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標題: 14-3-3 sigma蛋白的胺基酸H180對調控Src活性及抑制肺癌進程的重要性
Amino acid H180 of 14-3-3sigma protein is important for regulating Src activity and inhibiting lung cancer progression
作者: 許峻豪
Hsu, Chun-Hao
關鍵字: lung cancer progression
14-3-3sigma
Src
14-3-3sigma
肺癌
Src
出版社: 生物醫學研究所
引用: Acloque, H., Adams, M. S., Fishwick, K., Bronner-Fraser, M., and Nieto, M. A. (2009). Epithelial-mesenchymal transitions: the importance of changing cell state in development and disease. The Journal of clinical investigation 119, 1438-1449. Agiostratidou, G., Hulit, J., Phillips, G. R., and Hazan, R. B. (2007). Differential cadherin expression: potential markers for epithelial to mesenchymal transformation during tumor progression. Journal of mammary gland biology and neoplasia 12, 127-133. Amato, P. A., Unanue, E. R., and Taylor, D. L. (1983). Distribution of actin in spreading macrophages: a comparative study on living and fixed cells. The Journal of cell biology 96, 750-761. Athwal, G. S., Lombardo, C. R., Huber, J. L., Masters, S. C., Fu, H., and Huber, S. C. (2000). Modulation of 14-3-3 protein interactions with target polypeptides by physical and metabolic effectors. Plant & cell physiology 41, 523-533. Babykutty, S., P, S. P., R, J. N., Kumar, M. A., Nair, M. S., Srinivas, P., and Gopala, S. (2011). Nimbolide retards tumor cell migration, invasion, and angiogenesis by downregulating MMP-2/9 expression via inhibiting ERK1/2 and reducing DNA-binding activity of NF-kappaB in colon cancer cells. Molecular carcinogenesis. Bach, P. B., Kattan, M. W., Thornquist, M. D., Kris, M. G., Tate, R. C., Barnett, M. J., Hsieh, L. J., and Begg, C. B. (2003). Variations in lung cancer risk among smokers. Journal of the National Cancer Institute 95, 470-478. Bartek, J., and Lukas, J. (2003). Chk1 and Chk2 kinases in checkpoint control and cancer. Cancer cell 3, 421-429. Birkenfeld, J., Betz, H., and Roth, D. (2003). Identification of cofilin and LIM-domain-containing protein kinase 1 as novel interaction partners of 14-3-3 zeta. The Biochemical journal 369, 45-54. Burns, S., Thrasher, A. J., Blundell, M. P., Machesky, L., and Jones, G. E. (2001). Configuration of human dendritic cell cytoskeleton by Rho GTPases, the WAS protein, and differentiation. Blood 98, 1142-1149. Chan, T. A., Hermeking, H., Lengauer, C., Kinzler, K. W., and Vogelstein, B. (1999). 14-3-3Sigma is required to prevent mitotic catastrophe after DNA damage. Nature 401, 616-620. Chen, W. T. (1989). Proteolytic activity of specialized surface protrusions formed at rosette contact sites of transformed cells. The Journal of experimental zoology 251, 167-185. Chen, W. T., Hasegawa, E., Hasegawa, T., Weinstock, C., and Yamada, K. M. (1985). Development of cell surface linkage complexes in cultured fibroblasts. The Journal of cell biology 100, 1103-1114. Cooley, M. E. (2000). Symptoms in adults with lung cancer. A systematic research review. Journal of pain and symptom management 19, 137-153. Cooley, M. E., Short, T. H., and Moriarty, H. J. (2003). Symptom prevalence, distress, and change over time in adults receiving treatment for lung cancer. Psycho-oncology 12, 694-708. Courtneidge, S. A., Azucena, E. F., Pass, I., Seals, D. F., and Tesfay, L. (2005). The SRC substrate Tks5, podosomes (invadopodia), and cancer cell invasion. Cold Spring Harbor symposia on quantitative biology 70, 167-171. Datta, S. R., Katsov, A., Hu, L., Petros, A., Fesik, S. W., Yaffe, M. B., and Greenberg, M. E. (2000). 14-3-3 proteins and survival kinases cooperate to inactivate BAD by BH3 domain phosphorylation. Molecular cell 6, 41-51. David-Pfeuty, T., and Singer, S. J. (1980). Altered distributions of the cytoskeletal proteins vinculin and alpha-actinin in cultured fibroblasts transformed by Rous sarcoma virus. Proceedings of the National Academy of Sciences of the United States of America 77, 6687-6691. Du, C., Zhang, C., Hassan, S., Biswas, M. H., and Balaji, K. C. (2010). Protein kinase D1 suppresses epithelial-to-mesenchymal transition through phosphorylation of snail. Cancer research 70, 7810-7819. Dudek, S. M., and Garcia, J. G. (2001). Cytoskeletal regulation of pulmonary vascular permeability. J Appl Physiol 91, 1487-1500. Eckert, M. A., and Yang, J. (2011). Targeting invadopodia to block breast cancer metastasis. Oncotarget 2, 562-568. Fisseler-Eckhoff, A. (2009). [New TNM classification of malignant lung tumors 2009 from a pathology perspective]. Der Pathologe 30 Suppl 2, 193-199. Frame, M. C. (2002). Src in cancer: deregulation and consequences for cell behaviour. Biochimica et biophysica acta 1602, 114-130. Fu, H., Subramanian, R. R., and Masters, S. C. (2000). 14-3-3 proteins: structure, function, and regulation. Annual review of pharmacology and toxicology 40, 617-647. Fukuoka, M., Yano, S., Giaccone, G., Tamura, T., Nakagawa, K., Douillard, J. Y., Nishiwaki, Y., Vansteenkiste, J., Kudoh, S., Rischin, D., et al. (2003). Multi-institutional randomized phase II trial of gefitinib for previously treated patients with advanced non-small-cell lung cancer (The IDEAL 1 Trial) [corrected]. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 21, 2237-2246. Gao, Y., Jiang, M., Yang, T., Ni, J., and Chen, J. (2006). A Cdc2-related protein kinase hPFTAIRE1 from human brain interacting with 14-3-3 proteins. Cell research 16, 539-547. Gay, B., Furet, P., Garcia-Echeverria, C., Rahuel, J., Chene, P., Fretz, H., Schoepfer, J., and Caravatti, G. (1997). Dual specificity of Src homology 2 domains for phosphotyrosine peptide ligands. Biochemistry 36, 5712-5718. Geiger, T. R., and Peeper, D. S. (2009). Metastasis mechanisms. Biochimica et biophysica acta 1796, 293-308. Giannopoulou, E., Dimitropoulos, K., Argyriou, A. A., Koutras, A. K., Dimitrakopoulos, F., and Kalofonos, H. P. (2010). An in vitro study, evaluating the effect of sunitinib and/or lapatinib on two glioma cell lines. Investigational new drugs 28, 554-560. Gift, A. G., Jablonski, A., Stommel, M., and Given, C. W. (2004). Symptom clusters in elderly patients with lung cancer. Oncology nursing forum 31, 202-212. Gimona, M., Buccione, R., Courtneidge, S. A., and Linder, S. (2008). Assembly and biological role of podosomes and invadopodia. Current opinion in cell biology 20, 235-241. Guarino, M. (2010). Src signaling in cancer invasion. Journal of cellular physiology 223, 14-26. Helton, E. S., and Chen, X. (2007). p53 modulation of the DNA damage response. Journal of cellular biochemistry 100, 883-896. Herner, A., Sauliunaite, D., Michalski, C. W., Erkan, M., De Oliveira, T., Abiatari, I., Kong, B., Esposito, I., Friess, H., and Kleeff, J. (2011). Glutamate increases pancreatic cancer cell invasion and migration via AMPA receptor activation and Kras-MAPK signaling. International journal of cancer Journal international du cancer 129, 2349-2359. Hoffman, P. C., Mauer, A. M., and Vokes, E. E. (2000). Lung cancer. Lancet 355, 479-485. Hotulainen, P., and Lappalainen, P. (2006). Stress fibers are generated by two distinct actin assembly mechanisms in motile cells. The Journal of cell biology 173, 383-394. Huveneers, S., and Danen, E. H. (2009). Adhesion signaling - crosstalk between integrins, Src and Rho. Journal of cell science 122, 1059-1069. Iwata, N., Yamamoto, H., Sasaki, S., Itoh, F., Suzuki, H., Kikuchi, T., Kaneto, H., Iku, S., Ozeki, I., Karino, Y., et al. (2000). Frequent hypermethylation of CpG islands and loss of expression of the 14-3-3 sigma gene in human hepatocellular carcinoma. Oncogene 19, 5298-5302. Jemal, A., Thomas, A., Murray, T., and Thun, M. (2002). Cancer statistics, 2002. CA: a cancer journal for clinicians 52, 23-47. Jeremic, B., Shibamoto, Y., Acimovic, L., and Milisavljevic, S. (1996). Hyperfractionated radiation therapy with or without concurrent low-dose daily carboplatin/etoposide for stage III non-small-cell lung cancer: a randomized study. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 14, 1065-1070. Jin, Y. H., Kim, Y. J., Kim, D. W., Baek, K. H., Kang, B. Y., Yeo, C. Y., and Lee, K. Y. (2008). Sirt2 interacts with 14-3-3 beta/gamma and down-regulates the activity of p53. Biochemical and biophysical research communications 368, 690-695. Kalluri, R., and Weinberg, R. A. (2009). The basics of epithelial-mesenchymal transition. The Journal of clinical investigation 119, 1420-1428. Keller, S. M., Adak, S., Wagner, H., Herskovic, A., Komaki, R., Brooks, B. J., Perry, M. C., Livingston, R. B., and Johnson, D. H. (2000). A randomized trial of postoperative adjuvant therapy in patients with completely resected stage II or IIIA non-small-cell lung cancer. Eastern Cooperative Oncology Group. The New England journal of medicine 343, 1217-1222. Kelley, L. C., Ammer, A. G., Hayes, K. E., Martin, K. H., Machida, K., Jia, L., Mayer, B. J., and Weed, S. A. (2010). Oncogenic Src requires a wild-type counterpart to regulate invadopodia maturation. Journal of cell science 123, 3923-3932. Ko, B. S., Chang, T. C., Hsu, C., Chen, Y. C., Shen, T. L., Chen, S. C., Wang, J., Wu, K. K., Jan, Y. J., and Liou, J. Y. (2011). Overexpression of 14-3-3epsilon predicts tumour metastasis and poor survival in hepatocellular carcinoma. Histopathology 58, 705-711. Koch, C. A., Anderson, D., Moran, M. F., Ellis, C., and Pawson, T. (1991). SH2 and SH3 domains: elements that control interactions of cytoplasmic signaling proteins. Science 252, 668-674. Kruse, J. P., and Gu, W. (2009). Modes of p53 regulation. Cell 137, 609-622. Kuo, L., Chang, H. C., Leu, T. H., Maa, M. C., and Hung, W. C. (2006). Src oncogene activates MMP-2 expression via the ERK/Sp1 pathway. Journal of cellular physiology 207, 729-734. Kurup, A., and Hanna, N. H. (2004). Treatment of small cell lung cancer. Critical reviews in oncology/hematology 52, 117-126. Lafleur, M. A., Handsley, M. M., and Edwards, D. R. (2003). Metalloproteinases and their inhibitors in angiogenesis. Expert reviews in molecular medicine 5, 1-39. Lai, F. P., Szczodrak, M., Block, J., Faix, J., Breitsprecher, D., Mannherz, H. G., Stradal, T. E., Dunn, G. A., Small, J. V., and Rottner, K. (2008). Arp2/3 complex interactions and actin network turnover in lamellipodia. The EMBO journal 27, 982-992. Lambrechts, A., Van Troys, M., and Ampe, C. (2004). The actin cytoskeleton in normal and pathological cell motility. The international journal of biochemistry & cell biology 36, 1890-1909. Laronga, C., Yang, H. Y., Neal, C., and Lee, M. H. (2000). Association of the cyclin-dependent kinases and 14-3-3 sigma negatively regulates cell cycle progression. The Journal of biological chemistry 275, 23106-23112. Lee, J. S., and Gotlieb, A. I. (2003). Understanding the role of the cytoskeleton in the complex regulation of the endothelial repair. Histology and histopathology 18, 879-887. Lee, T. Y., and Gotlieb, A. I. (2002). Rho and basic fibroblast growth factor involvement in centrosome redistribution and actin microfilament remodeling during early endothelial wound repair. Journal of vascular surgery : official publication, the Society for Vascular Surgery [and] International Society for Cardiovascular Surgery, North American Chapter 35, 1242-1252. Leffers, H., Madsen, P., Rasmussen, H. H., Honore, B., Andersen, A. H., Walbum, E., Vandekerckhove, J., and Celis, J. E. (1993). Molecular cloning and expression of the transformation sensitive epithelial marker stratifin. A member of a protein family that has been involved in the protein kinase C signalling pathway. Journal of molecular biology 231, 982-998. Li, D. J., Deng, G., Xiao, Z. Q., Yao, H. X., Li, C., Peng, F., Li, M. Y., Zhang, P. F., Chen, Y. H., and Chen, Z. C. (2009a). Identificating 14-3-3 sigma as a lymph node metastasis-related protein in human lung squamous carcinoma. Cancer letters 279, 65-73. Li, Z., Liu, J. Y., and Zhang, J. T. (2009b). 14-3-3sigma, the double-edged sword of human cancers. American journal of translational research 1, 326-340. Li, Z., Zhao, J., Du, Y., Park, H. R., Sun, S. Y., Bernal-Mizrachi, L., Aitken, A., Khuri, F. R., and Fu, H. (2008). Down-regulation of 14-3-3zeta suppresses anchorage-independent growth of lung cancer cells through anoikis activation. Proceedings of the National Academy of Sciences of the United States of America 105, 162-167. Light, Y., Paterson, H., and Marais, R. (2002). 14-3-3 antagonizes Ras-mediated Raf-1 recruitment to the plasma membrane to maintain signaling fidelity. Molecular and cellular biology 22, 4984-4996. Linder, S. (2007). The matrix corroded: podosomes and invadopodia in extracellular matrix degradation. Trends in cell biology 17, 107-117. Liu, H., Ong, S. E., Badu-Nkansah, K., Schindler, J., White, F. M., and Hynes, R. O. (2011). CUB-domain-containing protein 1 (CDCP1) activates Src to promote melanoma metastasis. Proceedings of the National Academy of Sciences of the United States of America 108, 1379-1384. Liu, Y. N., Lee, W. W., Wang, C. Y., Chao, T. H., Chen, Y., and Chen, J. H. (2005). Regulatory mechanisms controlling human E-cadherin gene expression. Oncogene 24, 8277-8290. Lu, J., Guo, H., Treekitkarnmongkol, W., Li, P., Zhang, J., Shi, B., Ling, C., Zhou, X., Chen, T., Chiao, P. J., et al. (2009). 14-3-3zeta Cooperates with ErbB2 to promote ductal carcinoma in situ progression to invasive breast cancer by inducing epithelial-mesenchymal transition. Cancer cell 16, 195-207. Luo, Y., Xu, X., Lele, T., Kumar, S., and Ingber, D. E. (2008). A multi-modular tensegrity model of an actin stress fiber. Journal of biomechanics 41, 2379-2387. Luxenburg, C., Parsons, J. T., Addadi, L., and Geiger, B. (2006). Involvement of the Src-cortactin pathway in podosome formation and turnover during polarization of cultured osteoclasts. Journal of cell science 119, 4878-4888. Massague, J. (2008). TGFbeta in Cancer. Cell 134, 215-230. Mattila, P. K., and Lappalainen, P. (2008). Filopodia: molecular architecture and cellular functions. Nature reviews Molecular cell biology 9, 446-454. Mikami, S., Katsube, K., Oya, M., Ishida, M., Kosaka, T., Mizuno, R., Mukai, M., and Okada, Y. (2011). Expression of Snail and Slug in renal cell carcinoma: E-cadherin repressor Snail is associated with cancer invasion and prognosis. Laboratory investigation; a journal of technical methods and pathology 91, 1443-1458. Mongiu, A. K., Weitzke, E. L., Chaga, O. Y., and Borisy, G. G. (2007). Kinetic-structural analysis of neuronal growth cone veil motility. Journal of cell science 120, 1113-1125. Moreira, J. M., Gromov, P., and Celis, J. E. (2004). Expression of the tumor suppressor protein 14-3-3 sigma is down-regulated in invasive transitional cell carcinomas of the urinary bladder undergoing epithelial-to-mesenchymal transition. Molecular & cellular proteomics : MCP 3, 410-419. Murphy, D. A., and Courtneidge, S. A. (2011). The ''ins'' and ''outs'' of podosomes and invadopodia: characteristics, formation and function. Nature reviews Molecular cell biology 12, 413-426. Ng, Y. H., Zhu, H., and Leung, P. C. (2011). Twist Modulates Human Trophoblastic Cell Invasion via Regulation of N-Cadherin. Endocrinology. Onder, T. T., Gupta, P. B., Mani, S. A., Yang, J., Lander, E. S., and Weinberg, R. A. (2008). Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer research 68, 3645-3654. Patel, J. C., Hall, A., and Caron, E. (2002). Vav regulates activation of Rac but not Cdc42 during FcgammaR-mediated phagocytosis. Molecular biology of the cell 13, 1215-1226. Raftopoulou, M., and Hall, A. (2004). Cell migration: Rho GTPases lead the way. Developmental biology 265, 23-32. Saltel, F., Destaing, O., Bard, F., Eichert, D., and Jurdic, P. (2004). Apatite-mediated actin dynamics in resorbing osteoclasts. Molecular biology of the cell 15, 5231-5241. Seals, D. F., Azucena, E. F., Jr., Pass, I., Tesfay, L., Gordon, R., Woodrow, M., Resau, J. H., and Courtneidge, S. A. (2005). The adaptor protein Tks5/Fish is required for podosome formation and function, and for the protease-driven invasion of cancer cells. Cancer cell 7, 155-165. Sengupta, S., and Harris, C. C. (2005). p53: traffic cop at the crossroads of DNA repair and recombination. Nature reviews Molecular cell biology 6, 44-55. Shook, D., and Keller, R. (2003). Mechanisms, mechanics and function of epithelial-mesenchymal transitions in early development. Mechanisms of development 120, 1351-1383. Siegel, R., Naishadham, D., and Jemal, A. (2012). Cancer statistics, 2012. CA: a cancer journal for clinicians 62, 10-29. Songyang, Z., Shoelson, S. E., Chaudhuri, M., Gish, G., Pawson, T., Haser, W. G., King, F., Roberts, T., Ratnofsky, S., Lechleider, R. J., and et al. (1993). SH2 domains recognize specific phosphopeptide sequences. Cell 72, 767-778. Steinmetz, M. O., Goldie, K. N., and Aebi, U. (1997). A correlative analysis of actin filament assembly, structure, and dynamics. The Journal of cell biology 138, 559-574. Stradal, T. E., Rottner, K., Disanza, A., Confalonieri, S., Innocenti, M., and Scita, G. (2004). Regulation of actin dynamics by WASP and WAVE family proteins. Trends in cell biology 14, 303-311. Sudol, M. (2011). From Rous sarcoma virus to plasminogen activator, src oncogene and cancer management. Oncogene 30, 3003-3010. Sun, G., Ramdas, L., Wang, W., Vinci, J., McMurray, J., and Budde, R. J. (2002). Effect of autophosphorylation on the catalytic and regulatory properties of protein tyrosine kinase Src. Archives of biochemistry and biophysics 397, 11-17. Sun, G., Sharma, A. K., and Budde, R. J. (1998). Autophosphorylation of Src and Yes blocks their inactivation by Csk phosphorylation. Oncogene 17, 1587-1595. Suzuki, H., Itoh, F., Toyota, M., Kikuchi, T., Kakiuchi, H., and Imai, K. (2000). Inactivation of the 14-3-3 sigma gene is associated with 5'' CpG island hypermethylation in human cancers. Cancer research 60, 4353-4357. Takihara, Y., Matsuda, Y., and Hara, J. (2000). Role of the beta isoform of 14-3-3 proteins in cellular proliferation and oncogenic transformation. Carcinogenesis 21, 2073-2077. Tanoue, L. T., and Detterbeck, F. C. (2009). New TNM classification for non-small-cell lung cancer. Expert review of anticancer therapy 9, 413-423. Tarone, G., Cirillo, D., Giancotti, F. G., Comoglio, P. M., and Marchisio, P. C. (1985). Rous sarcoma virus-transformed fibroblasts adhere primarily at discrete protrusions of the ventral membrane called podosomes. Experimental cell research 159, 141-157. Tatin, F., Varon, C., Genot, E., and Moreau, V. (2006). A signalling cascade involving PKC, Src and Cdc42 regulates podosome assembly in cultured endothelial cells in response to phorbol ester. Journal of cell science 119, 769-781. Thiery, J. P. (2003). Epithelial-mesenchymal transitions in development and pathologies. Current opinion in cell biology 15, 740-746. Tokuo, H., and Ikebe, M. (2004). Myosin X transports Mena/VASP to the tip of filopodia. Biochemical and biophysical research communications 319, 214-220. Tsuchiya, E., Nakamura, Y., Weng, S. Y., Nakagawa, K., Tsuchiya, S., Sugano, H., and Kitagawa, T. (1992). Allelotype of non-small cell lung carcinoma--comparison between loss of heterozygosity in squamous cell carcinoma and adenocarcinoma. Cancer research 52, 2478-2481. Umbricht, C. B., Evron, E., Gabrielson, E., Ferguson, A., Marks, J., and Sukumar, S. (2001). Hypermethylation of 14-3-3 sigma (stratifin) is an early event in breast cancer. Oncogene 20, 3348-3353. Van Der Hoeven, P. C., Van Der Wal, J. C., Ruurs, P., and Van Blitterswijk, W. J. (2000). Protein kinase C activation by acidic proteins including 14-3-3. The Biochemical journal 347 Pt 3, 781-785. van Hemert, M. J., Steensma, H. Y., and van Heusden, G. P. (2001). 14-3-3 proteins: key regulators of cell division, signalling and apoptosis. BioEssays : news and reviews in molecular, cellular and developmental biology 23, 936-946. Wang, Y. (2010). Breast cancer metastasis driven by ErbB2 and 14-3-3zeta: A division of labor. Cell adhesion & migration 4, 7-9. Wheeler, D. L., Iida, M., and Dunn, E. F. (2009). The role of Src in solid tumors. The oncologist 14, 667-678. Wiercinska, E., Naber, H. P., Pardali, E., van der Pluijm, G., van Dam, H., and ten Dijke, P. (2011). The TGF-beta/Smad pathway induces breast cancer cell invasion through the up-regulation of matrix metalloproteinase 2 and 9 in a spheroid invasion model system. Breast cancer research and treatment 128, 657-666. Wilker, E. W., Grant, R. A., Artim, S. C., and Yaffe, M. B. (2005). A structural basis for 14-3-3sigma functional specificity. The Journal of biological chemistry 280, 18891-18898. Woodhouse, E. C., Chuaqui, R. F., and Liotta, L. A. (1997). General mechanisms of metastasis. Cancer 80, 1529-1537. Yaffe, M. B., Rittinger, K., Volinia, S., Caron, P. R., Aitken, A., Leffers, H., Gamblin, S. J., Smerdon, S. J., and Cantley, L. C. (1997). The structural basis for 14-3-3:phosphopeptide binding specificity. Cell 91, 961-971. Yamazaki, D., Kurisu, S., and Takenawa, T. (2005). Regulation of cancer cell motility through actin reorganization. Cancer science 96, 379-386. Yang, J., and Weinberg, R. A. (2008). Epithelial-mesenchymal transition: at the crossroads of development and tumor metastasis. Developmental cell 14, 818-829. Yokotsuka, M., Iwaya, K., Saito, T., Pandiella, A., Tsuboi, R., Kohno, N., Matsubara, O., and Mukai, K. (2011). Overexpression of HER2 signaling to WAVE2-Arp2/3 complex activates MMP-independent migration in breast cancer. Breast cancer research and treatment 126, 311-318. Zeisberg, M., and Neilson, E. G. (2009). Biomarkers for epithelial-mesenchymal transitions. The Journal of clinical investigation 119, 1429-1437. Zhang, X. H., Wang, Q., Gerald, W., Hudis, C. A., Norton, L., Smid, M., Foekens, J. A., and Massague, J. (2009). Latent bone metastasis in breast cancer tied to Src-dependent survival signals. Cancer cell 16, 67-78. Zhang, Y. E. (2009). Non-Smad pathways in TGF-beta signaling. Cell research 19, 128-139.
摘要: The family of 14-3-3 proteins is implicated with many biological activities by binding to other proteins. It consists of seven known highly conserved isoforms (β, γ, ε, ζ, η, σ, and τ) and most of them are identified as oncogenic functions except for 14-3-3sigma, a well-known tumor suppressor. To investigate the suppressor characteristic of 14-3-3sigma divergent from others, we analyzed the protein sequences of 14-3-3 family and found that the major difference between 14-3-3sigma and its oncogenic members was due to loss of the phospho-site at His180. Thus, H180Y mutation was generated by site-direct mutagenesis to determine the role of this amino acid in the suppressive characteristic of 14-3-3sigma. First, we showed that overexpression of 14-3-3sigma could decrease the abilities of cancer cell invasion, migration and proliferation, as well as repress cell scattering. On the contrary, H180Y not only induced cell surface protrusions and branchings but also switched 14-3-3sigma from suppressor to oncogene. Meanwhile, H180Y mutant also generates a SH2-binding motif (YYEI) for Src binding. The results of co-immunoprecipitation demonstrated that the association between H180Y mutant and Src was higher than that of wild type, indicating the importance of H180 site for preventing 14-3-3sigma from binding to Src. We also observed that overexpression of H180Y mutant enhanced cell invasion capability and MMP2 secretion. induced the formation of podosome rings and led to matrix degradation. Finally, we demonstrated that H180Y overexpression enhanced cell migration, colony formation, proliferation in vitro, and tumorigenesis in vivo. Taken together, our findings suggest that amino acid H180 of 14-3-3sigma is an essential site to disrupt the 14-3-3sigma-Src interaction and plays important roles in the regulation of Src activity and the tumor suppression of 14-3-3sigma.
14-3-3家族成員利用與相關蛋白質進行交互作用來調控生物活性。在哺乳動物當中,14-3-3家族被發現有七個家族成員,分別為β、γ、ε、ζ、 η、σ和τ,而這些家族成員大部分被視為致癌基因,除了14-3-3sigma (YWHAS)被定義為是一個腫瘤抑制基因。為了瞭解14-3-3sigma與其他家族成員的差異,我們進行所有家族成員的蛋白質序列分析,發現14-3-3sigma與其他致癌基因的家族成員最主要的差異是在第180個胺基酸,由可被磷酸化的酪胺酸取代為組胺酸。因此,我們利用單點突變方法構築H180Y突變株,以探討在14-3-3sigma腫瘤抑制特性中此胺基酸的角色。研究發現,大量表現14-3-3sigma會抑制細胞的移動、侵襲與生長能力。然而,突變的H180Y蛋白質不只會增加細胞的突出及分支情形,也會將原有的腫瘤抑制基因功能轉變成致癌性。此外,突變的H180Y會獲得與其他家族成員同樣擁有的SH2結合區域(YYEI)。我們利用免疫沉澱實驗發現,突變的H180Y與野生性的YWHAS相比,的確會提升與Src之間的交互作用,顯示H180的位置對於14-3-3sigma與Src結合是重要的。我們觀察到大量表現帶有H180Y的構築,會促進細胞的侵犯能力和導致MMP2的分泌;突變的H180Y也會誘導podosome環的形成,造成瓦解細胞基質能力的提升。最後,研究也證實大量表現突變的H180Y會促進細胞的移動、聚落形成、生長以及腫瘤形成。總而言之,由我們的結果推測14-3-3sigma的第180個胺基酸對於其與Src的交互作用是必要的,且在其調控Src活性及抑制肺癌進程中扮演重要的角色。
URI: http://hdl.handle.net/11455/20166
其他識別: U0005-0702201200250000
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-0702201200250000
顯示於類別:生物醫學研究所

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