臺灣博碩士論文加值系統

RAF-MEK-ERK是控制細胞增殖、分化和存亡的重要訊息傳遞路徑。最近的報導指出，B-RAF的突變在人類癌症中有很高的發生率，
B-RAF發生突變的exon 11和 exon 15位置分別在glycine-rich loop和活化作用區。在B-RAF突變中， V600E是在人類癌症中最常發現的突變位點，包括黑色素細胞癌及甲狀腺癌。在此次的論文研究，我們取得86例的癌症，其中包括62例的肺癌與24例的食道癌，進行聚合酶連鎖反應（PCR）放大BRAF exon 11和15以及對DNA序列進行突變之分析。在分析的結果中我們並沒有在這兩個exon中發現任何的突變，這顯示BRAF的突變在肺癌及食道癌中可能較少，而這樣的結論還需要更多的樣品來分析證實。

The RAF-MEK-ERK is an important signaling pathway that controls cellular proliferation, differentiation and survival. Recent reports indicate that R-RAF is mutated at a high frequency in human cancer. The mutations are clustered in the glycine-rich loop and activation segment which are encoded by exon 11 and exon 15, respectively. Among these mutations, V600E is the most prevalent found in varieties of human cancers, include melanoma and thyroid carcinomas. In this thesis, we analyzed 86 human cancer specimens, including 62 lung cancers and 24 esophageal cancers, for the mutation of exons 11 and 15 by PCR and direct DNA sequencing. However, we can not detect any mutation in these two exons in these clinical samples, these results suggest indicating that BRAF mutation might be rare and analysis of larger sample size is needed to confirmed this conclusion.

目 錄
中文摘要 ………………………………………………………………1
英文摘要 ………………………………………………………………2
背景介紹 ………………………………………………………………3
材料……………………………………………………………………15
方法……………………………………………………………………17
結果……………………………………………………………………19
討論……………………………………………………………………21
參考資料………………………………………………………………23
圖表……………………………………………………………………27

Barbacid, M. ras genes. Annu Rev Biochem 56: 779-827 (1987).
Brose, M. S., Volpe, P., Feldman, M., Kumar, M., Rishi, I., Gerrero, R., Einhorn, E., Herlyn, M., Minna, J., Nicholson, A., Roth, J. A., Albelda, S. M., Davies, H., Cox, C., Brignell, G., Stephens, P., Futreal, P. A., Wooster, R., Stratton, M. R. & Weber, B. L. BRAF and RAS mutations in human lung cancer and melanoma. Cancer Res 62: 6997-7000 (2002).
Carney, D. N. Lung Cancer Biology. Semin Radiat Oncol 5: 4-10 (1995).
Chen, C. L., Chi, C. W., Chang, K. W. & Liu, T. Y. Safrole-like DNA adducts in oral tissue from oral cancer patients with a betel quid chewing history. Carcinogenesis 20: 2331-2334 (1999)
Daum, G., Eisenmann-Tappe, I., Fries, H. W., Troppmair, J. & Rapp, U. R. The ins and outs of Raf kinases. Trends Biochem Sci 19: 474-480 (1994).
Davies, H., Bignell, G. R., Cox, C., Stephens, P., Edkins, S., Clegg, S., Teague, J., Woffendin, H., Garnett, M. J. & Bottomley, W. Mutations of the BRAF gene in human cancer. Nature 417: 949-954 (2002).
Eychene, A., Barnier, J. V., Apiou, F., Dutrillaux, B. & Calothy, G. Chromosomal assignment of two human B-raf(Rmil) proto-oncogene loci: B-raf-1 encoding the p94Braf/Rmil and B-raf-2, a processed pseudogene. Oncogene 7: 1657-1660 (1992).
Flood, W. A. & Forastiere, A. A. Esophageal cancer. Curr Opin Oncol 7: 381-386 (1995).
Fransen, K., Klintenas, M., Osterstrom, A., Dimberg, J., Monstein, H. J. & Soderkvist, P. Mutation analysis of the BRAF, ARAF and RAF-1 genes in human colorectal adenocarcinomas. Carcinogenesis 25: 527-533 (2004).
Garnett, M. J. & Marais, R. Guilty as charged: B-RAF is a human oncogene. Cancer Cell 6: 313-319 (2004).
Houben, R. Becker, J. C., Kappel, A., Terheyden, P., Brocker, E. B., Goetz, R. & Rapp, U. R. Constitutive activation of the Ras-Raf signaling