臺灣博碩士論文加值系統

鉀氯離子共同運輸蛋白是調控人體滲透壓平衡的主要決定因子之一。本實驗室先前一系列研究已發現鉀氯離子共同運輸蛋白在癌症進程上扮演許多重要角色。而本研究目的則是釐清鉀氯離子共同運輸蛋白在腫瘤生物學中的重要角色及調控機制。本研究包含四個部份。(一) 第四型鉀氯離子共同運輸蛋白(KCC4)表現量和癌細胞轉移能力的相關性。從雷射顯微切割技術加上即時定量系統，以及應用免疫螢光染色實驗所得到的結果，都證實轉移癌組織會表現大量的第四型鉀氯離子共同運輸蛋白，而有利於癌細胞的侵蝕性爬行及轉移。此外，第四型鉀氯離子共同運輸蛋白在轉移性癌組織中，和第一型類胰島素生長因子(IGF-1)及上皮細胞生長因子(EGF)有一共同表現的趨勢，顯示第四型鉀氯離子共同運輸蛋白在癌細胞的功能及表現可受到這二種生長因子的調節。(二) 探討第四型鉀氯離子共同運輸蛋白在癌細胞侵蝕的重要角色及其獨特調節機轉。利用卵巢癌及肺腺癌細胞株實驗模式中及配光學影像技術發現，第一型類胰島素生長因子及上皮細胞生長因子會促進第四型鉀氯離子共同運輸蛋白從細胞內胞器(如：內質網及高爾基氏體)運輸到侵蝕性癌細胞之前端細胞膜上；第四型鉀氯離子共同運輸蛋白於此處和細胞骨架結合蛋白(ezrin)相互作用以調控細胞骨架結構，進而促進癌細胞的侵蝕及轉移能力。(三) 以果蠅為活體實驗模式研究鉀氯離子共同運輸蛋白在上皮細胞發展的新穎功能。從基因學及分子生物學實驗得到的實驗結果顯示，果蠅的鉀氯離子共同運輸蛋白為胚胎發育過程的必需因子，而在神經系統活性及體內鹽份恆定方面扮演重要角色。此外，果蠅的鉀氯離子共同運輸蛋白對於果蠅的翅膀上皮細胞發育也很重要，主要藉由和上皮細胞中的細胞骨架蛋白(actin)結合而調控翅膀觸毛的生長。此研究結果可支持未來利用果?為活體模式進一步探討鉀氯離子共同運輸蛋白在腫瘤生物學上的其它新穎功能。(四) 探討鉀氯離子共同運輸對於癌細胞爬行的獨特調節機轉。我們探討鉀氯離子共同運輸活性是否對於上皮細胞生長因子引起的細胞骨架及細胞黏附(focal adhesions)的動態平衡有所影響，進而調控癌細胞的爬行活性。上皮細胞生長因子可能會調節KCC4和beta 1 integrin、ezrin、及focal adhesion kinase (FAK)之間的動態交互作用，進而影響爬行性癌細胞中細胞骨架重整過程。總結以上，此研究對於了解鉀氯離子共同運輸蛋白在癌細胞的侵蝕能力之角色及重要性將有很大的進展，並且提供基礎研究證據支持鉀氯離子共同運輸蛋白未來應用於臨床上癌症的診斷及治療。

The KCl cotransporter (KCC) is a major determinant of osmotic homeostasis and plays an important role in cancer development and progression. My thesis focuses on the emerging role for KCl cotransport in tumor biology and the novel mechanisms by which KCl cotransport regulates cancer malignant behaviors. My thesis includes four parts. (1) KCC4 expression is associated with cancer metastasis and clinical outcome. This part of study aims to investigate the contribution of individual KCC isoforms in cancer metastasis using cervical cancer and ovarian cancer as the model. The results indicate that metastatic cancer tissues express abundant KCC4 which benefits cancer cells in invasiveness. In the metastatic cancer tissues, KCC4 colocalizes with IGF-1 or EGF, indicating a likely in vivo stimulation of KCC4 function by growth factors. (2) Membrane trafficking of KCC4 is important for cancer cell invasion. Here I test the hypothesis that the regulation of specific KCC activation in cancer cells is a dynamic process which can be significantly upregulated by IGF-1 or EGF. The results indicate that IGF-1 and EGF stimulate the membrane recruitment of KCC4, in which KCC4 interacts with an actin-binding protein, ezrin, at lamellipodia. In addition to ion transport, KCC4 can function as a membrane scaffold to facilitate the modulation of cytoskeletal reorganization that is required for the invasive migration of cancer cells. (3) KCl cotransporter is an evolutionarily conserved assembly factor for actin-containing cellular protrusions. The novel functions of KCC in epithelial development were investigated using Drosophila melanogaster as a model. With the generation of KCC null mutants, it is suggested that fly KCC has essential role in the embryonic development and in the regulation of neural activity and salt homeostasis. Moreover, KCC is present in actin-bundle containing cellular protrusions of wing hairs and may play a role in the development of wing epithelium. (4) KCl cotransport is important for actin reorganization and focal adhesion dynamics during cancer cell migration. Here I test the hypothesis that KCl cotransport regulates cancer cell invasive migration is via the modulation of actin cytoskeleton and focal adhesions. The results suggest that KCl cotransport is necessary for actin reorganization and focal adhesion dynamics during cancer cell migration. Taken together, the results of my thesis provide the rationale for the clinical application of KCC as the therapeutic target and the prognostic biomarkers of metastatic cancers.

Abstract ………………………………………………………………………… 02 Chapter 1. Introduction ……………………………………………………… 06
1.1. Introduction of KCl cotransport family ……………… 07
1.1.1. KCC1 ………………………………………………………………… 09
1.1.2. KCC2 ………………………………………………………………… 10
1.1.3. KCC3 ………………………………………………………………… 11
1.1.4. KCC4 ………………………………………………………………… 13
1.2. The important role of KCl cotransport in tumor progression …………… 15
1.2.1. KCl cotransport activity affects tumor invasion and proliferation … 15
1.2.2. The important role of KCC3 in epithelial-mesenchymal transition (EMT) ………………………………………………………………17
1.3. Specific aims of the studies ………… 20
Chapter 2. KCC4 expression is associated with cancer metastasis and clinical outcome……………………………………………………………………22
Chapter 3. Membrane trafficking of KCC4 is important for cancer cell invasion …………………………………… 47
Chapter 4. KCl cotransporter is an evolutionarily conserved assembly factor for actin-containing cellular protrusions ……………………… 75
Chapter 5. KCl cotransport is important for actin reorganization and focal adhesion dynamics during cancer cell migration …………………………93
Chapter 6. Conclusions and prospects ……………………………… 112
References ……………………………………………………………124
Publication list ……………… 135

Adragna NC, Di Fulvio M, Lauf PK. (2004) Regulation of K-Cl cotransport: from function to genes. J Membr Biol 201:109-137.
Albini A, Allavena G, Melchiori A, Giancotti F, Richter H, Comoglio PM, Parodi S, Martin GR, Tarone G. (1987) Chemotaxis of 3T3 and SV3T3 cells to fibronectin is mediated through the cell-attachment site in fibronectin and a fibronectin cell surface receptor. J Cell Biol 105:1867-1872.
Ashbumer M (1989) Drosophila: a laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
Aznavorian S, Murphy AN, Stetler-Stevenson WG Liotta LA. (1993) Molecular aspects of tumour cell invasion and metastasis. Cancer 71:1368–1383.
Boettger T, H?bner CA, Maier H, Rust MB, Beck FX, Jentsch TJ. (2002) Deafness and renal tubular acidosis in mice lacking the K-Cl co-transporter Kcc4. Nature 416:874-878.
Boettger T, Rust MB, Maier H, Seidenbecher T, Schweizer M, Keating DJ, Faulhaber J, Ehmke H, Pfeffer C, Scheel O, Lemcke B, Horst J, Leuwer R, Pape HC, V?lkl H, H?bner CA, Jentsch TJ. (2003) Loss of K-Cl co-transporter KCC3 causes deafness, neurodegeneration and reduced seizure threshold. EMBO J 22:5422-5434.
Brand FX, Ravanel N, Gauchez AS, Pasquier D, Payan R, Fagret D, Mousseau M. (2006) Prospect for anti-HER2 receptor therapy in breast cancer. Anticancer Res 26:463–470.
Brand AH, Perrimon N. (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118:401-415.
Bretscher A, Edwards K, Fehon RG. (2002) ERM proteins and merlin: integrators at the cell cortex. Nat Rev Mol Cell Biol 3:586-899.
Brugnara C, Bunn HF, Tosteson DC. (1986) Regulation of erythrocyte cation and water content in sickle cell anemia. Science 232:388-390.
Byun N, Delpire E. Axonal and periaxonal swelling precede peripheral neurodegeneration in KCC3 knockout mice. Neurobiol Dis 28:39-51.
Cannistra SA, Ottensmeier C, Niloff J, Orta B, DiCarlo (1995) J. Expression and function of ?1 and ?v?3 integrins in ovarian cancer. Gynecol Oncol 58:216-225.
Casula S, Shmukler BE, Wilhelm S, Stuart-Tilley AK, Su W, Chernova MN, Brugnara C, Alper SL. (2001) A dominant negative mutant of the KCC1 K-Cl cotransporter: both N- and C-terminal cytoplasmic domains are required for K-Cl cotransport activity. J Biol Chem 276:41870-41878.
Chee KS, Kistler J, Donaldson PJ. (2006) Roles for KCC transporters in the maintenance of lens transparency. Invest Ophthalmol Vis Sci 47:673-682.
Chen YF, Chou CY, Wilkins RJ, Ellory JC, Mount DB, Shen MR. (2009) Motor protein-dependent membrane trafficking of KCl cotransporter-4 is important for cancer cell invasion. Cancer Res 69:8585-8593.
Chiang Y, Chou CY, Hsu KF, Huang YF, Shen MR. (2008) EGF upregulates Na+/H+ exchanger NHE1 by post-translational regulation that is important for cervical cancer cell invasiveness. J Cell Physiol 214:810-819.
Christofori G. (2006) New signals from the invasive front. Nature 441:444–450.
del Pozo MA, Alderson NB, Kiosses WB, Chiang HH, Anderson RGW, Schwartz MA. (2004) Integrins regulate Rac targeting by internalization of membrane domains. Science 303:839-842.
Delpire E, Mount DB. (2002) Human and murine phenotypes associated with defects in cation-chloride cotransport. Annu Rev Physiol 64:803–843.
Denker SP, Barber DL. (2002) Cell migration requires both ion translocation and cytoskeletal anchoring by the Na-H exchanger NHE1. J Cell Biol 159:1087-1096.
Denker SP, Huang DC, Orlowski J, Furthmayr H, Barber DL. (2000) Direct binding of the Na--H exchanger NHE1 to ERM proteins regulates the cortical cytoskeleton and cell shape independently of H(+) translocation. Mol Cell 6:1425-1436.
Deryugina EI, Quigley JP. (2006) Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev 25:9-34.
Dunham PB, Stewart GW, Ellory JC. (1980) Chloride-activated passive potassium transport in human erythrocytes. Proc Natl Acad Sci USA 77:1711-1715.
Ezratty EJ, Partridge MA, Gundersen GG. (2005) Microtubule-induced focal adhesion disassembly is mediated by dynamin and focal adhesion kinase. Nat Cell Biol 7:581–590.
Flatman PW, Adragna NC, Lauf PK. (1996) Role of protein kinases in regulating sheep erythrocyte K-Cl cotransport. Am J Physiol 271:C255-C263.
Fehrmann RS, Li XY, van der Zee AG, de Jong S, Te Meerman GJ, de Vries EG, Crijns AP. (2007) Profiling studies in ovarian cancer: a review. Oncologist 12:960-966.
Friedl P, Wolf K. (2003) Tumour-cell invasion and migration: diversity and escape mechanisms. Nat Rev Cancer 3:362-374.
Fehon RG, McClatchey AI, Bretscher A. (2010) Organizing the cell cortex: the role of ERM proteins. Nat Rev Mol Cell Biol.11:276-87.
Filippov V, Aimanova K, Gill SS. (2003) Expression of an Aedes aegypti cation-chloride cotransporter and its Drosophila homologoues. Insect Mol Biol 12:319–331.
F?rstenberger G, Senn HJ. (2002) Insulin-like growth factors and cancer. Lancet Oncol 3:298-302.
Gaus K, Le Lay S, Balasubramanian N, Schwartz MA. (2006) Integrin-mediated adhesion regulates membrane order. J Cell Biol 174:725-734.
Garz?n-Muvdi T, Pacheco-Alvarez D, Gagnon KB, V?zquez N, Ponce-Coria J, Moreno E, Delpire E, Gamba G. (2007) WNK4 kinase is a negative regulator of K+-Cl- cotransporters. Am J Physiol Renal Physiol 292:F1197-F1207.
Gillen CM, Brill S, Payne JA, Forbush B 3rd. (1996) Molecular cloning and functional expression of the K-Cl cotransporter from rabbit, rat, and human. A new member of the cation-chloride cotransporter family. J Biol Chem 271:16237-16244.
Guan JL. (1997) Role of focal adhesion kinase in integrin signaling. Int J Biochem Cell Biol 29:1085–96.
Hanahan D, Weinberg RA. (2000) The hallmarks of cancer. Cell 100:57–70.
Hekmat-Scafe DS, Lundy MY, Ranga R, Tanouye MA. (2006) Mutations in the K+/Cl- cotransporter gene kazachoc (kcc) increase seizure susceptibility in Drosophila. J Neurosci 26:8943-54.
Hekmat-Scafe DS, Mercado A, Fajilan AA, Lee AW, Hsu R, Mount DB, Tanouye MA. (2006) Seizure sensitivity is ameliorated by targeted expression of K+-Cl- cotransporter function in the mushroom body of the Drosophila brain. Genetics. 184:171-83.
Hiki K, D'Andrea RJ, Furze J, Crawford J, Woollatt E, Sutherland GR, Vadas MA, Gamble JR. (1999) Cloning, characterization, and chromosomal location of a novel human K+-Cl- cotransporter. J Biol Chem 274:10661-10667.
Holtzman EJ, Kumar S, Faaland CA, Warner F, Logue PJ, Erickson SJ, Ricken G, Waldman J, Kumar S, Dunham PB. (1998) Cloning, characterization, and gene organization of K-Cl cotransporter from pig and human kidney and C. elegans. Am J Physiol 275:F550-F564.
Howard HC, Mount DB, Rochefort D, Byun N, Dupre N, Lu J, Fan X, Song L, Riviere JB, Prevost C, Horst J, Simonati A, Lemcke B, Welch R, England R, Zhan FQ, Mercado A, Siesser WB, George AL Jr, McDonald MP, Bouchard JP, Mathieu J, Delpire E, Rouleau GA. (2002) The K-Cl cotransporter KCC3 is mutant in a severe peripheral neuropathy associated with agenesis of the corpus callosum. Nat Genet 32:384–392.
Hsu YM, Chou CY, Chen HH, Lee WY, Chen YF, Lin PW, Alper SL, Ellory JC, Shen MR. (2007a) IGF-1 upregulates electroneutral K-Cl cotransporter KCC3 and KCC4 which are differentially required for breast cancer cell proliferation and invasiveness. J Cell Physiol 210:626-636.
Hsu YM, Chen YF, Chou CY, Tang MJ, Chen JH, Wilkins RJ, Ellory JC, Shen MR. (2007b) KCl cotransporter-3 down-regulates E-cadherin/beta-catenin complex to promote epithelial-mesenchymal transition. Cancer Res 67:11064-11073.
H?bner CA, Stein V, Hermans-Borgmeyer I, Meyer T, Ballanyi K, Jentsch TJ. (2001) Disruption of KCC2 reveals an essential role of K-Cl cotransport already in early synaptic inhibition. Neuron 30:515-524.
Ilic D, Damsky CH, Yamamoto T. (1997) Focal adhesion kinase: at the crossroads of signal transduction. J Cell Sci 110:401–407.
Insall RH, Machesky LM. (2009) Actin dynamics at the leading edge: from simple machinery to complex networks. Dev Cell 17:310-322.
Jennings ML, al-Rohil N. (1990) Kinetics of activation and inactivation of swelling-stimulated K+/Cl- transport. The volume-sensitive parameter is the rate constant for inactivation. J Gen Physiol 95:1021-1040.
Joiner CH, Rettig RK, Jiang M, Risinger M, Franco RS. (2007) Urea stimulation of KCl cotransport induces abnormal volume reduction in sickle reticulocytes. Blood 109:1728-1735.
Kahle KT, Rinehart J, de Los Heros P, Louvi A, Meade P, Vazquez N, Hebert SC, Gamba G, Gimenez I, Lifton RP. (2005)　WNK3 modulates transport of Cl- in and out of cells: implications for control of cell volume and neuronal excitability. Proc Natl Acad Sci USA 102:16783-16788.
Karadsheh MF, Byun N, Mount DB, Delpire E. (2004) Localization of the KCC4 potassium-chloride cotransporter in the nervous system. Neuroscience 123:381-391.
Kim B, van Golen CM, Feldman EL. (2004) Insulin-like growth factor-I signaling in human neuroblastoma cells. Oncogene 23:130-141.
Lang F, Busch GL, Ritter M, V?lkl H, Waldegger S, Gulbins E, H?ussinger D. (1998) Functional significance of cell volume regulatory mechanisms. Physiol Rev 78:247-306.
Lauf PK, Bauer J, Adragna NC, Fujise H, Zade-Oppen AMM, Ryu KH, Delpire E. (1992) Erythrocyte K-Cl cotransport: properties and regulation. Am J Physiol 263:C917-C932.
Lauf PK, Theg BE. (1980) A chloride dependent K+ flux induced by N-ethylmaleimide in genetically low K+ sheep and goat erythrocytes. Biochem Biophys Res Commun 92:1422-1428.
Lauf PK. (1982) Evidence for chloride-dependent potassium and water transport induced by hyposmotic stress in erythrocytes of the marine teleost, Opsanus tau. J Comp Physiol 146:9-16.
Lauf PK, Adragna NC. (2000) K-Cl cotransport: properties and molecular mechanism. Cell Physiol Biochem 10:341-354.
Lee JM, Dedhar S, Kalluri R, Thompson EW. (2006) The epithelial-mesenchymal transition: new insights in signaling, development, and disease. J Cell Biol 172:973–981.
Lee MY, Chou CY, Tang MJ, Shen MR. (2008) Epithelial-mesenchymal transition in cervical cancer: correlation with tumor progression, epidermal growth factor receptor overexpression, and snail up-regulation. Clin Cancer Res 14:4743-4750.
Lew VL, Bookchin RM. (2005) Ion transport pathology in the mechanism of sickle cell dehydration. Physiol Rev 85:179-200.
Li H, Khirug S, Cai C, Ludwig A, Blaesse P, Kolikova J, Afzalov R, Coleman SK, Lauri S, Airaksinen MS, Kein?nen K, Khiroug L, Saarma M, Kaila K, Rivera C. (2007) KCC2 interacts with the dendritic cytoskeleton to promote spine development. Neuron 56:1019-1033.
Martens JR, O’Connell K, Tamkun M. (2004) Targeting of ion channels to membrane microdomains: localization of KV channels to lipid rafts. Trends Pharmacol Sci 25:16-21.
Mercado A, Song L, Vazquez N, Mount DB, Gamba G. (2000) Functional comparison of the K+-Cl– cotransporters KCC1 and KCC4. J Biol Chem 275:30326–30334.
Mercado A, V?zquez N, Song L, Cort?s R, Enck AH, Welch R, Delpire E, Gamba G, Mount DB. (2005) NH2-terminal heterogeneity in the KCC3 K+-Cl- cotransporter. Am J Physiol Renal Physiol 289:F1246-F1261.
Mount DB, Mercado A, Song L, Xu J, George AL Jr, Delpire E, Gamba G. (1999) Cloning and characterization of KCC3 and KCC4, new members of the cation-chloride cotransporter gene family. J Biol Chem 274:16355-16362.
Overall CM, Kleifeld O. (2006) Tumour microenvironment - opinion: validating matrix metalloproteinases as drug targets and anti-targets for cancer therapy. Nat Rev Cancer 6:227-239.
Payne JA, Stevenson TJ, Donaldson LF. (1996) Molecular characterization of a putative K-Cl cotransporter in rat brain. A neuronal-specific isoform. J Biol Chem 271:16245-16252.
Payne JA. (1997) Functional characterization of the neuronal-specific K-Cl cotransporter: implications for [K+]o regulation. Am J Physiol 273:C1516-C1525.
Pearson MM, Lu J, Mount DB, Delpire E. (2001) Localization of the K+-Cl- cotransporter, KCC3, in the central and peripheral nervous systems: expression in the choroid plexus, large neurons and white matter tracts. Neuroscience 103:481-491.
Race JE, Makhlouf FN, Logue PJ, Wilson FH, Dunham PB, Holtzman EJ. (1999) Molecular cloning and functional characterization of KCC3, a new K-Cl cotransporter. Am J Physiol 277:C1210-C1219.
Renehan AG, Zwahlen M, Minder C, O'Dwyer ST, Shalet SM, Egger M. (2004) Insulin-like growth factor (IGF)-I, IGF binding protein-3, and cancer risk: systematic review and meta-regression analysis. Lancet 363:1346–1353.
Rinehart J, Maksimova YD, Tanis JE, Stone KL, Hodson CA, Zhang J, Risinger M, Pan W, Wu D, Colangelo CM, Forbush B, Joiner CH, Gulcicek EE, Gallagher PG, Lifton RP. (2009) Sites of regulated phosphorylation that control K-Cl cotransporter activity. Cell 138:525-536.
Ridley AJ, Schwartz MA, Burridge K, Firtel RA, Ginsberg MH, Borisy G, Parsons JT, Horwitz AR. (2003) Cell migration: integrating signals from front to back. Science 302(5651):1704-1709.
Ritter M. (1998) Cell volume regulatory ion transport in cell migration. Contrib Nephrol 123:135-157.
Rust MB, Faulhaber J, Budack MK, Pfeffer C, Maritzen T, Didi? M, Beck FX, Boettger T, Schubert R, Ehmke H, Jentsch TJ, H?bner CA. (2006) Neurogenic mechanisms contribute to hypertension in mice with disruption of the K-Cl cotransporter KCC3. Circ Res 98:549-556.
Rust MB, Alper SL, Rudhard Y, Shmukler BE, Vicente R, Brugnara C, Trudel M, Jentsch TJ, H?bner CA. (2007) Disruption of erythroid K-Cl cotransporters alters erythrocyte volume and partially rescues erythrocyte dehydration in SAD mice. J Clin Invest 117:1708-1717.
Sallinen R, Tornberg J, Putkiranta M, Horelli-Kuitunen N, Airaksinen MS, Wessman M. (2001) Chromosomal localization of SLC12A5/Slc12a5, the human and mouse genes for the neuron-specific K(+)-Cl(-) cotransporter (KCC2) defines a new region of conserved homology. Cytogenet Cell Genet 94:67-70.
Schoell WM, Janicek MF, Mirhashemi R. (1999) Epidemiology and biology of cervical cancer. Semin Surg Oncol 16:203-211.
Shen MR, Chou CY, Ellory JC. (2000) Volume-sensitive KCl cotransport associated with human cervical carcinogenesis. Pfluegers Arch 440:751-760.
Shen MR, Chou CY, Hsu KF, Liu HS, Dunham PB, Holtzman EJ, Ellory JC. (2001) The KCl cotransporter isoform KCC3 can play an important role in cell growth regulation. Proc Natl Acad Sci U S A 98:14714-14719.
Shen MR, Chou CY, Hsu KF, Hsu YM, Chiu WT, Tang MJ, Alper SL, Ellory JC. (2003) KCl cotransport is an important modulator of human cervical cancer growth and invasion. J Biol Chem 278:39941-39950.
Shen MR, Lin AC, Hsu YM, Chang TJ, Tang MJ, Alper SL, Ellory JC, Chou CY. (2004) Insulin-like growth factor 1 stimulates KCl cotransport, which is necessary for invasion and proliferation of cervical cancer and ovarian cancer cells. J Biol Chem 279:40017-40025.
Shen MR, Hsu YM, Hsu KF, Chen YF, Tang MJ, Chou CY. (2006) Insulin-like growth factor 1 is a potent stimulator of cervical cancer cell invasiveness and proliferation that is modulated by alphavbeta3 integrin signaling. Carcinogenesis 27:962-971.
Simons K, Ikonen E. (1997) Functional rafts in cell membranes. Nature 387:569-572.
Soldati T, Schliwa M. (2006) Powering membrane traffic in endocytosis and recycling. Nat Rev Mol Cell Biol 7:897-908.
Song L, Mercado A, V?zquez N, Xie Q, Desai R, George AL Jr, Gamba G, Mount DB. (2002) Molecular, functional, and genomic characterization of human KCC2, the neuronal K-Cl cotransporter. Brain Res Mol Brain Res 103:91-105.
Strange K, Singer TD, Morrison R, Delpire E. (2000) Dependence of KCC2 K-Cl cotransporter activity on a conserved carboxy terminus tyrosine residue. Am J Physiol 279:C860-C867.
Su W, Shmukler BE, Chernova MN, Stuart-Tilley AK, de Franceschi L, Brugnara C, Alper SL. (1999) Mouse K-Cl cotransporter KCC1: cloning, mapping, pathological expression, and functional regulation. Am J Physiol 277:C899-C912.
Syrjanen K, Kataja V, Yliskoski M, Chang F, Syrjanen S, Saarikoski S. (1992) Natural history of cervical human papillomavirus lesions does not substantiate the biologic relevance of the Bethesda System. Obstet Gynecol 79:675-682.
Thiery JP, Sleeman JP. (2006) Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol 7:131–142.
Toba G, Ohsako T, Miyata N, Ohtsuka T, Seong KH, Aigaki T. (1999) The gene search system: a method for efficient detection and rapid molecular identification of genes in Drosophila melanogaster. Genetics 151:725-737.
Velazquez H, Silva T. (2003) Cloning and localization of KCC4 in rabbit kidney: expression in distal convoluted tubule. Am J Physiol 285:F49-F58.
Wake H, Watanabe M, Moorhouse AJ, Kanematsu T, Horibe S, Matsukawa N, Asai K, Ojika K, Hirata M, Nabekura J. (2007) Early changes in KCC2 phosphorylation in response to neuronal stress result in functional downregulation. J Neurosci 27:1642-1650.
Webb DJ, Donais K, Whitmore LA, Thomas SM, Turner CE, Parsons JT, Horwitz AF. (2004) FAK-Src signalling through paxillin, ERK and MLCK regulates adhesion disassembly. Nat Cell Biol 6:154-161.
Wei WC, Lin HH, Shen MR, Tang MJ. (2008) Mechanosensing machinery for cells under low substratum rigidity. Am J Physiol Cell Physiol 295:C1579-1589.
Woo NS, Lu J, England R, McClellan R, Dufour S, Mount DB, Deutch AY, Lovinger DM, Delpire E. (2002) Hyperexcitability and epilepsy associated with disruption of the mouse neuronal-specific K-Cl cotransporter gene. Hippocampus 12:258-268.
Yamani MH, Tuzcu EM, Starling RC, Ratliff NB, Yu Y, Vince DG, Powell K, Cook D, McCarthy P, Young JB. (2002) Myocardial ischemic injury after heart transplantation is associated with upregulation of vitronectin receptor (?v?3), activation of the matrix metalloproteinase induction system, and subsequent development of coronary vasculopathy. Circulation 105:1955-1961.
Yeh HH, Lai WW, Chen HH, Liu HS, Su WC. Autocrine IL-6-induced Stat3 activation contributes to the pathogenesis of lung adenocarcinoma and malignant pleural effusion. Oncogene 2006; 25:4300-4309.
Yeo MG, Partridge MA, Ezratty EJ, Shen Q, Gundersen GG, Marcantonio EE. (2006) Src SH2 arginine 175 is required for cell motility: specific focal adhesion kinase targeting and focal adhesion assembly function. Mol Cell Biol 26:4399–4409.
Zamir E, Geiger B. (2001) Molecular complexity and dynamics of cell matrix adhesions. J Cell Sci 114:3583–90.
Zhang J, Lauf PK, Adragna NC. (2003) Platelet-derived growth factor regulates K-Cl cotransport in vascular smooth muscle cells. Am J Physiol 284:C674-C680.
zur Hausen H. (1991) Human papillomaviruses in the pathogenesis of anogenital cancer. Virology 184:9-13.