跳到主要內容

臺灣博碩士論文加值系統

(3.236.110.106) 您好!臺灣時間:2021/07/27 18:57
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:郭怡伶
研究生(外文):Yi-Ling Kuo
論文名稱:CD74在癌症轉移中所扮演的角色
論文名稱(外文):Potential Role of CD74 in Cancer Metastasis
指導教授:呂思潔呂思潔引用關係
指導教授(外文):Sy-Jye Leu
學位類別:碩士
校院名稱:臺北醫學大學
系所名稱:細胞及分子生物研究所
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:39
中文關鍵詞:癌症轉移
外文關鍵詞:CD74metastasis
相關次數:
  • 被引用被引用:0
  • 點閱點閱:121
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
CD74為major histocompatibility complex class II invariant chain,是第二類非多形性穿膜的醣化蛋白,在抗原呈現的過程中為MHC class II的護衛蛋白。近年來的研究發現CD74亦為一個傳訊分子,可以傳遞使細胞增生存活的訊息。最近也證實了CD74除了表現在抗原呈現細胞上以外,在許多的肉瘤上也都有CD74的表現,像是腎癌、肺癌、胃癌及胸腺來源的肉瘤等等。但是,對於CD74和腫瘤惡化的關係目前卻不是很清楚。
利用subtractive hybridization的方式比對肺癌轉移及原位肺癌病人的cDNA library發現,CD74的基因表現與肺癌的轉移有關。首先我們利用免疫組織染色觀察肺癌病人的組織切片確認了在腫瘤細胞相對於鄰近的正常細胞CD74的確有過度表現的現象。但奇怪的是,在15株肺部相關的細胞株(包含正常及肺癌細胞株)中,利用RT-PCR及Western blotting都沒有觀察到CD74的表現。我們利用PCR的方法以及用Western blotting分析處理過甲基轉移脢抑制劑的細胞株來確認這些細胞中的CD74基因沒有被剔除,而其mRNA的不表現也不是因為DNA被甲基化的關係。同時,我們建構了兩個CD74的isoforms-p33及p35,並利用兩株不同侵襲能力的肺癌細胞株—低侵襲力的CL1-0細胞及高侵襲力的CL1-5-F4細胞來作為細胞模式。將CD74 p33過度表現在CL1-5-F4細胞中可以更加強細胞的爬行和侵襲能力,但在CL1-0則沒有觀察到這個現象。而過度表現CD74 p35則不論在CL1-0或CL1-5-F4細胞中對於細胞的爬行、侵襲以及生長能力都沒有影響。我們推測,CD74可能是一個協同因子能夠幫助惡性腫瘤的惡化。
CD74 (major histocompatibility complex class II invariant chain) is a type II transmembrane protein which was thought to function mainly as an MHC class II chaperone during antigen presentation. Recently, CD74 was reported to have a role as a signaling molecule for cell survival. In addition to its expression on antigen-presenting cells, it is expressed by sarcomas of renal, lung, gastric, and thymic origin and by certin sarcomas. However, little is known for the relationship in CD74 and tumor malignancy.
CD74 gene is originally cloned out from a cDNA library by subtracting metastatic tumors from primary tumors of a patient with lung adenocarcinoma. Our immunohistochemical study confirmed its overexpression on the tumor parts of patients with lung cancer as compared to the tumor-adjacent normal counterparts. Intriguingly, no expression of CD74 was observed in 15 lung cancer or normal-like cell lines by RT-PCR and Western blotting analysis. We demonstrated that absence of CD74 mRNA or protein expression is not due to gene deletion or DNA methylation through PCR and Western blotting analysis of 5-aza-2’-deoxycytidine (inhibitor of DNA methyltransferase) – treated cells. We constructed two isoforms of CD74, p33 and p35, and use low-/ high-invasive lung cancer cell line, CL1-0 and CL1-5-F4, as cell model. Overexpression of CD74 p33 in a high-invasive lung cancer cell line, CL1-5-F4, could further increase its migration and invasion ability, but the phenomenon was not observed in low-invasive lung cancer cell line, CL1-0. However, overexpression of CD74 p35 in both CL1-0 and CL1-5-F4 cells did not affect cell migration, invasion, and cell growth. Therefore, these data suggested that CD74 may play a co-factor which promote tumor malignancy.
目 次
中文摘要.....................第 i頁
英文摘要........................第 ii頁
圖目錄...........................第 iii頁

壹、緒論....................第1頁
貳、實驗目的..................第4頁
參、研究材料與方法
ㄧ、免疫組織染色(Immunohistochemistry)...第 5頁
二、細胞培養(Cell culture)..........第 6頁
三、反轉錄聚合脢鏈鎖反應(RT-PCR).......第 7頁
四、西方墨點法(Western blotting).......第 7頁
五、以DNA甲基轉移脢抑制劑-5-aza-2’-deoxycytidine處理細胞....................第 8頁
六、CD74基因構築(CD74 gene construction).第 9頁
七、流式細胞儀分析細胞表面蛋白表現......第 9頁
八、細胞生長分析(Cell growth assay)...........第 10頁
九、細胞爬行與侵襲能力分析(Migration and Invasion assay).......................第 11頁
十、以recombinant human MIF刺激細胞.........第 12頁
十一、統計分析.....................第 12頁
肆、實驗結果與分析
一、CD74在肺癌病人之腫瘤部分有過度表現的現象.....第 13頁
二、觀察CD74在低侵襲能力之細胞株CL1-0與高侵襲能力之細胞株CL1-5-F4中mRNA及蛋白質表現量之差異.......第 13頁
三、探討為何在肺癌病人的腫瘤部份可以看到明顯的CD74過度表現,在各個肺癌細胞株中卻沒有CD74蛋白的表現....... 第 14頁
四、確認構築的CD74基因之表現............. 第 15頁
五、探討在CL1-0與CL1-5-F4細胞中過度表現CD74 p33及p35後,對於其細胞生長的影響.................. 第 16頁
六、探討在CL1-0與CL1-5-F4細胞中過度表現CD74 p33及p35後,對於其細胞爬行能力的影響................ 第 17頁
七、探討在CL1-0與CL1-5-F4細胞中過度表現CD74 p33後,對於其細胞侵襲能力的影響................... 第 17頁
八、觀察已知與CD74蛋白有結合之蛋白—MIF和CD44的基因與蛋白在CL1-0與CL1-5-F4細胞中之表現............第 18頁
九、觀察在CL1-0與CL1-5-F4細胞中過度表現CD74 p33後,利用recombinant human MIF刺激後對於細胞的影響......第 19頁
伍、結論與討論......................第21頁
陸、圖表........................第24頁
柒、參考文獻......................第33頁
捌、附錄
Appendix 1. Primers of RT-PCR, PCR, and CD74 constructions...第 37頁
Appendex 2. Different patterns of splicing that yield CD74 p33 and p41 mRNA....................第 38頁
Appendex 3. Vector map of CD74 p33 and p35 construct......第 39頁
1.organization, w. h. 2007. facts about cancer http://www.who.int/mediacentre/factsheets/fs297/en/index.html.
2.行政院衛生署統計室. 2007. Health and National health Insurance Annual Statistics Information Service http://www.doh.gov.tw/statistic/index.htm. Taipei.
3.Hanahan, D., and R. A. Weinberg. 2000. The hallmarks of cancer. Cell 100:57-70.
4.Liotta, L. A., C. N. Rao, and U. M. Wewer. 1986. Biochemical interactions of tumor cells with the basement membrane. Annual review of biochemistry 55:1037-1057.
5.Strubin, M., C. Berte, and B. Mach. 1986. Alternative splicing and alternative initiation of translation explain the four forms of the Ia antigen-associated invariant chain. The EMBO journal 5:3483-3488.
6.Lotteau, V., L. Teyton, A. Peleraux, T. Nilsson, L. Karlsson, S. L. Schmid, V. Quaranta, and P. A. Peterson. 1990. Intracellular transport of class II MHC molecules directed by invariant chain. Nature 348:600-605.
7.Stumptner-Cuvelette, P., and P. Benaroch. 2002. Multiple roles of the invariant chain in MHC class II function. Biochimica et biophysica acta 1542:1-13.
8.Ong, G. L., D. M. Goldenberg, H. J. Hansen, and M. J. Mattes. 1999. Cell surface expression and metabolism of major histocompatibility complex class II invariant chain (CD74) by diverse cell lines. Immunology 98:296-302.
9.Naujokas, M. F., M. Morin, M. S. Anderson, M. Peterson, and J. Miller. 1993. The chondroitin sulfate form of invariant chain can enhance stimulation of T cell responses through interaction with CD44. Cell 74:257-268.
10.Becker-Herman, S., G. Arie, H. Medvedovsky, A. Kerem, and I. Shachar. 2005. CD74 is a member of the regulated intramembrane proteolysis-processed protein family. Molecular biology of the cell 16:5061-5069.
11.Matza, D., A. Kerem, H. Medvedovsky, F. Lantner, and I. Shachar. 2002. Invariant chain-induced B cell differentiation requires intramembrane proteolytic release of the cytosolic domain. Immunity 17:549-560.
12.Matza, D., O. Wolstein, R. Dikstein, and I. Shachar. 2001. Invariant chain induces B cell maturation by activating a TAF(II)105-NF-kappaB-dependent transcription program. The Journal of biological chemistry 276:27203-27206.
13.Starlets, D., Y. Gore, I. Binsky, M. Haran, N. Harpaz, L. Shvidel, S. Becker-Herman, A. Berrebi, and I. Shachar. 2006. Cell-surface CD74 initiates a signaling cascade leading to cell proliferation and survival. Blood 107:4807-4816.
14.Henne, C., F. Schwenk, N. Koch, and P. Moller. 1995. Surface expression of the invariant chain (CD74) is independent of concomitant expression of major histocompatibility complex class II antigens. Immunology 84:177-182.
15.Meyer-Siegler, K. L., K. A. Iczkowski, L. Leng, R. Bucala, and P. L. Vera. 2006. Inhibition of macrophage migration inhibitory factor or its receptor (CD74) attenuates growth and invasion of DU-145 prostate cancer cells. J Immunol 177:8730-8739.
16.Koide, N., T. Yamada, R. Shibata, T. Mori, M. Fukuma, K. Yamazaki, K. Aiura, M. Shimazu, S. Hirohashi, Y. Nimura, and M. Sakamoto. 2006. Establishment of perineural invasion models and analysis of gene expression revealed an invariant chain (CD74) as a possible molecule involved in perineural invasion in pancreatic cancer. Clin Cancer Res 12:2419-2426.
17.Meyer-Siegler, K. L., E. C. Leifheit, and P. L. Vera. 2004. Inhibition of macrophage migration inhibitory factor decreases proliferation and cytokine expression in bladder cancer cells. BMC cancer 4:34.
18.Ishigami, S., S. Natsugoe, K. Tokuda, A. Nakajo, H. Iwashige, K. Aridome, S. Hokita, and T. Aikou. 2001. Invariant chain expression in gastric cancer. Cancer letters 168:87-91.
19.Young, A. N., M. B. Amin, C. S. Moreno, S. D. Lim, C. Cohen, J. A. Petros, F. F. Marshall, and A. S. Neish. 2001. Expression profiling of renal epithelial neoplasms: a method for tumor classification and discovery of diagnostic molecular markers. The American journal of pathology 158:1639-1651.
20.Ioachim, H. L., S. E. Pambuccian, M. Hekimgil, F. R. Giancotti, and B. H. Dorsett. 1996. Lymphoid monoclonal antibodies reactive with lung tumors. Diagnostic applications. The American journal of surgical pathology 20:64-71.
21.Marti, G. E., V. Zenger, M. Brown, D. M. Marti, J. V. Melo, M. Crescenzi, B. Dadey, T. Han, P. Bertin, N. E. Caporaso, and et al. 1992. Antigenic expression of B-cell chronic lymphocytic leukemic cell lines. Leukemia & lymphoma 7:497-504.
22.Chu, Y. W., P. C. Yang, S. C. Yang, Y. C. Shyu, M. J. Hendrix, R. Wu, and C. W. Wu. 1997. Selection of invasive and metastatic subpopulations from a human lung adenocarcinoma cell line. American journal of respiratory cell and molecular biology 17:353-360.
23.Chen, J. J., K. Peck, T. M. Hong, S. C. Yang, Y. P. Sher, J. Y. Shih, R. Wu, J. L. Cheng, S. R. Roffler, C. W. Wu, and P. C. Yang. 2001. Global analysis of gene expression in invasion by a lung cancer model. Cancer research 61:5223-5230.
24.Stresemann, C., B. Brueckner, T. Musch, H. Stopper, and F. Lyko. 2006. Functional diversity of DNA methyltransferase inhibitors in human cancer cell lines. Cancer research 66:2794-2800.
25.Leng, L., C. N. Metz, Y. Fang, J. Xu, S. Donnelly, J. Baugh, T. Delohery, Y. Chen, R. A. Mitchell, and R. Bucala. 2003. MIF signal transduction initiated by binding to CD74. The Journal of experimental medicine 197:1467-1476.
26.Mitchell, R. A., H. Liao, J. Chesney, G. Fingerle-Rowson, J. Baugh, J. David, and R. Bucala. 2002. Macrophage migration inhibitory factor (MIF) sustains macrophage proinflammatory function by inhibiting p53: regulatory role in the innate immune response. Proceedings of the National Academy of Sciences of the United States of America 99:345-350.
27.Nishihira, J., T. Ishibashi, T. Fukushima, B. Sun, Y. Sato, and S. Todo. 2003. Macrophage migration inhibitory factor (MIF): Its potential role in tumor growth and tumor-associated angiogenesis. Annals of the New York Academy of Sciences 995:171-182.
28.Shimizu, T., R. Abe, H. Nakamura, A. Ohkawara, M. Suzuki, and J. Nishihira. 1999. High expression of macrophage migration inhibitory factor in human melanoma cells and its role in tumor cell growth and angiogenesis. Biochemical and biophysical research communications 264:751-758.
29.Shi, X., L. Leng, T. Wang, W. Wang, X. Du, J. Li, C. McDonald, Z. Chen, J. W. Murphy, E. Lolis, P. Noble, W. Knudson, and R. Bucala. 2006. CD44 is the signaling component of the macrophage migration inhibitory factor-CD74 receptor complex. Immunity 25:595-606.
30.Mitchell, R. A., C. N. Metz, T. Peng, and R. Bucala. 1999. Sustained mitogen-activated protein kinase (MAPK) and cytoplasmic phospholipase A2 activation by macrophage migration inhibitory factor (MIF). Regulatory role in cell proliferation and glucocorticoid action. The Journal of biological chemistry 274:18100-18106.
31.Saxena, N. K., D. Sharma, X. Ding, S. Lin, F. Marra, D. Merlin, and F. A. Anania. 2007. Concomitant activation of the JAK/STAT, PI3K/AKT, and ERK signaling is involved in leptin-mediated promotion of invasion and migration of hepatocellular carcinoma cells. Cancer research 67:2497-2507.
32.Tremblay, P. L., F. A. Auger, and J. Huot. 2006. Regulation of transendothelial migration of colon cancer cells by E-selectin-mediated activation of p38 and ERK MAP kinases. Oncogene 25:6563-6573.
33.Tomiyasu, M., I. Yoshino, R. Suemitsu, T. Okamoto, and K. Sugimachi. 2002. Quantification of macrophage migration inhibitory factor mRNA expression in non-small cell lung cancer tissues and its clinical significance. Clin Cancer Res 8:3755-3760.
34.Roche, P. A., C. L. Teletski, E. Stang, O. Bakke, and E. O. Long. 1993. Cell surface HLA-DR-invariant chain complexes are targeted to endosomes by rapid internalization. Proceedings of the National Academy of Sciences of the United States of America 90:8581-8585.
35.Burton, J. D., S. Ely, P. K. Reddy, R. Stein, D. V. Gold, T. M. Cardillo, and D. M. Goldenberg. 2004. CD74 is expressed by multiple myeloma and is a promising target for therapy. Clin Cancer Res 10:6606-6611.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top