein endothelial cells, Biochemical and biophysical research communications, 264, 743-750. Liao, F., Huynh, H.K., Eiroa, A., Greene, T., Polizzi, E. and Muller, W.A. (1995) Migration of monocytes across endothelium and passage through extracellular matrix involve separate molecular domains of PECAM-1, The Journal of experimental medicine, 182, 1337-1343. Limaye, V., Li, X., Hahn, C., Xia, P., Berndt, M.C., Vadas, M.A. and Gamble, J.R. (2005) Sphingosine kinase-1 enhances endothelial cell survival through a PECAM-1-dependent activation of PI-3K/Akt and regulation of Bcl-2 family members, Blood, 105, 3169-3177. Lin, C.I., Chen, C.N., Chen, J.H. and Lee, H. (2006) Lysophospholipids increase IL-8 and MCP-1 expressions in human umbilical cord vein endothelial cells through an IL-1-dependent mechanism, Journal of cellular biochemistry, 99, 1216-1232. Lu, T.T., Barreuther, M., Davis, S. and Madri, J.A. (1997) Platelet endothelial cell adhesion molecule-1 is phosphorylatable by c-Src, binds Src-Src homology 2 domain, and exhibits immunoreceptor tyrosine-based activation motif-like properties, The Journal of biological chemistry, 272, 14442-14446. Lu, T.T., Yan, L.G. and Madri, J.A. (1996) Integrin engagement mediates tyrosine dephosphorylation on platelet-endothelial cell adhesion molecule 1, Proceedings of the National Academy of Sciences of the United States of America, 93, 11808-11813. MacLennan, A.J., Browe, C.S., Gaskin, A.A., Lado, D.C. and Shaw, G. (1994) Cloning and characterization of a putative G-protein coupled receptor potentially involved in development, Molecular and cellular neurosciences, 5, 201-209. Mamdouh, Z., Chen, X., Pierini, L.M., Maxfield, F.R. and Muller, W.A. (2003) Targeted recycling of PECAM from endothelial surface-connected compartments during diapedesis, Nature, 421, 748-753. Masuda, M., Osawa, M., Shigematsu, H., Harada, N. and Fujiwara, K. (1997) Platelet endothelial cell adhesion molecule-1 is a major SH-PTP2 binding protein in vascular endothelial cells, FEBS letters, 408, 331-336. Muller, A.M., Hermanns, M.I., Skrzynski, C., Nesslinger, M., Muller, K.M. and Kirkpatrick, C.J. (2002) Expression of the endothelial markers PECAM-1, vWf, and CD34 in vivo and in vitro, Experimental and molecular pathology, 72, 221-229. Muller, W.A., Weigl, S.A., Deng, X. and Phillips, D.M. (1993) PECAM-1 is required for transendothelial migration of leukocytes, The Journal of experimental medicine, 178, 449-460. Nakada, M.T., Amin, K., Christofidou-Solomidou, M., O'Brien, C.D., Sun, J., Gurubhagavatula, I., Heavner, G.A., Taylor, A.H., Paddock, C., Sun, Q.H., Zehnder, J.L., Newman, P.J., Albelda, S.M. and DeLisser, H.M. (2000) Antibodies against the first Ig-like domain of human platelet endothelial cell adhesion molecule-1 (PECAM-1) that inhibit PECAM-1-dependent homophilic adhesion block in vivo neutrophil recruitment, J Immunol, 164, 452-462. Newman, D.K., Hamilton, C. and Newman, P.J. (2001) Inhibition of antigen-receptor signaling by Platelet Endothelial Cell Adhesion Molecule-1 (CD31) requires functional ITIMs, SHP-2, and p56(lck), Blood, 97, 2351-2357. Newman, P.J., Berndt, M.C., Gorski, J., White, G.C., 2nd, Lyman, S., Paddock, C. and Muller, W.A. (1990) PECAM-1 (CD31) cloning and relation to adhesion molecules of the immunoglobulin gene superfamily, Science (New York, N.Y, 247, 1219-1222. Newman, P.J., Hillery, C.A., Albrecht, R., Parise, L.V., Berndt, M.C., Mazurov, A.V., Dunlop, L.C., Zhang, J. and Rittenhouse, S.E. (1992) Activation-dependent changes in human platelet PECAM-1: phosphorylation, cytoskeletal association, and surface membrane redistribution, The Journal of cell biology, 119, 239-246. Newman, P.J. and Newman, D.K. (2003) Signal transduction pathways mediated by PECAM-1: new roles for an old molecule in platelet and vascular cell biology, Arteriosclerosis, thrombosis, and vascular biology, 23, 953-964. Newton, J.P., Buckley, C.D., Jones, E.Y. and Simmons, D.L. (1997) Residues on both faces of the first immunoglobulin fold contribute to homophilic binding sites of PECAM-1/CD31, The Journal of biological chemistry, 272, 20555-20563. O'Brien, C.D., Cao, G., Makrigiannakis, A. and DeLisser, H.M. (2004) Role of immunoreceptor tyrosine-based inhibitory motifs of PECAM-1 in PECAM-1-dependent cell migration, American journal of physiology, 287, C1103-1113. Okazaki, H., Ishizaka, N., Sakurai, T., Kurokawa, K., Goto, K., Kumada, M. and Takuwa, Y. (1993) Molecular cloning of a novel putative G protein-coupled receptor expressed in the cardiovascular system, Biochemical and biophysical research communications, 190, 1104-1109. Osawa, M., Masuda, M., Harada, N., Lopes, R.B. and Fujiwara, K. (1997) Tyrosine phosphorylation of platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31) in mechanically stimulated vascular endothelial cells, European journal of cell biology, 72, 229-237. Osawa, M., Masuda, M., Kusano, K. and Fujiwara, K. (2002) Evidence for a role of platelet endothelial cell adhesion molecule-1 in endothelial cell mechanosignal transduction: is it a mechanoresponsive molecule?, The Journal of cell biology, 158, 773-785. Pellegatta, F., Chierchia, S.L. and Zocchi, M.R. (1998) Functional association of platelet endothelial cell adhesion molecule-1 and phosphoinositide 3-kinase in human neutrophils, The Journal of biological chemistry, 273, 27768-27771. Pumphrey, N.J., Taylor, V., Freeman, S., Douglas, M.R., Bradfield, P.F., Young, S.P., Lord, J.M., Wakelam, M.J., Bird, I.N., Salmon, M. and Buckley, C.D. (1999) Differential association of cytoplasmic signalling molecules SHP-1, SHP-2, SHIP and phospholipase C-gamma1 with PECAM-1/CD31, FEBS letters, 450, 77-83. Rosen, H. and Goetzl, E.J. (2005) Sphingosine 1-phosphate and its receptors: an autocrine and paracrine network, Nature reviews, 5, 560-570. Sanchez, T. and Hla, T. (2004) Structural and functional characteristics of S1P receptors, Journal of cellular biochemistry, 92, 913-922. Schimmenti, L.A., Yan, H.C., Madri, J.A. and Albelda, S.M. (1992) Platelet endothelial cell adhesion molecule, PECAM-1, modulates cell migration, Journal of cellular physiology, 153, 417-428. Shida, D., Kitayama, J., Yamaguchi, H., Yamashita, H., Mori, K., Watanabe, T., Yatomi, Y. and Nagawa, H. (2004) Sphingosine 1-phosphate transactivates c-Met as well as epidermal growth factor receptor (EGFR) in human gastric cancer cells, FEBS letters, 577, 333-338. Shikata, Y., Birukov, K.G. and Garcia, J.G. (2003) S1P induces FA remodeling in human pulmonary endothelial cells: role of Rac, GIT1, FAK, and paxillin, J Appl Physiol, 94, 1193-1203. Su, W.H., Chen, H.I. and Jen, C.J. (2002) Differential movements of VE-cadherin and PECAM-1 during transmigration of polymorphonuclear leukocytes through human umbilical vein endothelium, Blood, 100, 3597-3603. Sukocheva, O., Wadham, C., Holmes, A., Albanese, N., Verrier, E., Feng, F., Bernal, A., Derian, C.K., Ullrich, A., Vadas, M.A. and Xia, P. (2006) Estrogen transactivates EGFR via the sphingosine 1-phosphate receptor Edg-3: the role of sphingosine kinase-1, The Journal of cell biology, 173, 301-310. Sun, J., Paddock, C., Shubert, J., Zhang, H.B., Amin, K., Newman, P.J. and Albelda, S.M. (2000) Contributions of the extracellular and cytoplasmic domains of platelet-endothelial cell adhesion molecule-1 (PECAM-1/CD31) in regulating cell-cell localization, Journal of cell science, 113 ( Pt 8), 1459-1469. Sun, J., Williams, J., Yan, H.C., Amin, K.M., Albelda, S.M. and DeLisser, H.M. (1996) Platelet endothelial cell adhesion molecule-1 (PECAM-1) homophilic adhesion is mediated by immunoglobulin-like domains 1 and 2 and depends on the cytoplasmic domain and the level of surface expression, The Journal of biological chemistry, 271, 18561-18570. Sun, Q.H., DeLisser, H.M., Zukowski, M.M., Paddock, C., Albelda, S.M. and Newman, P.J. (1996) Individually distinct Ig homology domains in PECAM-1 regulate homophilic binding and modulate receptor affinity, The Journal of biological chemistry, 271, 11090-11098. Tanimoto, T., Lungu, A.O. and Berk, B.C. (2004) Sphingosine 1-phosphate transactivates the platelet-derived growth factor beta receptor and epidermal growth factor receptor in vascular smooth muscle cells, Circulation research, 94, 1050-1058. Vaporciyan, A.A., DeLisser, H.M., Yan, H.C., Mendiguren, II, Thom, S.R., Jones, M.L., Ward, P.A. and Albelda, S.M. (1993) Involvement of platelet-endothelial cell adhesion molecule-1 in neutrophil recruitment in vivo, Science (New York, N.Y, 262, 1580-1582. Wang, F., Van Brocklyn, J.R., Hobson, J.P., Movafagh, S., Zukowska-Grojec, Z., Milstien, S. and Spiegel, S. (1999) Sphingosine 1-phosphate stimulates cell migration through a G(i)-coupled cell surface receptor. Potential involvement in angiogenesis, The Journal of biological chemistry, 274, 35343-35350. Wojciechowski, J.C. and Sarelius, I.H. (2005) Preferential binding of leukocytes to the endothelial junction region in venules in situ, Microcirculation, 12, 349-359. Wu, W.T., Chen, C.N., Lin, C.I., Chen, J.H. and Lee, H. (2005) Lysophospholipids enhance matrix metalloproteinase-2 expression in human endothelial cells, Endocrinology, 146, 3387-3400. Yamaguchi, F., Tokuda, M., Hatase, O. and Brenner, S. (1996) Molecular cloning of the novel human G protein-coupled receptor (GPCR) gene mapped on chromosome 9, Biochemical and biophysical research communications, 227, 608-614. Yatomi, Y. (2006) Sphingosine 1-phosphate in vascular biology: possible therapeutic strategies to control vascular diseases, Current pharmaceutical design, 12, 575-587. Yatomi, Y., Ohmori, T., Rile, G., Kazama, F., Okamoto, H., Sano, T., Satoh, K., Kume, S., Tigyi, G., Igarashi, Y. and Ozaki, Y. (2000) Sphingosine 1-phosphate as a major bioactive lysophospholipid that is released from platelets and interacts with endothelial cells, Blood, 96, 3431-3438. Yatomi, Y., Ozaki, Y., Ohmori, T. and Igarashi, Y. (2001) Sphingosine 1-phosphate: synthesis and release, Prostaglandins, 64, 107-122. Yatomi, Y., Ruan, F., Hakomori, S. and Igarashi, Y. (1995) Sphingosine-1-phosphate: a platelet-activating sphingolipid released from agonist-stimulated human platelets, Blood, 86, 193-202. Zhang, H., Desai, N.N., Olivera, A., Seki, T., Brooker, G. and Spiegel, S. (1991) Sphingosine-1-phosphate, a novel lipid, involved in cellular proliferation, The Journal of cell biology, 114, 155-167. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30346 | - |
dc.description.abstract | 鞘氨醇 1-磷酸鹽 (Sphingosine 1-phosphate; S1P) 是一個具有多種生物功能性的小型磷脂鹽類,S1P可以和隸屬於G型蛋白耦合受器的專屬膜上受器結合,傳遞訊息並藉此廣泛地調控各種細胞的行為。目前已知在內皮細胞中受到S1P影響的生物功能包括細胞增生、附著、移行、傷口癒合、細胞基質切割、管狀結構的生成、黏著因子表現、細胞素與趨化因子的合成與分泌等等。血小板/內皮細胞附著因子-1 (PECAM-1) 是一個細胞膜上的醣蛋白,主要表現在內皮細胞、血小板及一些血液細胞上。內皮細胞層上的PECAM-1分子大多聚集在細胞側邊的邊界上,這些PECAM-1會利用其胞外的結構和鄰近細胞的PECAM-1形成連結,藉此協助細胞之間的聯繫並控制細胞上下兩側的通透性。除了結構上的功能外,PECAM-1在胞內的分子結構組成包含了兩個特殊的免疫受體酪氨酸抑制基序 (immunoreceptor tyrosine inhibitory motifs; ITIM),當細胞遭遇到某些特定的外來刺激後,會使基序內含的酪氨酸產生磷酸化,磷酸化後的區位結構即會引發下游訊息分子的接合及活化,並引發後續的訊息傳遞及細胞生理活性的改變,然而在不同的來源細胞與外來刺激下,會分別經由不同種類的磷酸激酶來負責PECAM-1的磷酸化。目前已知PECAM-1在內皮細胞的移行、生存、管狀結構的生成、及免疫細胞對內皮細胞層的穿透過程中都扮演了很重要的角色。本篇研究目的在探討S1P引發人類臍靜脈內皮細胞上PECAM-1磷酸化的機制,首先使用免疫沉澱與西方墨點法來偵測蛋白質磷酸化態與總量的比例,觀察到S1P可以直接引發人類臍靜脈內皮細胞中PECAM-1的磷酸化,並確定這個磷酸化的現象與S1P的處理時間及處理濃度有關。在前處理了Gi與Src家族磷酸激酶的化學抑制劑後,可以確定這兩種訊息傳遞分子參與了這個磷酸化的過程。此外,我們利用西方墨點法與共軛焦顯微鏡驗證了在S1P處理後,位於側邊細胞邊界的活化態Src磷酸激酶呈現明顯的增加。為了確定是哪一個Src家族的磷酸激酶參與了PECAM-1磷酸化的過程,我們把目標鎖定在cSrc和Fyn這兩種細胞內主要的Src磷酸激酶成員上,將針對cSrc和Fyn的小型干擾核酸使用電破法轉殖入臍靜脈內皮細胞來截斷cSrc和Fyn的蛋白質表現,轉殖結束後持續培養細胞48小時再進行S1P的處理並比較不同的干擾核酸對S1P誘發PECAM-1磷酸化的效果,結果顯示cSrc和Fyn同時具有調控S1P所引發之PECAM-1磷酸化的能力。 | zh_TW |
dc.description.abstract | Sphingosine 1-phosphate (S1P) is a multifunctional bioactive lipid. Through a family of specific G protein-coupled receptors encoded by endothelial differentiation gene (edg), S1P regulates various cellular functions in different cell types. In endothelial cells, S1P has been demonstrated to modulate cell proliferation, adhesion, migration, wound healing, matix remodeling, tube formation, adhesion molecule expression, cytokine and chemokine releasing. Platelet/endothelial cell adhesion molecule-1 (PECAM-1) is a 130-kDa transmembrane glycoprotein which forms trans-homophilic bindings at cell-cell border. PECAM-1 performs both structural and signaling mediatory functions. Its cytoplasmic immunoreceptor tyrosine inhibitory motifs (ITIM) and tyrosine residues, when being phosphorylated, provide docking sites for various signaling molecules and modulate following cellular functions, such as cell migration, cell survival, tube formation, and transendothelial migration. However, the effect of S1P on endothelial PECAM-1 is still unknown, and the identification of the kinases responsible for PECAM-1 phosphorylation varies in different cell types upon different stimulations. In this study, we confirmed that S1P treatment induced PECAM-1 phosphorylation in human umbilical cord vein cells (HUVECs) by immunoprecipitation and following immunoblotting. The induction occurred immediately in less than three minutes. The induction could be block by pertussis toxin (PTx) and PP2 pretreatment, indicating the participation of Gi and Src family kinases. Results from immunoblotting and confocal microscopy proved that S1P also triggered the activation of Src family kinases at cell border. Finally, to identify the Src member responsible for PECAM-1 phosphorylation, specific siRNA was transfected to HUVECs. The obtained PECAM-1 phosphorylation ratio revealed that both cSrc and Fyn participated in this S1P-induced PECAM-1 phosphorylation. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T02:01:42Z (GMT). No. of bitstreams: 1 ntu-96-R94b41026-1.pdf: 1301537 bytes, checksum: d8b09e710a62d35efb62cc609bd1c376 (MD5) Previous issue date: 2007 | en |
dc.description.tableofcontents | 中文摘要 ………………………………………………………………Ⅰ
Abstract …………………………………………………………………Ⅲ Introduction …………………………………………………………….. 1 Generation and metabolism of sphingosine 1-phosphate ……………………………. 1 Spingosine 1-phosphate receptors …………………………………………………… 1 S1P effects on endothelial cells ……………………………………………………… 3 PECAM-1 expression and structure …………………………………………………. 3 PECAM-1 contains both structural and signal-transductional functions ……………. 4 PECAM-1 tyrosine phosphorylation regulates cellular functions …………………… 5 S1P effects on endothelial PECAM-1 ……………………………………………….. 7 Material and Methods …………………………………………………... 8 Reagents ………………………………………………………………...…………… 8 HUVEC Isolation ……………………………………………………………………. 9 Cell culture ………………………………………………………………………...… 9 S1P stimulation ………………………………………………………………………. 9 Immunoprecipitation ……………………………………………………………….. 10 Immunoblotting …………………………………………………………………….. 11 Immunofluorescence ……………………………………………………………….. 11 RNA interference and electroporation ……………………………………………… 12 Statistics analysis ………………………………………………………………….... 13 Results ………………………………………………………………… 14 S1P induced PECAM-1 phosphorylation through a time- and concentration- dependent manner …………………………………………………………………... 14 S1P-induced PECAM-1 phosphorylation was mediated through Gi and Src family kinases ……………………………………………………………………………… 15 S1P induced cSrc and Fyn activation at cell border ………………………………... 16 S1P induced PECAM-1 phosphorylation through both cSrc and Fyn …………...… 17 Discussion ……………………………………………………………... 19 References …………………………………………………………….. 25 Figures ………………………………………………………………… 34 | |
dc.language.iso | en | |
dc.title | 鞘氨醇 1-磷酸鹽引發人類內皮細胞表面之血小板/內皮細胞附著因子-1磷酸化機制之研究 | zh_TW |
dc.title | Studies of the Mechanisms of Sphingosine 1-phosphate Induces Platelet/Endothelial Cell Adhesion Molecule-1
Phosphorylation in Human Endothelial Cells | en |
dc.type | Thesis | |
dc.date.schoolyear | 95-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳俊宏,黃偉邦,沈湯龍 | |
dc.subject.keyword | 鞘氨醇 1-磷酸鹽,內皮細胞,血小板/內皮細胞附著因子,磷酸化, | zh_TW |
dc.subject.keyword | sphingosine 1-phosphate,endothelial cells,PECAM-1,phosphorylation,Src family kinases, | en |
dc.relation.page | 47 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2007-07-09 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 動物學研究研究所 | zh_TW |
顯示於系所單位: | 動物學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-96-1.pdf 目前未授權公開取用 | 1.27 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。