中心體存在於大部份的脊髓動物之中，構成微管組成中心（microtubulin-organizing center; MTOC）。中心體對於紡綞絲的形成、細胞極性化作用、細胞的移動扮演著重要的角色。在典型的體細胞中，中心體主要是由一對中心粒所組成，周圍由稱為pericentriolar material (PCM)的物質所環繞。Human Ninein基因已經被命名為中心體相關蛋白，已知它和microtubules負端的capping, 中心粒的位置, 中心體的成熟以及作為anchoring protein有關。相關研究也指出hNinein的CCII domain與中心體的targeting signal, 調節訊號以及磷酸化作用的位置有關聯性。
在本篇論文中，我們針對human Ninein C-端四種不同splicing isoforms在特性上、表現上及功能上是否有影響與差異進行探討，包括isoform 1、isoform 2、isoform 5以及本篇研究報告新發現的isoform 6。
偵測Isoform 6在各種癌症細胞株及星狀細胞瘤病人的表現量並進行分析，發現isoform 6只有在IMR32細胞株（神經母細胞瘤細胞株）中被偵測到；而在七個星狀細胞瘤的病人組織中僅有三個有表現，推論isoform 6可能主要存在於神經細胞中。
在免疫螢光染色的實驗中除了isoform 6之外，其他三種isoforms都可以和中心體聚集在一起，且在細胞中大量表現isoform 6會影響

Centrosome is the major microtubule organizing center in mammalian cells. The centrosome plays key roles in the formation of the mitotic spindle, cell polarity and cell locomotion. In a typical somatic cell, the centrosome is composed of a pair of centrioles that are surrounded by a mass of amorphous pericentriolar material(PCM).
The centrosomal-associated protein, hNinein, has been identified as a microtubules minus end capping, centrioles position, centrosome maturation and anchoring protein. Recently data reveal ninein CCII domain is associated with centrosomal targeting signal, regulating signal and phosphorylation sites, suggesting that this protein could contribute to centrosomal targeting signal as well as regulate asymmetry centrosomes.
In this report, we therefore examine whether four C-terminal of hNinein splicing isoforms, including isoform 1, isoform 2, isoform 5 and a newly finding isoform 6, represent differential characterization, expression and functional aspects.
To investigate the expression level of hNinein isoform 6 in variant cancer cell lines and astrocytoma patients. Isoform 6 only exist in IMR32 cancer cell line ( neuroblastoma cell line) and 42﹪astrocytoma patients. We speculate that isoform 6 might play a role and exist in nerve cell or neuron.
Using immunofluorescence assay, we show that all three hNinein isoforms are concentrated at centrosomes in HeLa cells, but not isoform 6. Overexpression of isoform 6 in HeLa cell might influence the distribution of

目錄


中文摘要………………………………………………………………….i
英文摘要…………………………………………………………………iii
英文縮寫表………………………………………………………………v
前言………………………………………………………………………1
材料與方法………………………………………………………………4
結果……………………………………………………………………..13
討論……………………………………………………………………..18
參考文獻………………………………………………………………..21
圖表……………………………………………………………………..24
附錄……………………………………………………………………..41

參考文獻
Bornens, M. Centrosome composition and microtubule anchoring mechanisms. Curr. Opin. Cell Biol. 14(1):25-34. 2002
Bouckson-castaing, V., Moudjou, M., Ferguson, D. J. P., Mucklow, S., Belkaid, Y., Milon, G., and Crocker, P. R. Microtubule characterization of ninein, a new coiled-coil protein of the centrosome. J Cell Sci. 109:179-190. 1996.
Brinkley, B. R. Microtubule organizing centers. Annu Rev Cell Dev Biol. 1:145-72. 1985.
Casenghi, M., Meraldi, P., Weinhart, U., Duncan, P. I., Korner, R., Nigg, E. A. Polo-like kinase 1 regulates Nlp, a centrosome protein involved in microtubule nucleation. Dev Cell. 5(1):113-25. 2003.
Chen, C. H., Howng, S. L., Cheng, T. S., Chou, M. H., Huang, C. Y., Hong, Y. R. Molecular characterization of human ninein protein: two distinct subdomains required for centrosomal targeting and regulating signals in cell cycle. Biochem Biophys Res Commun. 308(4):975-83. 2003.
Doxsey, S.J., Stein, P., Evans, L., Calarco, P.D., and Kirschner, M. Pericentrin, a highly conserved centrosome protein involved in microtubule organization. Cell. 76:639-650. 1994.
Fuller, S. D., Gowen, B. E., Reinsch, S., Sawyer, A., Buendia, B., Wepf, R., and Karsenti, E. The core of the mammalian centriole contains gamma-tubulin. Curr. Biol. 5(12):1384-1393. 1995.
Gergely, F., Karlsson, C., Still, I., Cowell, J., Kilmartin, J., Raff, J. W. The TACC domain identifies a family of centrosomal proteins that can interact with microtubules. Proc Natl Acad Sci . 97(26):14352-7. 2000.
Hong, Y. R., Chen, C. H., Chang, J. H., Wang, S., Sy, W. D., Chou, C. K., Howng, S. L. Cloning and characterization of a novel human ninein protein that interacts with the glycogen synthase kinase 3beta. Biochim Biophys Acta. 1492(2-3):513-6. 2000.
Hong, Y. R., Chen, C.H., Chuo, M. H., Liou, S. Y., Howng, S. L. Genomic organization and molecular characterization of the human ninein gene. Biochem Biophys Res Commun. 279(3):989-95. 2000.
Howng, S. L., Hsu, H. C., Cheng, T. S., Lee, Y. L., Chang, L. K., Lu, P. J., Hong, Y. R. A novel ninein-interaction protein, CGI-99, blocks ninein phosphorylation by GSK3