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研究生:鄭成富
研究生(外文):Chen-Fu
論文名稱:RhoA/ROK路徑對乳腺細胞胰島素訊息傳遞之影響
論文名稱(外文):Effect of RhoA/ROK pathway on insulin signaling in mammary epithelial cells
指導教授:李宜儒李宜儒引用關係
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:免疫學研究所
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:2007
畢業學年度:95
語文別:中文
論文頁數:36
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對乳腺上皮細胞而言,胰島素能夠加強泌乳激素誘導泌乳基因的表現並促使細胞的存活。這些影響大部分發生在培養在基底膜的細胞,主要是因為這些細胞能夠傳遞胰島素的訊息。在本篇研究中我們進一步的發現基底膜影響乳腺細胞的胰島素訊息傳遞機制。我們發現當乳腺細胞培養在塑膠盤上是不利於胰島素訊息傳遞,並且這些細胞會表現出較高的RhoA活性以及較低的p190RhoGAP酪胺酸磷酸化現象。當乳腺細胞表現過量的dominant negative RhoA 來降低乳腺細胞的RhoA活性時,可以促進塑膠盤上細胞胰島素誘發IRS-1酪胺酸磷酸化的現象。相反地,當我們利用constitutively active RhoA提高基底膜上乳腺細胞的RhoA活性時,可以發現胰島素誘發IRS-1酪胺酸磷酸化現象被抑制。此外,當培養在塑膠盤上的乳腺細胞處理ROK抑制劑 Y27632時,IRS-1酪胺酸磷酸化表現會增加。這些實驗結果一致地指出ROK與IRS-1的交互作用。因此,RhoA/ROK路徑在乳腺上皮細胞中扮演著負調控胰島素訊息傳遞的角色。

In cultures of mammary epithelial cells, insulin potentiates prolactin-induced milk gene expression and promotes cell survival. However, these effects occur mainly in cells cultured on basement membrane due to the permissiveness of insulin signaling in these cells. Here we further explore the underlying mechanism for it. We found that cells cultured on plastic, which are inefficient in insulin signaling, exhibit higher level of RhoA activity and lower extent of tyrosine phosphorylation of p190RhoGAP. Overexpression of dominant negative RhoA in these cells augments insulin-induced tyrosine phosphorylation of IRS-1, but this signaling event is inhibited in cells cultured on basement membrane when constitutively active RhoA is expressed. Furthermore, treatment of Y27632, an inhibitor of Rho kinase (ROK), enhances tyrosine phosphorylation IRS-1 in cells cultured on plastic. Consistent with this result, greater amount of ROK is associated with IRS-1. Thus, the RhoA/ROK pathway plays a negative role in insulin signaling in mammary epithelial cells.

中文摘要 i
英文摘要 ii
縮寫檢索表 iii
第一章 緒論 1
一、 乳腺發育及其生理 1
二、 細胞外基質 1
三、 Rho家族蛋白 3
四、 胰島素 5
五、 基底膜對乳腺細胞的胰島素訊息傳遞的影響 6
六、 實驗動機與目的 7
第二章 材料方法 8
一、 實驗試劑 8
二、 懷孕小鼠 8
三、 乳腺上皮細胞的製備 9
四、 腺病毒感染 9
五、 製備細胞溶胞產物 10
六、 硫酸十二酯鈉聚丙烯醯氨板膠電泳及Commassie blue染色法 11
七、 西方墨點法 12
八、 免疫沉澱法 13
九、 Rho activity assay 13
第三章 結果 14
一、 培養在塑膠盤上的乳腺細胞具有較高的RhoA活性 14
二、 活化RhoA抑制胰島素的訊息傳遞 16
三、 表現dominant negative RhoA提高胰島素的訊息傳遞 17
四、 Y27632促進培養在塑膠盤上乳腺細胞的胰島素訊息傳遞 18
第四章 討論 20
第五章 參考文獻 32


1.McBryan, J., J. Howlin, P. A. Kenny, T. Shioda, and F. Martin. 2007. ERalpha-CITED1 co-regulated genes expressed during pubertal mammary gland development: implications for breast cancer prognosis. Oncogene.
2.Farmer, C., and M. F. Palin. 2005. Exogenous prolactin stimulates mammary development and alters expression of prolactin-related genes in prepubertal gilts. Journal of animal science 83:825-832.
3.Schenk, S., E. Hintermann, M. Bilban, N. Koshikawa, C. Hojilla, R. Khokha, and V. Quaranta. 2003. Binding to EGF receptor of a laminin-5 EGF-like fragment liberated during MMP-dependent mammary gland involution. The Journal of cell biology 161:197-209.
4.Holland, M. S., J. A. Stasko, and R. E. Holland. 2007. Influence of extracellular matrix on bovine mammary gland progenitor cell growth and differentiation. American journal of veterinary research 68:476-482.
5.Schatzmann, F., R. Marlow, and C. H. Streuli. 2003. Integrin signaling and mammary cell function. Journal of mammary gland biology and neoplasia 8:395-408.
6.Damsky, C. H., and D. Ilic. 2002. Integrin signaling: it''s where the action is. Current opinion in cell biology 14:594-602.
7.Ridley, A. J. 2001. Rho family proteins: coordinating cell responses. Trends in cell biology 11:471-477.
8.Kimura, K., M. Ito, M. Amano, K. Chihara, Y. Fukata, M. Nakafuku, B. Yamamori, J. Feng, T. Nakano, K. Okawa, A. Iwamatsu, and K. Kaibuchi. 1996. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase). Science 273:245-248.
9.Burridge, K. 1999. Crosstalk between Rac and Rho. Science 283:2028-2029.
10.Braga, V. M., A. Del Maschio, L. Machesky, and E. Dejana. 1999. Regulation of cadherin function by Rho and Rac: modulation by junction maturation and cellular context. Molecular biology of the cell 10:9-22.
11.Braga, V. M., L. M. Machesky, A. Hall, and N. A. Hotchin. 1997. The small GTPases Rho and Rac are required for the establishment of cadherin-dependent cell-cell contacts. The Journal of cell biology 137:1421-1431.
12.Hall, A. 1998. Rho GTPases and the actin cytoskeleton. Science 279:509-514.
13.Eisen, R., D. R. Ratcliffe, and G. K. Ojakian. 2004. Modulation of epithelial tubule formation by Rho kinase. American journal of physiology 286:C857-866.
14.Fujisawa, K., P. Madaule, T. Ishizaki, G. Watanabe, H. Bito, Y. Saito, A. Hall, and S. Narumiya. 1998. Different regions of Rho determine Rho-selective binding of different classes of Rho target molecules. The Journal of biological chemistry 273:18943-18949.
15.Amano, M., M. Ito, K. Kimura, Y. Fukata, K. Chihara, T. Nakano, Y. Matsuura, and K. Kaibuchi. 1996. Phosphorylation and activation of myosin by Rho-associated kinase (Rho-kinase). The Journal of biological chemistry 271:20246-20249.
16.Feng, J., M. Ito, K. Ichikawa, N. Isaka, M. Nishikawa, D. J. Hartshorne, and T. Nakano. 1999. Inhibitory phosphorylation site for Rho-associated kinase on smooth muscle myosin phosphatase. The Journal of biological chemistry 274:37385-37390.
17.Feng, J., M. Ito, Y. Kureishi, K. Ichikawa, M. Amano, N. Isaka, K. Okawa, A. Iwamatsu, K. Kaibuchi, D. J. Hartshorne, and T. Nakano. 1999. Rho-associated kinase of chicken gizzard smooth muscle. The Journal of biological chemistry 274:3744-3752.
18.Kitazawa, T., M. Eto, T. P. Woodsome, and D. L. Brautigan. 2000. Agonists trigger G protein-mediated activation of the CPI-17 inhibitor phosphoprotein of myosin light chain phosphatase to enhance vascular smooth muscle contractility. The Journal of biological chemistry 275:9897-9900.
19.Kaneko, T., M. Amano, A. Maeda, H. Goto, K. Takahashi, M. Ito, and K. Kaibuchi. 2000. Identification of calponin as a novel substrate of Rho-kinase. Biochemical and biophysical research communications 273:110-116.
20.Maekawa, M., T. Ishizaki, S. Boku, N. Watanabe, A. Fujita, A. Iwamatsu, T. Obinata, K. Ohashi, K. Mizuno, and S. Narumiya. 1999. Signaling from Rho to the actin cytoskeleton through protein kinases ROCK and LIM-kinase. Science 285:895-898.
21.Matsui, T., M. Maeda, Y. Doi, S. Yonemura, M. Amano, K. Kaibuchi, S. Tsukita, and S. Tsukita. 1998. Rho-kinase phosphorylates COOH-terminal threonines of ezrin/radixin/moesin (ERM) proteins and regulates their head-to-tail association. The Journal of cell biology 140:647-657.
22.Fukata, Y., N. Oshiro, N. Kinoshita, Y. Kawano, Y. Matsuoka, V. Bennett, Y. Matsuura, and K. Kaibuchi. 1999. Phosphorylation of adducin by Rho-kinase plays a crucial role in cell motility. The Journal of cell biology 145:347-361.
23.Tominaga, T., T. Ishizaki, S. Narumiya, and D. L. Barber. 1998. p160ROCK mediates RhoA activation of Na-H exchange. The EMBO journal 17:4712-4722.
24.Hagerty, L., D. H. Weitzel, J. Chambers, C. N. Fortner, M. H. Brush, D. Loiselle, H. Hosoya, and T. A. Haystead. 2007. ROCK1 phosphorylates and activates zipper-interacting protein kinase. The Journal of biological chemistry 282:4884-4893.
25.Lim, M. J., K. J. Choi, Y. Ding, J. H. Kim, B. S. Kim, Y. H. Kim, J. Lee, W. Choe, I. Kang, J. Ha, K. S. Yoon, and S. S. Kim. 2007. RhoA/Rho Kinase Blocks Muscle Differentiation via Serine Phosphorylation of Insulin Receptor Substrate-1 and 2. Mol Endocrinol.
26.Ahren, B., and G. Pacini. 2004. Importance of quantifying insulin secretion in relation to insulin sensitivity to accurately assess beta cell function in clinical studies. European journal of endocrinology 150:97-104.
27.Zeitz, U., K. Weber, D. W. Soegiarto, E. Wolf, R. Balling, and R. G. Erben. 2003. Impaired insulin secretory capacity in mice lacking a functional vitamin D receptor. FASEB J 17:509-511.
28.Shen, H. Q., M. D. Roth, and R. G. Peterson. 1998. The effect of glucose and glucagon-like peptide-1 stimulation on insulin release in the perfused pancreas in a non-insulin-dependent diabetes mellitus animal model. Metabolism: clinical and experimental 47:1042-1047.
29.Okamoto, H. 1999. The Reg gene family and Reg proteins: with special attention to the regeneration of pancreatic beta-cells. Journal of hepato-biliary-pancreatic surgery 6:254-262.
30.Okamoto, H., and S. Takasawa. 2002. Recent advances in the Okamoto model: the CD38-cyclic ADP-ribose signal system and the regenerating gene protein (Reg)-Reg receptor system in beta-cells. Diabetes 51 Suppl 3:S462-473.
31.Fehmann, H. C., J. Jiang, D. Pitt, J. Schweinfurth, and B. Goke. 1996. Ligand-induced regulation of glucagon-like peptide-I receptor function and expression in insulin-secreting beta cells. Pancreas 13:273-282.
32.Biddinger, S. B., and C. R. Kahn. 2006. From mice to men: insights into the insulin resistance syndromes. Annual review of physiology 68:123-158.
33.White, M. F., and C. R. Kahn. 1994. The insulin signaling system. The Journal of biological chemistry 269:1-4.
34.Sun, X. J., M. Miralpeix, M. G. Myers, Jr., E. M. Glasheen, J. M. Backer, C. R. Kahn, and M. F. White. 1992. Expression and function of IRS-1 in insulin signal transmission. The Journal of biological chemistry 267:22662-22672.
35.Araki, E., M. A. Lipes, M. E. Patti, J. C. Bruning, B. Haag, 3rd, R. S. Johnson, and C. R. Kahn. 1994. Alternative pathway of insulin signalling in mice with targeted disruption of the IRS-1 gene. Nature 372:186-190.
36.Choi, K. M., I. Barash, and R. E. Rhoads. 2004. Insulin and prolactin synergistically stimulate beta-casein messenger ribonucleic acid translation by cytoplasmic polyadenylation. Mol Endocrinol 18:1670-1686.
37.Paulsson, M. 1992. Basement membrane proteins: structure, assembly, and cellular interactions. Critical reviews in biochemistry and molecular biology 27:93-127.
38.Farrelly, N., Y. J. Lee, J. Oliver, C. Dive, and C. H. Streuli. 1999. Extracellular matrix regulates apoptosis in mammary epithelium through a control on insulin signaling. The Journal of cell biology 144:1337-1348.
39.Molnar, G., M. C. Dagher, M. Geiszt, J. Settleman, and E. Ligeti. 2001. Role of prenylation in the interaction of Rho-family small GTPases with GTPase activating proteins. Biochemistry 40:10542-10549.
40.Chiarugi, P., M. L. Taddei, P. Cirri, D. Talini, F. Buricchi, G. Camici, G. Manao, G. Raugei, and G. Ramponi. 2000. Low molecular weight protein-tyrosine phosphatase controls the rate and the strength of NIH-3T3 cells adhesion through its phosphorylation on tyrosine 131 or 132. The Journal of biological chemistry 275:37619-37627.
41.Heneberg, P., and P. Draber. 2002. Nonreceptor protein tyrosine and lipid phosphatases in type I fc(epsilon) receptor-mediated activation of mast cells and basophils. International archives of allergy and immunology 128:253-263.
42.Nimnual, A. S., L. J. Taylor, and D. Bar-Sagi. 2003. Redox-dependent downregulation of Rho by Rac. Nature cell biology 5:236-241.
43.Gu, J., Y. Sumida, N. Sanzen, and K. Sekiguchi. 2001. Laminin-10/11 and fibronectin differentially regulate integrin-dependent Rho and Rac activation via p130(Cas)-CrkII-DOCK180 pathway. The Journal of biological chemistry 276:27090-27097.
44.Akhtar, N., and C. H. Streuli. 2006. Rac1 links integrin-mediated adhesion to the control of lactational differentiation in mammary epithelia. The Journal of cell biology 173:781-793.
45.Shi, J., and L. Wei. 2007. Rho kinase in the regulation of cell death and survival. Archivum immunologiae et therapiae experimentalis 55:61-75.
46.Furukawa, N., P. Ongusaha, W. J. Jahng, K. Araki, C. S. Choi, H. J. Kim, Y. H. Lee, K. Kaibuchi, B. B. Kahn, H. Masuzaki, J. K. Kim, S. W. Lee, and Y. B. Kim. 2005. Role of Rho-kinase in regulation of insulin action and glucose homeostasis. Cell metabolism 2:119-129.
47.Eisen, R., S. Walid, D. R. Ratcliffe, and G. K. Ojakian. 2006. Regulation of epithelial tubule formation by Rho family GTPases. American journal of physiology 290:C1297-1309.
48.Wozniak, M. A., R. Desai, P. A. Solski, C. J. Der, and P. J. Keely. 2003. ROCK-generated contractility regulates breast epithelial cell differentiation in response to the physical properties of a three-dimensional collagen matrix. The Journal of cell biology 163:583-595.
49.Marshman, E., and C. H. Streuli. 2002. Insulin-like growth factors and insulin-like growth factor binding proteins in mammary gland function. Breast Cancer Res 4:231-239.
50.Kanda, T., S. Wakino, K. Homma, K. Yoshioka, S. Tatematsu, K. Hasegawa, I. Takamatsu, N. Sugano, K. Hayashi, and T. Saruta. 2006. Rho-kinase as a molecular target for insulin resistance and hypertension. Faseb J 20:169-171.
51.Begum, N., O. A. Sandu, M. Ito, S. M. Lohmann, and A. Smolenski. 2002. Active Rho kinase (ROK-alpha ) associates with insulin receptor substrate-1 and inhibits insulin signaling in vascular smooth muscle cells. The Journal of biological chemistry 277:6214-6222.




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