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研究生:謝佳筑
研究生(外文):Chia-Chu Hsieh
論文名稱:探討哺乳類動物細胞中HURP基因表現之調控機制
論文名稱(外文):study the regulation of HURP gene expression in mammalian cells
指導教授:周成功
指導教授(外文):Chen-Kung Chou
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:55
中文關鍵詞:肝癌過量表現蛋白啟動子訊號傳導
外文關鍵詞:hepatoma-up regulated proteinpromotersignal transductionMLK3JNK
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肝癌是世界上盛行率極高的惡性腫瘤,尤其是亞洲地區癌症的頭號殺手。本實驗室楊鉅文學長利用微陣列及EST資料庫的分析,找出一個在肝癌過量表現且與細胞週期相關的新穎基因,並命為HURP (Hepatoma Up Regulated Protein)基因。已知HURP的穩定轉殖細胞株具有低血清之需求及在不須貼附下即可生長等癌細胞特性。故HURP基因表現之調控機制為探討肝癌致病機轉之重要方針。
由實驗結果得知HURP最小啟動子位於-55~+86區域。我們進一步利用生物資訊學分析啟動子可能調控HURP基因表現的轉錄因子結合序列,發現可能存在與細胞週期調控有關的轉錄因子YB-1 (Y-box-binding protein)的結合序列,進一步亦發現隨著HuH7細胞中YB-1的表現量增加,HURP 啟動子活性越低。
另一方面,我們發現MLK3 (Mixed lineage kinase 3) 可以抑制HURP之啟動子活性及其基因表現,且隨著MLK3表現量增加,此抑制現象越顯著。且MLK3對HURP基因表現之抑制現象可能與其激酶活性有關。但對HURP啟動子CCAAT boxes加以突變後,發現MLK3無法抑制此突變啟動子活性。
因大量表現HuH7 細胞中不具激酶活性之MLK3會降低TGF-β對HURP基因表現之抑制現象,故此抑制現象與MLK3之激酶活性有關。隨著ceramide濃度增加,HEK 293T 細胞中HURP基因表現逐漸減少,推論ceramide可能是藉由活化MLK3或其他蛋白而降低HURP基因表現。實驗結果發現大量表現HEK 293T細胞中JIP可阻礙MLK3對HURP基因表現之抑制現象。這些結果顯示MLK3可能經由活化JNK訊號途徑而抑制HURP基因的表現。
Hepatocellular carcinoma (HCC) is one of the most common human malignancies worldwide, and it causes a serious public health problem in Asian population. Based on microarray and EST database analysis we have identified a novel cell cycle regulated gene from HCC, named Hepatoma Up Regulated Protein (HURP). HURP-stable transfectants have the characteristics of a reduced serum dependency and exhibit anchorage-independent cell growth. Therefore, study of how the expression is regulated becomes an important direction to understanding the carcinogenesis of HCC.
We characterized that the minimal HURP promoter region of HURP was resided within –55 to +86. By the bioinformatics analysis, we furthermore identified two possible regulatory cis elements, Y-box-binding protein (YB-1) binding sites, of HURP promoter. However, we showed that YB-1 repressed HURP promoter activity in a dose-dependent manner in HuH7 cells.
In addition, we found that MLK3 (Mixed-Lineage Kinase 3) inhibited the promoter activity and the gene expression of HURP in a dose-dependent manner. The inhibitory effect of MLK3 on HURP gene expression was kinase activity dependent. We also found that mutations of CCAAT boxes in HURP promoter region impaired the MLK3-mediated down-regulation of HURP promoter activity.
In view of that over-expression of kinase dead MLK3 mutant could
prevent TGF-β-mediated suppression of HURP gene expression in HuH7, MLK3 might be involved in TGF-β-mediated suppression of HURP gene expression. Ceramide, an activator of MLK3, also suppressed HURP gene expression through MLK3 activation or other protein in HEK 293T and HuH7 cells. Our results indicated that the MLK3-mediated suppression of HURP gene expression could be reversed in the presence of JIP (JNK-interacting protein), a JNK specific inhibitor. Taken together, these results suggested MLK3 might act through JNK signaling pathway specifically to inhibit HURP gene expression.
References
Bargou, R. C., Jurchott, K., Wagener, C., Bergmann, S., Metzner, S., Bommert, K., Mapara, M. Y., Winzer, K. J., Dietel, M., Dorken, B., and Royer, H. D. (1997). Nuclear localization and increased levels of transcription factor YB-1 in primary human breast cancers are associated with intrinsic MDR1 gene expression. Nat Med 3, 447-450.
Bourbon, N. A., Yun, J., and Kester, M. (2000). Ceramide directly activates protein kinase C zeta to regulate a stress-activated protein kinase signaling complex. J Biol Chem 275, 35617-35623.
Chang, L., and Karin, M. (2001). Mammalian MAP kinase signalling cascades. Nature 410, 37-40.
Davis, R. J. (2000). Signal transduction by the JNK group of MAP kinases. Cell 103, 239-252.
Eferl, R., Ricci, R., Kenner, L., Zenz, R., David, J. P., Rath, M., and Wagner, E. F. (2003). Liver tumor development. c-Jun antagonizes the proapoptotic activity of p53. Cell 112, 181-192.
Eferl, R., and Wagner, E. F. (2003). AP-1: a double-edged sword in tumorigenesis. Nat Rev Cancer 3, 859-868.
English, J. M., and Cobb, M. H. (2002). Pharmacological inhibitors of MAPK pathways. Trends Pharmacol Sci 23, 40-45.
Gallo, K. A., and Johnson, G. L. (2002). Mixed-lineage kinase control of JNK and p38 MAPK pathways. Nat Rev Mol Cell Biol 3, 663-672.
Hipfel, R., Schittek, B., Bodingbauer, Y., and Garbe, C. (2000). Specifically regulated genes in malignant melanoma tissues identified by subtractive hybridization. Br J Cancer 82, 1149-1157.
Hlavacek, W. S., Faeder, J. R., Blinov, M. L., Perelson, A. S., and Goldstein, B. (2003). The complexity of complexes in signal transduction. Biotechnol Bioeng 84, 783-794.
Hamilton, S.R. and Aaltonen, L.A.(2000). Pathology&Genetics: Tumors of the dogestive System(Lyon:IARC)
Holm, P. S., Bergmann, S., Jurchott, K., Lage, H., Brand, K., Ladhoff, A., Mantwill, K., Curiel, D. T., Dobbelstein, M., Dietel, M., et al. (2002). YB-1 relocates to the nucleus in adenovirus-infected cells and facilitates viral replication by inducing E2 gene expression through the E2 late promoter. J Biol Chem 277, 10427-10434.
Hunter, T. (2000). Signaling--2000 and beyond. Cell 100, 113-127.
Iyer, V. R., Eisen, M. B., Ross, D. T., Schuler, G., Moore, T., Lee, J. C., Trent, J. M., Staudt, L. M., Hudson, J., Jr., Boguski, M. S., et al. (1999). The transcriptional program in the response of human fibroblasts to serum. Science 283, 83-87.
Jurchott, K., Bergmann, S., Stein, U., Walther, W., Janz, M., Manni, I., Piaggio, G., Fietze, E., Dietel, M., and Royer, H. D. (2003). YB-1 as a cell cycle-regulated transcription factor facilitating cyclin A and cyclin B1 gene expression. J Biol Chem 278, 27988-27996.
Kim, K. Y., Kim, B. C., Xu, Z., and Kim, S. J. (2004). Mixed lineage kinase (MLK) 3-activated p38 MAP kinase mediates TGF--beta-induced apoptosis in hepatoma cells. J Biol Chem.
Kohno, K., Izumi, H., Uchiumi, T., Ashizuka, M., and Kuwano, M. (2003). The pleiotropic functions of the Y-box-binding protein, YB-1. Bioessays 25, 691-698.
Kyriakis, J. M., and Avruch, J. (2001). Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev 81, 807-869.
Ladomery, M., and Sommerville, J. (1995). A role for Y-box proteins in cell proliferation. Bioessays 17, 9-11.
Lasham, A., Lindridge, E., Rudert, F., Onrust, R., and Watson, J. (2000). Regulation of the human fas promoter by YB-1, Puralpha and AP-1 transcription factors. Gene 252, 1-13.
Lasham, A., Moloney, S., Hale, T., Homer, C., Zhang, Y. F., Murison, J. G., Braithwaite, A. W., and Watson, J. (2003). The Y-box-binding protein, YB1, is a potential negative regulator of the p53 tumor suppressor. J Biol Chem 278, 35516-35523.
Martinez, J.D., Parker, M.T., Fultz, K.E., Ignatenko, N.A., and Gerner, E.W. (2003) Burger's Medicinal Chemistry and drug Discovery, Six Edition, Volume 5; Chemotherapeutic Agents
Mathias, S., Pena, L. A., and Kolesnick, R. N. (1998). Signal transduction of stress via ceramide. Biochem J 335 ( Pt 3), 465-480.
Matsumoto, K., and Wolffe, A. P. (1998). Gene regulation by Y-box proteins: coupling control of transcription and translation. Trends Cell Biol 8, 318-323.
Merritt, S. E., Mata, M., Nihalani, D., Zhu, C., Hu, X., and Holzman, L. B. (1999). The mixed lineage kinase DLK utilizes MKK7 and not MKK4 as substrate. J Biol Chem 274, 10195-10202.
Muller, G., Storz, P., Bourteele, S., Doppler, H., Pfizenmaier, K., Mischak, H., Philipp, A., Kaiser, C., and Kolch, W. (1998). Regulation of Raf-1 kinase by TNF via its second messenger ceramide and cross-talk with mitogenic signalling. Embo J 17, 732-742.
Murray, C. J., and Lopez, A. D. (1997). Mortality by cause for eight regions of the world: Global Burden of Disease Study. Lancet 349, 1269-1276.
Nihalani, D., Meyer, D., Pajni, S., and Holzman, L. B. (2001). Mixed lineage kinase-dependent JNK activation is governed by interactions of scaffold protein JIP with MAPK module components. Embo J 20, 3447-3458.
Nojima, H. (1997). Cell cycle checkpoints, chromosome stability and the progression of cancer. Hum Cell 10, 221-230.
Oda, Y., Sakamoto, A., Shinohara, N., Ohga, T., Uchiumi, T., Kohno, K., Tsuneyoshi, M., Kuwano, M., and Iwamoto, Y. (1998). Nuclear expression of YB-1 protein correlates with P-glycoprotein expression in human osteosarcoma. Clin Cancer Res 4, 2273-2277.
Ohga, T., Uchiumi, T., Makino, Y., Koike, K., Wada, M., Kuwano, M., and Kohno, K. (1998). Direct involvement of the Y-box binding protein YB-1 in genotoxic stress-induced activation of the human multidrug resistance 1 gene. J Biol Chem 273, 5997-6000.
Okamoto, T., Izumi, H., Imamura, T., Takano, H., Ise, T., Uchiumi, T., Kuwano, M., and Kohno, K. (2000). Direct interaction of p53 with the Y-box binding protein, YB-1: a mechanism for regulation of human gene expression. Oncogene 19, 6194-6202.
Passegue, E., Jochum, W., Schorpp-Kistner, M., Mohle-Steinlein, U., and Wagner, E. F. (2001). Chronic myeloid leukemia with increased granulocyte progenitors in mice lacking junB expression in the myeloid lineage. Cell 104, 21-32.
Rosenberg, A. R., Zindy, F., Le Deist, F., Mouly, H., Metezeau, P., Brechot, C., and Lamas, E. (1995). Overexpression of human cyclin A advances entry into S phase. Oncogene 10, 1501-1509.
Sathyanarayana, P., Barthwal, M. K., Kundu, C. N., Lane, M. E., Bergmann, A., Tzivion, G., and Rana, A. (2002). Activation of the Drosophila MLK by ceramide reveals TNF-alpha and ceramide as agonists of mammalian MLK3. Mol Cell 10, 1527-1533.
Shibahara, K., Sugio, K., Osaki, T., Uchiumi, T., Maehara, Y., Kohno, K., Yasumoto, K., Sugimachi, K., and Kuwano, M. (2001). Nuclear expression of the Y-box binding protein, YB-1, as a novel marker of disease progression in non-small cell lung cancer. Clin Cancer Res 7, 3151-3155.
Shibao, K., Takano, H., Nakayama, Y., Okazaki, K., Nagata, N., Izumi, H., Uchiumi, T., Kuwano, M., Kohno, K., and Itoh, H. (1999). Enhanced coexpression of YB-1 and DNA topoisomerase II alpha genes in human colorectal carcinomas. Int J Cancer 83, 732-737.
Smale, S. T., and Fisher, A. G. (2002). Chromatin structure and gene regulation in the immune system. Annu Rev Immunol 20, 427-462.
Staib, F., Hussain, S. P., Hofseth, L. J., Wang, X. W., and Harris, C. C. (2003). TP53 and liver carcinogenesis. Hum Mutat 21, 201-216.
Swamynathan, S. K., Nambiar, A., and Guntaka, R. V. (1998). Role of single-stranded DNA regions and Y-box proteins in transcriptional regulation of viral and cellular genes. Faseb J 12, 515-522.
Teramoto, H., Coso, O. A., Miyata, H., Igishi, T., Miki, T., and Gutkind, J. S. (1996). Signaling from the small GTP-binding proteins Rac1 and Cdc42 to the c-Jun N-terminal kinase/stress-activated protein kinase pathway. A role for mixed lineage kinase 3/protein-tyrosine kinase 1, a novel member of the mixed lineage kinase family. J Biol Chem 271, 27225-27228.
Tsou, A. P., Yang, C. W., Huang, C. Y., Yu, R. C., Lee, Y. C., Chang, C. W., Chen, B. R., Chung, Y. F., Fann, M. J., Chi, C. W., et al. (2003). Identification of a novel cell cycle regulated gene, HURP, overexpressed in human hepatocellular carcinoma. Oncogene 22, 298-307.
Visintin, R., Prinz, S., and Amon, A. (1997). CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis. Science 278, 460-463.
Vogt, P. K. (2002). Fortuitous convergences: the beginnings of JUN. Nat Rev Cancer 2, 465-469.
Zhang, H. S., Gavin, M., Dahiya, A., Postigo, A. A., Ma, D., Luo, R. X., Harbour, J. W., and Dean, D. C. (2000). Exit from G1 and S phase of the cell cycle is regulated by repressor complexes containing HDAC-Rb-hSWI/SNF and Rb-hSWI/SNF. Cell 101, 79-89.
Zhang, M. Q. (1998). Identification of human gene core promoters in silico. Genome Res 8, 319-326.
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