(3.236.175.108) 您好!臺灣時間:2021/02/27 06:47
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:蔡世傑
研究生(外文):Shi-Jeih Tsai
論文名稱:GDNF/RET引起MDCK細胞移行之訊息傳遞研究
論文名稱(外文):Signal Mechanisms of GDNF/RET-Induced Migration in MDCK Cells
指導教授:湯銘哲湯銘哲引用關係
指導教授(外文):Ming-Jer Tang
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:1999
畢業學年度:87
語文別:英文
論文頁數:49
中文關鍵詞:重組誘發轉型膠質細胞衍生性神經營養因子膠質細胞衍生性神經營養因子甲型受體細胞移行磷脂醯肌醇-3激化脢
外文關鍵詞:RETGDNFGDNFR-alphaCell MigrationPhosphatidylinositol 3-kinase
相關次數:
  • 被引用被引用:0
  • 點閱點閱:178
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
RET(Rearrangement-induced Transformation)是一種受體酪氨酸激化脢(Receptor tyrosine kinase),目前已知與腎臟及周邊神經系統之發育有關。活化RET需要其他兩種分子同時存在:膠質細胞衍生性神經營養因子(Glial cell line-Derived Neurotrophic Factor,GDNF)及GDNFR-alpha,一種醣磷脂醯肌醇連結之細胞表面受體(Glycosyl phosphatidylinositol-linked cell surface receptor)。為了研究RET在細胞內之訊息傳遞機制,我們建立了一個將RET大量表現在MDCK細胞株的系統;在此系統中已經證實了RET之活化與細胞移行(Migration)、散生(Scattering)、以及細胞伸出偽足(Filopodia and lamellipodia)等現象有關。在此,我們又建立了一些表現突變RET之MDCK細胞株,如正常之RET(Ret9,Ret2),激化脢失去功能之RET(Kinase mutant, km),及一些將不同位置之酪氨酸(Tyrosine,Y)置換為苯丙氨酸(Phenylalanine,F)之RET如M1062(Y1062F)、M1090(Y1090F)、M2(Y1090F and Y1096F)、M708(Y660F, Y687F, Y826F, Y1015F, Y1029F, Y1062F, and Y1096F)及M816(Y660F, Y687F, Y826F, Y1015F, Y1029F, Y1062F, Y1090F, and Y1096F)。在這些表現不同突變RET之細胞中,Ret9細胞經GDNF/GDNFR-處理可促進其細胞增生、移行能力及散生增加。而m和M816在GDNF/GDNFR-alpha的刺激下並沒有任何型態、移行及增生方面的改變。有趣的是,GDNF/GDNFR-alpha會使M708細胞散生及移行增加,但不會促進生長。另外,M1062及M2兩株細胞的細胞移行能力較正常RET之細胞如Ret9,Ret2等之移行來的低,但M1090細胞之移行能力則非常顯著,因此我推測Tyr1096可分別正向調控Tyr1062及負向調控Tyr1090這兩個酪氨酸送出影響細胞移行的訊息。我們也發現了RET的活化可以引起磷脂醯肌醇-3激化脢(Phosphatidylinositol 3-Kinase ,PI3-kinase)的活性。而用磷脂醯肌醇-3激化脢之抑制劑LY294002來處理Ret9、M708及M1090三種細胞則可抑制GDNF/RET所引發之細胞移行。綜合上述之結果,我們認為由RET之Tyr1062、Tyr1090及Tyr1096所下傳的訊息與細胞移行有關,且這些引起細胞移行的訊息可能是透過磷脂醯肌醇-3激化脢所調控的。

RET (Rearrangement-induced Transformation) is a receptor tyrosine kinase involved in the development of kidney and certain parts of the peripheral nervous system. The activation of RET requires GDNF (Glial cell line-Derived Neurotrophic Factor) and GDNFR-, a glycosyl phosphatidylinositol-linked cell surface receptor. It has been demonstrated that activation of RET induces cell migration, scattering and filopodia and lamellipodia formation in RET overexpressing MDCK cells. To investigate the signal pathway mechanisms of RET activation, we employed MDCK cells stably transfected with wild type RET (Ret9, Ret2), kinase dead mutant (Km), and various mutants in which different tyrosine residues (Y) were substituted with phenylalanine (F) including M1090 (Y1090F), M2 (Y1090F and Y1096F), M1062 (Y1062F), M708 (Y660F, Y687F, Y826F, Y1015F, Y1029F, Y1062F, and Y1096F) and M816 (Y660F, Y687F, Y826F, Y1015F, Y1029F, Y1062F, Y1090F, and Y1096F). Among these transfectants, neither Km nor M816 cells showed any morphological or growth effects upon RET activation by addition of GDNF/GDNFR-alpha. Interestingly, M708 exhibited marked increase in migration and scattering, but no change in proliferation in response to GDNF/GDNFR-alpha indicating the role of Tyr1090 in cell migration. In addition, M2 and M1062 exhibited decreased migration capacity whereas M1090 migrated significantly, suggesting that Tyr1062 plays roles in cell migration and Tyr1096 may serve as a positive regulator for Tyr1062 and inhibitor for Tyr1090 in GDNF/RET-induced cell motility. We also found that GDNF/RET interactions resulted in enhanced phosphatidylinositol 3-kinase (PI3-kinase) activity. LY294002, a PI3-kinase inhibitor, blocked GDNF- induced migration effects in Ret9 and M708 cells. Taken together, Tyr1062, Tyr1090 and Tyr1096 of RET may mediate GDNF/RET-induced cell migration through activation of PI3-kinase.

中文摘要.....................1
ABSTRACT.....................3
INTRODUCTION.................5
MATERIALS AND METHODS........12
RESULTS......................21
DISCUSSION...................26
REFERENCES...................30
FIGURES......................37
誌謝.........................49

Alberti, L., Borrello, M.G., Ghizzoni, S., Torriti, F., Rizzetti, M. G., and Pierotti, M. A. Grb2 binding to the different isoforms of Ret tyrosine kinase. Oncogene 17:1079-1087, 1998.
Asai, N., Murakami, H., Iwashita, T., and Takahashi, M. A mutation at tyrosine 1062 in MEN2A-Ret and MEN2B-Ret impairs their transforming activity and association with Shc adaptor proteins. J. Biol. Chem. 271:17644-17649, 1996.
Cantley, L. C., Auger, K. R., Carpenter, C., Duckworth, B., Graziani, A., and kapeller, R. Oncogenes and signal transduction. Cell 64:281-302, 1991.
Chiariello, M., Visconti, R., Carlomagno, F., Melillo, R. O., Bucci, C., Franciscis, V. D., Fox, G. M., Jing, S., Coso, O. A., Gutkind, J. S., Fusco, A., and Santoro, M. Signalling of the Ret receptor tyrosine kinase through the c-Jun NH2-terminal protein kinases (JNKs): evidence for a divergence of the ERKs and JNKs pathways induced by Ret. Oncogene 16: 2435-2445, 1998.
Cooper, J. A. The role of actin polymerization in cell motility. Annu. Rev. Physiol. 53:585-605, 1991.
Donis-keller, H., Dou, S., Chi, D., Carlson, K. M., Toshima, K., Jackson, C. E., Wells, S. A., Goodfellow, P. J., and Donis-keller, H. Single missense mutation in the tyrosine kinase catalytic domain of the RET protooncogene is associated with multiple endocrine neoplasia type 2B. Proc. Natl. Acad. Sci. U.S.A. 91:1579-1583, 1994
Durick, K., Gill, G. N., and Taylor, S. S. Shc and engima are both required for mitogenic signaling by Ret/ptc2. Mol. Cell Biol. 18:2298-1308, 1998.
Durick, K., Wu, R. Y., Gill, G. N., and Taylor, S. S. Mitogenic signalling by Ret/ptc2 requires association with enigma via a LIM domain. J. Biol. Chem. 271:12691-12694, 1996.
Edery, P., Lyonnet, S., Mulligan, L. M., Pelet, A., Dow, E., Abel, L., Holder, S., Ninoul-fekete, C., Ponder, B. A. J., and Munnich, A. Mutations of the RET proto-oncogene in Hirschsprung's disease. Nature 367:378-380, 1994.
Hall, A. Rho GTPases and the actin cytoskeleton. Science 279:509-514, 1998.
Hellmich, H. L., Kos, L., Cho, E. S., Mahon, K. A. and Zimmer, A. Embryonic expression of glial cell-line derived neurotrophic factor (GDNF) suggests multiple developmental roles in neural differentiation and epithelial-mesenchymal interactions. Mech. Dev. 54:95-106, 1996.
Hofstra, R. M., Landsvater, R. M., Ceccherini, I., Stulp, R. P., Stelwagen, T., Lips, C. J. M., and Buys, C. H. C. M. A mutation in the RET proto-oncogene associated with multiple endocrine neoplasia type 2B and sporadic medullary thyroid carcinoma. Nature 367:375-376, 1994.
Jing, S., Wen, D., Yu, Y., Holst, P. L., Luo, Y., Fang, M., Tamir, R., Antonio, L., Hu, Z., Cupples, R., Louis, J.-C., Hu, S., Altrock, B. W. and Fox, G. M. GDNF-induced activation of the Ret protein tyrosine kinase is mediated by GDNFR-, a novel receptor for GDNF. Cell 85:1113-1124, 1996.
Keely, P. J., Westwick, J. K., Whitehead, I. P., Der, C. J., and Parise, L. V. Cdc42 and rac1 induce integrin-mediated cell motility and invasiveness through PI(3)K. Nature 390:632-636, 1997.
Khwaja, A., Lehmann, K., Marte, B. M., and Downward, J. Phosphoinositide 3-kinase induces scattering and tubulogenesis in epithelial cells through a novel pathway. J. Biol. Chem. 273:18793-18801,1998.
Konati, K., Yonezawa, K., Hara, K., Ueda, H., Kitamura, Y., Sakaue, H., Ando, A., Chavaneau, A., Calas, B., grigorescu, F., Involvement of phosphoinositide 3-kinase in insulin or IGF-1 induced membrane ruffling. EMBO J. 13:2313-2321, 1994.
Lechner, M. S. and Dressler, G. R., The molecular basis of embryonic kidney development. Mech. Dev. 62: 105-120, 1997.
Lin, L.-F. H., Doherty, D. H., Lile. J. D., bektesh, S., and Collins, F. GDNF: a glial cell line-derived neurotrophic factor for midbrain dopaminergic neurons. Science 260:1130-1132, 1993.
Liu, X., Vega, Q. C., Decker, R. A., Pandey, A., Worby, C. A., and Dixon J. C. Oncogenic RET receptors display different autophosphorylation sites and substrate binding specificites. J. Biol. Chem. 271: 5309-5312, 1996.
Moore, M. W., Klein, R. D., Farinas, I., Sauer, H., Armanini, M., Phillips, H., Reichardt, L. F., Ryans, A. M., Carver-moore, K., and Rosenthal, A. renal and neuronal abnormalities in mice lacking GDNF. Nature 382:76-79, 1996.
Mulligan, L. M., Kwok, J. B. J., Healey, C. S., Elsdon, M. J., Eng, C., Gardner, E., Love, D. R., Mole, S. E., Moore. J. K., Papi, L., Ponder, M. A., Telenius, H., Tunnacliffe, A., and Ponder, B. A. J. Germ-line mutations of the RET proto-oncogene in multiple endocrine neoplasia type 2A. Nature 363:458-460, 1993.
Nobes, C. D., Hawkins, P., Stephens, L., and Hall, A. Activation of the small GTP-binding proteins rho and rac by growth factor receptors. J. cell Sci. 108:225-233, 1995.
O’Rourke, D. A., Sakurai, H., Spokes, K., Kjelsberg, C., Takahashi, M., Nigam, S., and Cantley, L. G. Expression of c-ret promotes morphogenesis and cell survival in mIMCD-3 cells. Am. J. Physiol. 276:F581-F588, 1999.
Pandey, A., Duan, H., Fiore, P. P. D., and Dixit, V. M. The Ret receptor protein tyrosine kinase associates with the SH2-containing adapter protein Grb2. J. Biol. Chem.270: 21461-21463, 1995.
Pelicci, G., Lanfrancone, L., Grignani, F., McGlade, J., Cavallo, F., Forni, G., Nicoletti, I., Pawson, T., and Pelicci, P. G. A novel transforming protein (SHC) with an SH2 domain is implicated in mitogenic signal transduction. Cell 70:93-104, 1992.
Pichel, J. G., Shen, L., Sheng, H. Z., Granholm, A.-C. Drago, J., Grinberg, A., Lee, E. J., Huang, S. P., Saarma, M., Hoffer, B. J., Sariola, H. and Westphal, H. Defects in enteric innervation and kidney development in mice lacking GDNF. Nature 382:73-76, 1996.
Ridley, A. J., Comoglio, P. M., and Hall, A. Regulation of scatter factor/hepatocyte growth factor responses by Ras, Rac, and Rho in MDCK cells. Mol. Cell. Biol. 15:1110-1122, 1995.
Rodriguez-Viciana, P., Warne, P. H., Khwaja, A., Marte, B. M., Pappin, D., Das, P., Waterfield, M. D., Ridley, A.,and Downward, J. Role of phosphoinositide 3-OH kinase in cell transformation and control of the actin cytoskeleton by Ras. Cell 89:457-467, 1997.
Romeo, G., Ronchetto, P., Luo, Y., Barone, V., Seri, M., Ceccerini, L., Pasini, B., Bocciardi, R., lerone, M. Kaariainen, H., and Martucciello, G. Point mutations affecting the tyrosine kinase domain of the RET proto-oncogene in Hirschsprung's disease. Nature 367:377-378, 1994.
Sanchez, M. P., Silos-Santiago, I., Frisen, J., He, B., Lira, S. A. and Barbacid, M. Renal agenesis and the absence of enteric neurons in mice lacking GDNF. Nature 382:70-73, 1996.
Santoro, M., carlomagno, F., Romano, A., Bottaro, D. P., Dathan, N. A., Grieco, M., Fusco, A., Vecchio, G., Matoskova, B., kraus, M. H., and Di fiore, P. P. Activation of RET as a dominant transforming gene by germline mutations of MEN2A and MEN2B. Science 267:381-383, 1995.
Schuchardt, A., D’Agati, V., Larsson-Blomberg, L., Costantini, F., and Pachnis, V. Defects in the kidney and enteric nervous system of mice lacking the tyrosine kinase receptor Ret. Nature 367:380-383, 1994
Songyang, Z., Shoelson, S, E., Chaudhuri, M., Gish, G., Pawson, T., Haser, W. G., King, F., Roberts, T., Ratnofsky, S., lechleider, R. J., SH2 domains recognize specific phosphopeptide sequences. Cell 72:767-778, 1993.
Takahashi M., Ritz J., and Cooper G. M. Activation of a novel human transforming gene, ret, by DNA rearrangement. Cell 42:581-588, 1985.
Tang, M. J. Worby, D., Sanicola, M., and Dressler, G. R. The RET-Glial cell-derived neurotrophic factor (GDNF) pathway stimulates migration and chemoattraction of epithelial cells. J. Cell Biol. 142:1337-1345,1998.
Van Weering, D. H. J., and J. L. Bos. Glial cell line-derived neurotrophic factor induces ret-mediated lamellipodia formation. J. Biol. Chem. 272:249-254, 1997.
Vanhaesebroeck, B., Leevers, S. J., Panayotou, G., and Waterfield, M. D. Phosphoinositide 3-kinases: a conserved family of signal transducers. Trends Biochem. Sci. 21:181-185, 1996.
Vega, G. C., Worby, C. A., Lechner, M. S., Dixon, J. E., and Dressler, G. R. Glial cell line-derived neurotrophic factor activates RET and promotes kidney morphogenesis. Proc. Natl. Acad. Sci. U.S.A. 93:10657-10661, 1996.
Wennstrom, S., Hawkins, P., Cooke, F., hara, K., Yonezawa, K., kasuga, M., Jackson, T., Claesson-Welsh, L., and Stephens, L. Activation of phosphoinositide 3-kinase is required for PDGF-stimulated membrane ruffling. Curr. Biol. 4:385-393, 1994.

QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
系統版面圖檔 系統版面圖檔