臺灣博碩士論文加值系統

中文摘要
Troglitazone (簡稱TGZ)是一個PPARg誘發劑而且具有抗癌的潛力(包括抑制癌細胞生長和誘發其凋亡)。但是對於TGZ影響癌細胞移動的研究並不多。因此本篇論文主旨在於闡明TGZ對卵巢癌細胞，ES-2，移動能力的影響。我們以wound healing assay和transwell chamber assay證明TGZ可以隨濃度和時間增加而抑制細胞移動。TGZ會影響幾個細胞移動所必需的步驟，包括：細胞伸展、附著點的形成和FAK的自我磷酸化。雖然TGZ可以促進PPAR的表現，但是以PPARg siRNA抑制PPARg表現後，並不影響TGZ抑制細胞移動的能力。此外，TGZ可以隨濃度和時間增加而抑制FAK的自我磷酸化，符合TGZ抑制細胞移動的現象。TGZ亦隨濃度和時間增加而抑制PTEN C端的磷酸化；細胞在處理TGZ (濃度大於10uM)後，可以使PTEN的分布集中在細胞膜上，這些結果顯示TGZ可以活化PTEN，於是推測TGZ可以經由活化PTEN而抑制FAK的自我磷酸化和細胞移動。本篇論文結果顯示，TGZ可以抑制卵巢癌細胞移動，可能和TGZ調控FAK和PTEN的活性有關，未來也許能進一步應用在治療卵巢癌上。

Abstract
Troglitazone (TGZ) is a peroxisome proliferator-activated receptor gamma (PPARg) agonist and a potential anticancer agent (inducer of mitotic arrest and apoptosis in cancer cells). Little is known about the effect of this agent on cancer cell migration. The present study aims to clarify the effect of TGZ on cell motility in a human ovarian carcinoma cell line, ES-2. We demonstrated that TGZ dose- and time-dependently impairs cell migration as assessed by wound-healing and Boyden chamber transwell experiments. TGZ treatment impaired several processes essential for cell migration (including cell spreading, stress fiber formation, tyrosine phosphorylation of focal adhesion kinase [FAK], and focal adhesion assembly in cells grown on fibronectin substratum). TGZ induced PPARg overexpression in ES-2 cells, but abolishing PPARg expression by small interfering RNA had no influence on TGZ-mediated anti-migratory effects. In addition, TGZ dose- and time-dependently suppressed FAK autophosphorylation, which correlated with the degree of TGZ inhibition of cell migration. TGZ also dose- and time-dependently suppressed phosphorylation of the C-terminal of PTEN (phosphatase and tensin homologue deleted on chromosome ten), an identified migration regulator. Cells treated with TGZ (higher than 10 uM) caused accumulation of PTEN in plasma membrane, a sign of PTEN activation, suggesting the involvement of PTEN in TGZ inhibition of FAK phosphorylation and cell migration. In sum, our data suggest a mechanism for TZG’s inhibition of cell migration, involving the regulation of FAK and PTEN activity, and a possible treatment for human ovarian cancer based on the anti-migration potential of PPARγ agonists.

目錄
目錄
表目錄
圖目錄
縮寫表
中文摘要
英文摘要
第一章、 緒論
第二章、 實驗材料和方法
第三章、 結果
第四章、 討論
第五章、 參考文獻
第六章、 圖

Aronsohn, M. S., H. M. Brown, et al. (2003). "Expression of focal adhesion kinase and phosphorylated focal adhesion kinase in squamous cell carcinoma of the larynx." Laryngoscope 113(11): 1944-8.
Cance, W. G., J. E. Harris, et al. (2000). "Immunohistochemical analyses of focal adhesion kinase expression in benign and malignant human breast and colon tissues: correlation with preinvasive and invasive phenotypes." Clin Cancer Res 6(6): 2417-23.
Cary, L. A., J. F. Chang, et al. (1996). "Stimulation of cell migration by overexpression of focal adhesion kinase and its association with Src and Fyn." J Cell Sci 109 ( Pt 7): 1787-94.
Casey, R. C. and A. P. Skubitz (2000). "CD44 and beta1 integrins mediate ovarian carcinoma cell migration toward extracellular matrix proteins." Clin Exp Metastasis 18(1): 67-75.
Chen, Y., S. M. Wang, et al. (2006). "Effects of PPARgamma agonists on cell survival and focal adhesions in a Chinese thyroid carcinoma cell line." J Cell Biochem 98(4): 1021-35.
Cheney, I. W., D. E. Johnson, et al. (1998). "Suppression of tumorigenicity of glioblastoma cells by adenovirus-mediated MMAC1/PTEN gene transfer." Cancer Res 58(11): 2331-4.
Cuevas, B. D., A. N. Abell, et al. (2003). "MEKK1 regulates calpain-dependent proteolysis of focal adhesion proteins for rear-end detachment of migrating fibroblasts." Embo J 22(13): 3346-55.
Das, S., J. E. Dixon, et al. (2003). "Membrane-binding and activation mechanism of PTEN." Proc Natl Acad Sci U S A 100(13): 7491-6.
Furnari, F. B., H. Lin, et al. (1997). "Growth suppression of glioma cells by PTEN requires a functional phosphatase catalytic domain." Proc Natl Acad Sci U S A 94(23): 12479-84.
Galli, A., E. Ceni, et al. (2004). "Antidiabetic thiazolidinediones inhibit invasiveness of pancreatic cancer cells via PPARgamma independent mechanisms." Gut 53(11): 1688-97.
Grisaru-Granovsky, S., Z. Salah, et al. (2005). "Differential expression of protease activated receptor 1 (Par1) and pY397FAK in benign and malignant human ovarian tissue samples." Int J Cancer 113(3): 372-8.
Guo, W. and F. G. Giancotti (2004). "Integrin signalling during tumour progression." Nat Rev Mol Cell Biol 5(10): 816-26.
Hamadi, A., M. Bouali, et al. (2005). "Regulation of focal adhesion dynamics and disassembly by phosphorylation of FAK at tyrosine 397." J Cell Sci 118(Pt 19): 4415-25.
Hattori, Y., S. Hattori, et al. (1999). "Troglitazone upregulates nitric oxide synthesis in vascular smooth muscle cells." Hypertension 33(4): 943-8.
Hlobilkova, A., J. Knillova, et al. (2003). "The mechanism of action of the tumour suppressor gene PTEN." Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 147(1): 19-25.
Hsia, D. A., S. K. Mitra, et al. (2003). "Differential regulation of cell motility and invasion by FAK." J Cell Biol 160(5): 753-67.
Huang, M. E., Y. C. Ye, et al. (1988). "Use of all-trans retinoic acid in the treatment of acute promyelocytic leukemia." Blood 72(2): 567-72.
Ilic, D., Y. Furuta, et al. (1995). "Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-deficient mice." Nature 377(6549): 539-44.
Ilic, D., O. Genbacev, et al. (2001). "Plasma membrane-associated pY397FAK is a marker of cytotrophoblast invasion in vivo and in vitro." Am J Pathol 159(1): 93-108.
Jemal, A., R. C. Tiwari, et al. (2004). "Cancer statistics, 2004." CA Cancer J Clin 54(1): 8-29.
Judson, P. L., X. He, et al. (1999). "Overexpression of focal adhesion kinase, a protein tyrosine kinase, in ovarian carcinoma." Cancer 86(8): 1551-6.
Kersten, S., B. Desvergne, et al. (2000). "Roles of PPARs in health and disease." Nature 405(6785): 421-4.
Kohn, E. C., L. A. Travers, et al. (2005). "Malignant effusions are sources of fibronectin and other promigratory and proinvasive components." Diagn Cytopathol 33(5): 300-8.
Kokenyesi, R., K. P. Murray, et al. (2003). "Invasion of interstitial matrix by a novel cell line from primary peritoneal carcinosarcoma, and by established ovarian carcinoma cell lines: role of cell-matrix adhesion molecules, proteinases, and E-cadherin expression." Gynecol Oncol 89(1): 60-72.
Kornberg, L. J. (1998). "Focal adhesion kinase and its potential involvement in tumor invasion and metastasis." Head Neck 20(8): 745-52.
Lee, K. S., J. H. Park, et al. (2006). "Troglitazone inhibits endothelial cell proliferation through suppression of casein kinase 2 activity." Biochem Biophys Res Commun 346(1): 83-8.
Li, J., C. Yen, et al. (1997). "PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostate cancer." Science 275(5308): 1943-7.
Li, Z., X. Dong, et al. (2005). "Regulation of PTEN by Rho small GTPases." Nat Cell Biol 7(4): 399-404.
Liu, J., H. Lu, et al. (2005). "Peroxisome proliferator activated receptor-gamma ligands induced cell growth inhibition and its influence on matrix metalloproteinase activity in human myeloid leukemia cells." Cancer Chemother Pharmacol 56(4): 400-8.
Masuda, T., K. Wada, et al. (2005). "Critical role of peroxisome proliferator-activated receptor gamma on anoikis and invasion of squamous cell carcinoma." Clin Cancer Res 11(11): 4012-21.
McLean, G. W., N. O. Carragher, et al. (2005). "The role of focal-adhesion kinase in cancer - a new therapeutic opportunity." Nat Rev Cancer 5(7): 505-15.
Mitra, S. K., D. A. Hanson, et al. (2005). "Focal adhesion kinase: in command and control of cell motility." Nat Rev Mol Cell Biol 6(1): 56-68.
Miyazaki, T., H. Kato, et al. (2003). "FAK overexpression is correlated with tumour invasiveness and lymph node metastasis in oesophageal squamous cell carcinoma." Br J Cancer 89(1): 140-5.
Motomura, W., M. Nagamine, et al. (2004). "Inhibition of cell invasion and morphological change by troglitazone in human pancreatic cancer cells." J Gastroenterol 39(5): 461-8.
Owens, L. V., L. Xu, et al. (1995). "Overexpression of the focal adhesion kinase (p125FAK) in invasive human tumors." Cancer Res 55(13): 2752-5.
Owens, L. V., L. Xu, et al. (1996). "Focal adhesion kinase as a marker of invasive potential in differentiated human thyroid cancer." Ann Surg Oncol 3(1): 100-5.
Parsons, J. T., K. H. Martin, et al. (2000). "Focal adhesion kinase: a regulator of focal adhesion dynamics and cell movement." Oncogene 19(49): 5606-13.
Patel, L., I. Pass, et al. (2001). "Tumor suppressor and anti-inflammatory actions of PPARgamma agonists are mediated via upregulation of PTEN." Curr Biol 11(10): 764-8.
Rieppi, M., V. Vergani, et al. (1999). "Mesothelial cells induce the motility of human ovarian carcinoma cells." Int J Cancer 80(2): 303-7.
Rigacci, S., E. Rovida, et al. (2002). "Low Mr phosphotyrosine protein phosphatase associates and dephosphorylates p125 focal adhesion kinase, interfering with cell motility and spreading." J Biol Chem 277(44): 41631-6.
Rosen, E. D., P. Sarraf, et al. (1999). "PPAR gamma is required for the differentiation of adipose tissue in vivo and in vitro." Mol Cell 4(4): 611-7.
Rovin, J. D., H. F. Frierson, Jr., et al. (2002). "Expression of focal adhesion kinase in normal and pathologic human prostate tissues." Prostate 53(2): 124-32.
Sawai, H., J. Liu, et al. (2006). "Activation of peroxisome proliferator-activated receptor-gamma decreases pancreatic cancer cell invasion through modulation of the plasminogen activator system." Mol Cancer Res 4(3): 159-67.
Schaefer, K. L., K. Wada, et al. (2005). "Peroxisome proliferator-activated receptor gamma inhibition prevents adhesion to the extracellular matrix and induces anoikis in hepatocellular carcinoma cells." Cancer Res 65(6): 2251-9.
Schaller, M. D. (2001). "Biochemical signals and biological responses elicited by the focal adhesion kinase." Biochim Biophys Acta 1540(1): 1-21.
Schaller, M. D., J. D. Hildebrand, et al. (1994). "Autophosphorylation of the focal adhesion kinase, pp125FAK, directs SH2-dependent binding of pp60src." Mol Cell Biol 14(3): 1680-8.
Schlaepfer, D. D., S. K. Mitra, et al. (2004). "Control of motile and invasive cell phenotypes by focal adhesion kinase." Biochim Biophys Acta 1692(2-3): 77-102.
Shih Ie, M. and R. J. Kurman (2004). "Ovarian tumorigenesis: a proposed model based on morphological and molecular genetic analysis." Am J Pathol 164(5): 1511-8.
Sieg, D. J., C. R. Hauck, et al. (1999). "Required role of focal adhesion kinase (FAK) for integrin-stimulated cell migration." J Cell Sci 112 ( Pt 16): 2677-91.
Straus, D. S., G. Pascual, et al. (2000). "15-deoxy-delta 12,14-prostaglandin J2 inhibits multiple steps in the NF-kappa B signaling pathway." Proc Natl Acad Sci U S A 97(9): 4844-9.
Takeda, K., T. Ichiki, et al. (2001). "15-Deoxy-delta 12,14-prostaglandin J2 and thiazolidinediones activate the MEK/ERK pathway through phosphatidylinositol 3-kinase in vascular smooth muscle cells." J Biol Chem 276(52): 48950-5.
Tamura, M., J. Gu, et al. (1999). "PTEN interactions with focal adhesion kinase and suppression of the extracellular matrix-dependent phosphatidylinositol 3-kinase/Akt cell survival pathway." J Biol Chem 274(29): 20693-703.
Tamura, M., J. Gu, et al. (1998). "Inhibition of cell migration, spreading, and focal adhesions by tumor suppressor PTEN." Science 280(5369): 1614-7.
Teng, D. H., R. Hu, et al. (1997). "MMAC1/PTEN mutations in primary tumor specimens and tumor cell lines." Cancer Res 57(23): 5221-5.
Tontonoz, P., S. Singer, et al. (1997). "Terminal differentiation of human liposarcoma cells induced by ligands for peroxisome proliferator-activated receptor gamma and the retinoid X receptor." Proc Natl Acad Sci U S A 94(1): 237-41.
Torres, J. and R. Pulido (2001). "The tumor suppressor PTEN is phosphorylated by the protein kinase CK2 at its C terminus. Implications for PTEN stability to proteasome-mediated degradation." J Biol Chem 276(2): 993-8.
Totsukawa, G., Y. Yamakita, et al. (2000). "Distinct roles of ROCK (Rho-kinase) and MLCK in spatial regulation of MLC phosphorylation for assembly of stress fibers and focal adhesions in 3T3 fibroblasts." J Cell Biol 150(4): 797-806.
Tremblay, L., W. Hauck, et al. (1996). "Focal adhesion kinase (pp125FAK) expression, activation and association with paxillin and p50CSK in human metastatic prostate carcinoma." Int J Cancer 68(2): 164-71.
Valiente, M., A. Andres-Pons, et al. (2005). "Binding of PTEN to specific PDZ domains contributes to PTEN protein stability and phosphorylation by microtubule-associated serine/threonine kinases." J Biol Chem 280(32): 28936-43.
Vazquez, F., S. R. Grossman, et al. (2001). "Phosphorylation of the PTEN tail acts as an inhibitory switch by preventing its recruitment into a protein complex." J Biol Chem 276(52): 48627-30.
Vazquez, F., S. Ramaswamy, et al. (2000). "Phosphorylation of the PTEN tail regulates protein stability and function." Mol Cell Biol 20(14): 5010-8.
Wang, M., S. C. Wise, et al. (1999). "Troglitazone, an antidiabetic agent, inhibits cholesterol biosynthesis through a mechanism independent of peroxisome proliferator-activated receptor-gamma." Diabetes 48(2): 254-60.
Weng, J. R., C. Y. Chen, et al. (2006). "Beyond peroxisome proliferator-activated receptor gamma signaling: the multi-facets of the antitumor effect of thiazolidinediones." Endocr Relat Cancer 13(2): 401-13.
Westhoff, M. A., B. Serrels, et al. (2004). "SRC-mediated phosphorylation of focal adhesion kinase couples actin and adhesion dynamics to survival signaling." Mol Cell Biol 24(18): 8113-33.
Yamada, K. M. and M. Araki (2001). "Tumor suppressor PTEN: modulator of cell signaling, growth, migration and apoptosis." J Cell Sci 114(Pt 13): 2375-82.
Yang, Y. C., Y. P. Tsao, et al. (2007). "Peroxisome proliferator-activated receptor-gamma agonists cause growth arrest and apoptosis in human ovarian carcinoma cell lines." Int J Gynecol Cancer 17(2): 418-25.
Yu, D. H., C. K. Qu, et al. (1998). "Protein-tyrosine phosphatase Shp-2 regulates cell spreading, migration, and focal adhesion." J Biol Chem 273(33): 21125-31.
Zhang, S. Q., W. Yang, et al. (2004). "Shp2 regulates SRC family kinase activity and Ras/Erk activation by controlling Csk recruitment." Mol Cell 13(3): 341-55.
Zhang, W., N. Wu, et al. (2006). "PPARgamma activator rosiglitazone inhibits cell migration via upregulation of PTEN in human hepatocarcinoma cell line BEL-7404." Cancer Biol Ther 5(8): 1008-14.