請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/27214
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 葉秀慧 | |
dc.contributor.author | Cho-Chun Lu | en |
dc.contributor.author | 盧卓群 | zh_TW |
dc.date.accessioned | 2021-06-12T17:58:08Z | - |
dc.date.available | 2018-12-31 | |
dc.date.copyright | 2008-02-20 | |
dc.date.issued | 2008 | |
dc.date.submitted | 2008-01-29 | |
dc.identifier.citation | [1] R. C. Lee, R. L. Feinbaum, and V. Ambros, 'The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14,' Cell, vol. 75, pp. 843-54, Dec 3 1993.
[2] Y. Lee, K. Jeon, J. T. Lee, S. Kim, and V. N. Kim, 'MicroRNA maturation: stepwise processing and subcellular localization,' EMBO J, vol. 21, pp. 4663-70, Sep 2 2002. [3] Y. Lee, M. Kim, J. Han, K. H. Yeom, S. Lee, S. H. Baek, and V. N. Kim, 'MicroRNA genes are transcribed by RNA polymerase II,' EMBO J, vol. 23, pp. 4051-60, Oct 13 2004. [4] X. Cai, C. H. Hagedorn, and B. R. Cullen, 'Human microRNAs are processed from capped, polyadenylated transcripts that can also function as mRNAs,' RNA, vol. 10, pp. 1957-66, Dec 2004. [5] A. M. Denli, B. B. Tops, R. H. Plasterk, R. F. Ketting, and G. J. Hannon, 'Processing of primary microRNAs by the Microprocessor complex,' Nature, vol. 432, pp. 231-5, Nov 11 2004. [6] R. I. Gregory, K. P. Yan, G. Amuthan, T. Chendrimada, B. Doratotaj, N. Cooch, and R. Shiekhattar, 'The Microprocessor complex mediates the genesis of microRNAs,' Nature, vol. 432, pp. 235-40, Nov 11 2004. [7] M. T. Bohnsack, K. Czaplinski, and D. Gorlich, 'Exportin 5 is a RanGTP-dependent dsRNA-binding protein that mediates nuclear export of pre-miRNAs,' RNA, vol. 10, pp. 185-91, Feb 2004. [8] E. C. Lai, 'Micro RNAs are complementary to 3' UTR sequence motifs that mediate negative post-transcriptional regulation,' Nat Genet, vol. 30, pp. 363-4, Apr 2002. [9] B. P. Lewis, C. B. Burge, and D. P. Bartel, 'Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets,' Cell, vol. 120, pp. 15-20, Jan 14 2005. [10] E. Berezikov, V. Guryev, J. van de Belt, E. Wienholds, R. H. Plasterk, and E. Cuppen, 'Phylogenetic shadowing and computational identification of human microRNA genes,' Cell, vol. 120, pp. 21-4, Jan 14 2005. [11] A. Rodriguez, S. Griffiths-Jones, J. L. Ashurst, and A. Bradley, 'Identification of mammalian microRNA host genes and transcription units,' Genome Res, vol. 14, pp. 1902-10, Oct 2004. [12] G. A. Calin, C. Sevignani, C. D. Dumitru, T. Hyslop, E. Noch, S. Yendamuri, M. Shimizu, S. Rattan, F. Bullrich, M. Negrini, and C. M. Croce, 'Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers,' Proc Natl Acad Sci U S A, vol. 101, pp. 2999-3004, Mar 2 2004. [13] M. J. Aukerman and H. Sakai, 'Regulation of flowering time and floral organ identity by a MicroRNA and its APETALA2-like target genes,' Plant Cell, vol. 15, pp. 2730-41, Nov 2003. [14] C. Z. Chen, L. Li, H. F. Lodish, and D. P. Bartel, 'MicroRNAs modulate hematopoietic lineage differentiation,' Science, vol. 303, pp. 83-6, Jan 2 2004. [15] V. Ambros, 'MicroRNA pathways in flies and worms: growth, death, fat, stress, and timing,' Cell, vol. 113, pp. 673-6, Jun 13 2003. [16] C. H. Lecellier, P. Dunoyer, K. Arar, J. Lehmann-Che, S. Eyquem, C. Himber, A. Saib, and O. Voinnet, 'A cellular microRNA mediates antiviral defense in human cells,' Science, vol. 308, pp. 557-60, Apr 22 2005. [17] E. Vigorito, K. L. Perks, C. Abreu-Goodger, S. Bunting, Z. Xiang, S. Kohlhaas, P. P. Das, E. A. Miska, A. Rodriguez, A. Bradley, K. G. Smith, C. Rada, A. J. Enright, K. M. Toellner, I. C. Maclennan, and M. Turner, 'microRNA-155 Regulates the Generation of Immunoglobulin Class-Switched Plasma Cells,' Immunity, vol. 27, pp. 847-59, Dec 2007. [18] R. Fiore and G. Schratt, 'MicroRNAs in synapse development: tiny molecules to remember,' Expert Opin Biol Ther, vol. 7, pp. 1823-31, Dec 2007. [19] G. A. Calin, C. D. Dumitru, M. Shimizu, R. Bichi, S. Zupo, E. Noch, H. Aldler, S. Rattan, M. Keating, K. Rai, L. Rassenti, T. Kipps, M. Negrini, F. Bullrich, and C. M. Croce, 'Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia,' Proc Natl Acad Sci U S A, vol. 99, pp. 15524-9, Nov 26 2002. [20] A. Cimmino, G. A. Calin, M. Fabbri, M. V. Iorio, M. Ferracin, M. Shimizu, S. E. Wojcik, R. I. Aqeilan, S. Zupo, M. Dono, L. Rassenti, H. Alder, S. Volinia, C. G. Liu, T. J. Kipps, M. Negrini, and C. M. Croce, 'miR-15 and miR-16 induce apoptosis by targeting BCL2,' Proc Natl Acad Sci U S A, vol. 102, pp. 13944-9, Sep 27 2005. [21] J. Takamizawa, H. Konishi, K. Yanagisawa, S. Tomida, H. Osada, H. Endoh, T. Harano, Y. Yatabe, M. Nagino, Y. Nimura, T. Mitsudomi, and T. Takahashi, 'Reduced expression of the let-7 microRNAs in human lung cancers in association with shortened postoperative survival,' Cancer Res, vol. 64, pp. 3753-6, Jun 1 2004. [22] S. M. Johnson, H. Grosshans, J. Shingara, M. Byrom, R. Jarvis, A. Cheng, E. Labourier, K. L. Reinert, D. Brown, and F. J. Slack, 'RAS is regulated by the let-7 microRNA family,' Cell, vol. 120, pp. 635-47, Mar 11 2005. [23] Y. Hayashita, H. Osada, Y. Tatematsu, H. Yamada, K. Yanagisawa, S. Tomida, Y. Yatabe, K. Kawahara, Y. Sekido, and T. Takahashi, 'A polycistronic microRNA cluster, miR-17-92, is overexpressed in human lung cancers and enhances cell proliferation,' Cancer Res, vol. 65, pp. 9628-32, Nov 1 2005. [24] S. L. Yu, H. Y. Chen, G. C. Chang, C. Y. Chen, H. W. Chen, S. Singh, C. L. Cheng, C. J. Yu, Y. C. Lee, H. S. Chen, T. J. Su, C. C. Chiang, H. N. Li, Q. S. Hong, H. Y. Su, C. C. Chen, W. J. Chen, C. C. Liu, W. K. Chan, K. C. Li, J. J. Chen, and P. C. Yang, 'MicroRNA Signature Predicts Survival and Relapse in Lung Cancer,' Cancer Cell, vol. 13, pp. 48-57, Jan 8 2008. [25] Y. Guo, Z. Chen, L. Zhang, F. Zhou, S. Shi, X. Feng, B. Li, X. Meng, X. Ma, M. Luo, K. Shao, N. Li, B. Qiu, K. Mitchelson, J. Cheng, and J. He, 'Distinctive microRNA profiles relating to patient survival in esophageal squamous cell carcinoma,' Cancer Res, vol. 68, pp. 26-33, Jan 1 2008. [26] A. S. Yu and E. B. Keeffe, 'Management of hepatocellular carcinoma,' Rev Gastroenterol Disord, vol. 3, pp. 8-24, Winter 2003. [27] Z. Y. Tang, 'Hepatocellular carcinoma--cause, treatment and metastasis,' World J Gastroenterol, vol. 7, pp. 445-54, Aug 2001. [28] C. M. Lee, S. N. Lu, C. S. Changchien, C. T. Yeh, T. T. Hsu, J. H. Tang, J. H. Wang, D. Y. Lin, C. L. Chen, and W. J. Chen, 'Age, gender, and local geographic variations of viral etiology of hepatocellular carcinoma in a hyperendemic area for hepatitis B virus infection,' Cancer, vol. 86, pp. 1143-50, Oct 1 1999. [29] M. W. Yu, Y. C. Yang, S. Y. Yang, S. W. Cheng, Y. F. Liaw, S. M. Lin, and C. J. Chen, 'Hormonal markers and hepatitis B virus-related hepatocellular carcinoma risk: a nested case-control study among men,' J Natl Cancer Inst, vol. 93, pp. 1644-51, Nov 7 2001. [30] M. W. Yu, S. W. Cheng, M. W. Lin, S. Y. Yang, Y. F. Liaw, H. C. Chang, T. J. Hsiao, S. M. Lin, S. D. Lee, P. J. Chen, C. J. Liu, and C. J. Chen, 'Androgen-receptor gene CAG repeats, plasma testosterone levels, and risk of hepatitis B-related hepatocellular carcinoma,' J Natl Cancer Inst, vol. 92, pp. 2023-8, Dec 20 2000. [31] C. L. Jopling, M. Yi, A. M. Lancaster, S. M. Lemon, and P. Sarnow, 'Modulation of hepatitis C virus RNA abundance by a liver-specific MicroRNA,' Science, vol. 309, pp. 1577-81, Sep 2 2005. [32] Y. Murakami, T. Yasuda, K. Saigo, T. Urashima, H. Toyoda, T. Okanoue, and K. Shimotohno, 'Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues,' Oncogene, vol. 25, pp. 2537-45, Apr 20 2006. [33] H. Kutay, S. Bai, J. Datta, T. Motiwala, I. Pogribny, W. Frankel, S. T. Jacob, and K. Ghoshal, 'Downregulation of miR-122 in the rodent and human hepatocellular carcinomas,' J Cell Biochem, vol. 99, pp. 671-8, Oct 15 2006. [34] L. Gramantieri, M. Ferracin, F. Fornari, A. Veronese, S. Sabbioni, C. G. Liu, G. A. Calin, C. Giovannini, E. Ferrazzi, G. L. Grazi, C. M. Croce, L. Bolondi, and M. Negrini, 'Cyclin G1 is a target of miR-122a, a microRNA frequently down-regulated in human hepatocellular carcinoma,' Cancer Res, vol. 67, pp. 6092-9, Jul 1 2007. [35] X. Zhang, L. He, Y. Lu, M. Liu, and X. Huang, 'Androgen receptor in primary hepatocellular carcinoma and its clinical significance,' Chin Med J (Engl), vol. 111, pp. 1083-6, Dec 1998. [36] Y. Takahashi, S. Kawate, M. Watanabe, J. Fukushima, S. Mori, and T. Fukusato, 'Amplification of c-myc and cyclin D1 genes in primary and metastatic carcinomas of the liver,' Pathol Int, vol. 57, pp. 437-42, Jul 2007. [37] X. Qin, H. Zhang, X. Zhou, C. Wang, X. Zhang, and L. Ye, 'Proliferation and migration mediated by Dkk-1/Wnt/beta-catenin cascade in a model of hepatocellular carcinoma cells,' Transl Res, vol. 150, pp. 281-94, Nov 2007. [38] D. F. Calvisi, S. Ladu, A. Gorden, M. Farina, E. A. Conner, J. S. Lee, V. M. Factor, and S. S. Thorgeirsson, 'Ubiquitous activation of Ras and Jak/Stat pathways in human HCC,' Gastroenterology, vol. 130, pp. 1117-28, Apr 2006. [39] O. Barad, E. Meiri, A. Avniel, R. Aharonov, A. Barzilai, I. Bentwich, U. Einav, S. Gilad, P. Hurban, Y. Karov, E. K. Lobenhofer, E. Sharon, Y. M. Shiboleth, M. Shtutman, Z. Bentwich, and P. Einat, 'MicroRNA expression detected by oligonucleotide microarrays: system establishment and expression profiling in human tissues,' Genome Res, vol. 14, pp. 2486-94, Dec 2004. [40] S. H. Yeh, M. W. Lin, S. F. Lu, D. C. Wu, S. F. Tsai, C. Y. Tsai, M. Y. Lai, H. C. Hsu, D. S. Chen, and P. J. Chen, 'Allelic loss of chromosome 4q21 approximately 23 associates with hepatitis B virus-related hepatocarcinogenesis and elevated alpha-fetoprotein,' Hepatology, vol. 40, pp. 847-54, Oct 2004. [41] W. L. Shih, M. W. Yu, P. J. Chen, S. H. Yeh, M. T. Lo, H. C. Chang, Y. F. Liaw, S. M. Lin, C. J. Liu, S. D. Lee, C. L. Lin, C. K. Hsiao, S. Y. Yang, and C. J. Chen, 'Localization of a susceptibility locus for hepatocellular carcinoma to chromosome 4q in a hepatitis B hyperendemic area,' Oncogene, vol. 25, pp. 3219-24, May 25 2006. [42] S. Galardi, N. Mercatelli, E. Giorda, S. Massalini, G. V. Frajese, S. A. Ciafre, and M. G. Farace, 'miR-221 and miR-222 expression affects the proliferation potential of human prostate carcinoma cell lines by targeting p27Kip1,' J Biol Chem, vol. 282, pp. 23716-24, Aug 10 2007. [43] C. le Sage, R. Nagel, D. A. Egan, M. Schrier, E. Mesman, A. Mangiola, C. Anile, G. Maira, N. Mercatelli, S. A. Ciafre, M. G. Farace, and R. Agami, 'Regulation of the p27(Kip1) tumor suppressor by miR-221 and miR-222 promotes cancer cell proliferation,' EMBO J, vol. 26, pp. 3699-708, Aug 8 2007. [44] R. Visone, L. Russo, P. Pallante, I. De Martino, A. Ferraro, V. Leone, E. Borbone, F. Petrocca, H. Alder, C. M. Croce, and A. Fusco, 'MicroRNAs (miR)-221 and miR-222, both overexpressed in human thyroid papillary carcinomas, regulate p27Kip1 protein levels and cell cycle,' Endocr Relat Cancer, vol. 14, pp. 791-8, Sep 2007. [45] N. Nagasue, H. Yukaya, Y. C. Chang, Y. Ogawa, H. Kohno, and A. Ito, 'Active uptake of testosterone by androgen receptors of hepatocellular carcinoma in humans,' Cancer, vol. 57, pp. 2162-7, Jun 1 1986. [46] N. Nagasue, H. Kohno, A. Yamanoi, T. Kimoto, Y. C. Chang, and T. Nakamura, 'Progesterone receptor in hepatocellular carcinoma. Correlation with androgen and estrogen receptors,' Cancer, vol. 67, pp. 2501-5, May 15 1991. [47] Y. Akao, Y. Nakagawa, and T. Naoe, 'let-7 microRNA functions as a potential growth suppressor in human colon cancer cells,' Biol Pharm Bull, vol. 29, pp. 903-6, May 2006. [48] K. Inamura, Y. Togashi, K. Nomura, H. Ninomiya, M. Hiramatsu, Y. Satoh, S. Okumura, K. Nakagawa, and Y. Ishikawa, 'let-7 microRNA expression is reduced in bronchioloalveolar carcinoma, a non-invasive carcinoma, and is not correlated with prognosis,' Lung Cancer, vol. 58, pp. 392-6, Dec 2007. [49] E. Bandres, E. Cubedo, X. Agirre, R. Malumbres, R. Zarate, N. Ramirez, A. Abajo, A. Navarro, I. Moreno, M. Monzo, and J. Garcia-Foncillas, 'Identification by Real-time PCR of 13 mature microRNAs differentially expressed in colorectal cancer and non-tumoral tissues,' Mol Cancer, vol. 5, p. 29, 2006. [50] M. V. Iorio, R. Visone, G. Di Leva, V. Donati, F. Petrocca, P. Casalini, C. Taccioli, S. Volinia, C. G. Liu, H. Alder, G. A. Calin, S. Menard, and C. M. Croce, 'MicroRNA signatures in human ovarian cancer,' Cancer Res, vol. 67, pp. 8699-707, Sep 15 2007. [51] O. Slaby, M. Svoboda, P. Fabian, T. Smerdova, D. Knoflickova, M. Bednarikova, R. Nenutil, and R. Vyzula, 'Altered Expression of miR-21, miR-31, miR-143 and miR-145 Is Related to Clinicopathologic Features of Colorectal Cancer,' Oncology, vol. 72, pp. 397-402, Jan 15 2008. [52] S. J. Greco and P. Rameshwar, 'MicroRNAs regulate synthesis of the neurotransmitter substance P in human mesenchymal stem cell-derived neuronal cells,' Proc Natl Acad Sci U S A, vol. 104, pp. 15484-9, Sep 25 2007. [53] Y. Chen and D. H. Gorski, 'Regulation of angiogenesis through a microRNA (miR-130a) that downregulates antiangiogenic homeobox genes GAX and HOXA5,' Blood, Oct 23 2007. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/27214 | - |
dc.description.abstract | MicroRNAs 是具有19至25個核苷酸的小片段RNA,它主要會辨認到 mRNA 的3端未轉譯區,形成不完全的配對而抑制基因的蛋白質表現。它在細胞裡扮演著很重要的功能,例如發育的調控,還有細胞的脂肪代謝、增生、分化及細胞的死亡和抗病毒機制等。另外,miRNA 的不正常表現可能和疾病的發生有相關,已有文獻指出 miRNA 在腫瘤細胞中可能扮演著致癌基因或是腫瘤抑制基因的角色,因此分析在腫瘤的形成過程中,有哪些的miRNA 有不正常的表現情形,可以幫助我們了解腫瘤的致病機轉。
我們根據 (1) 之前在文獻中利用microarray分析所指出在HCC中有顯著差異的miRNAs 或是可能有性別表現差異的 miRNAs;(2)經由網站預測出與肝癌形成可能相關基因之miRNAs;(3) 在肝臟細胞內有高度表現量的 miRNA;(4) 和位在第四對染色體上於肝癌好發易斷裂區的miRNAs,經由此種方式,我們共選擇了29 個miRNAs來進行本論文針對miRNA於HCC癌化過程中是否產生變化之分析。為探討miRNA之變化是否和病毒感染因子以及男女性別差異之特性相關,我們分析之臨床檢體主要包含了不同病毒感染因子相關之男女配對之tumor 及其non-tumor肝臟組織,並以配對 FNH (Focal nodular hyperplasia,肝臟結節性增生)肝臟組織當作對照組。我們使用定量real-time PCR 的方式去偵測肝組織內的 miRNA 的表現量,以t-test 分析並把p-value 小於0.05視為有顯著的差異。 在我們的研究中,我們發現有好幾個 miRNAs 在 HCC 內的表現量有明顯的改變。另外,我們也發現有些 miRNAs 在依病毒或是性別分群上有表現量的差異,但仍需要進一步證明。為了指出 miRNAs 在癌化過程中的作用機制,我們選擇了2個 miRNAs : mir-130a 和 mir-493-5p 做為進一步實驗。 因為我們發現mir-130a 和 mir-493-5p 在 HCC 的表現量是有顯著下降,且mir-130a 和 mir-493-5p是被預測會去調控 AR 的miRNAs,因此我們進行實驗想去探討mir-130a 或mir-493-5p 是否能辨認到 AR 並調控其表現。 我們所使用的方法是把具有AR的3端未轉譯區接在 pGL3-promoter 之後,轉染入人類肝癌細胞株,再使用感染 lentivirus 的方式去大量表現 mir-130a 或是 mir-493-5p。為了證實這辨識的特異性,我們把 mir-130a 和 mir-493-5p 所辨識的 seed site 做了4個核苷酸的突變。雖然在初步的實驗中,可以發現這兩個 miRNA 可以造成具有AR-3端未轉譯區的reporter 活性下降,但是這兩個 miRNAs 對於內生性 AR 的蛋白質表現量抑制的程度並不明顯。 結論,在我們的研究中指出有幾個miRNAs 在HCC 的表現量有顯著的差異,但它們在癌化過程中所扮演的角色,則需要更進一步的實驗去證明。 | zh_TW |
dc.description.abstract | MiRNAs are 19- to 25-nucleotide-long RNAs which can inhibit the protein synthesis by binding to the complementary sequences at the 3’-untranslated regions (3’-UTR) of target mRNAs. They play regulatory roles in various biological functions, including development, cell fate determination, proliferation, differentiation, cell death, and antiviral mechanisms etc. Intriguingly, an increasing list of miRNAs have been reported to function as putative oncogenes or tumor suppressor genes and showed aberrant expression patterns during carcinogenic process. Therefore, it is assumed that analysis of the miRNA expression patterns of caner cells can help researcher to understand the carcinogeneic mechanisms of specific tumors.
Aiming to study the functional involvement of miRNA in liver cancers, I started a pilot study by examining the expression patterns of numerous specific miRNAs in paired HCC and corresponding non-tumorous tissues from HBV-, HCV-, and nonB/nonC related HCCs collected both from the male and female patients. Such an approach can help to elucidate the correlation of specific miRNA expression pattern with the viral etiology and gender factor. The selected miRNAs in the current study were based on the criteria listed as followed. (1) The ones showing significant difference or possible gender difference revealed by the previous studies approached by microarray analysis; (2) the predicted miRNAs target genes which are associated with hepatocarcinogenesis; (3) the ones with high expression level in the liver tissues; and (4) the ones locating in the chromosome 4q region with frequent allelic loss. The assay used to determine the amount of miRNA in liver tissues is the quantitative real-time PCR analysis, which is assumed to be a much more reliable method for such analysis. The results from current analysis indeed pointed out several miRNAs showing significant different expression patterns between tumorous and non-tumorous tissues (p<0.05, t-test). Moreover, I have identified a few miRNAs showing viral etiological or gender difference, which still awaits further verification. To illustrate the functional mechanism of specific miRNA in carcinogenesis, I have chosen two miRNA, the mir-130a and mir-493-5, which were predicted to target to the androgen receptor (AR) gene, as an example in the subsequent functional analysis. Since these two miRNAs showed significant down-regulated expression in HCCs, I tried to test if these two miRNAs indeed target to AR gene and regulate its expression negatively. The strategy is to deliver the mi-RNA expression constructs into hepatoma cells with the luciferase reporter construct fused with the 3’–UTR of AR gene, which contains the putative recognition sites of both miRNAs. To verify the specificity of the recognition, I also included a reporter construct with mutations at the target recognition sequence as a control. Although the preliminary results showed down-regulation of reporter activity by both miRNAs, their effect on the AR protein expression is not evident. In conclusion, current study pointed out several miRNAs with significant changes in HCCs. Yet, their functional role in carcinogenic process warrants further clarification. | en |
dc.description.provenance | Made available in DSpace on 2021-06-12T17:58:08Z (GMT). No. of bitstreams: 1 ntu-97-R94445131-1.pdf: 8914616 bytes, checksum: b1cfb207c80b645738a792f2ecfdf380 (MD5) Previous issue date: 2008 | en |
dc.description.tableofcontents | 口試委員會審定書………………………………………………………………………i
誌謝…………………………………………………………………ii 中文摘要………………………………………………………………iii Abstract…………………………………………………………………v 第一章 序論……………………………………………………………………1 1-1.miRNA 的生合成及其調控基因的方式…………………………1 1-2. microRNA 所調控之正常生理功能………………………………2 1-3. microRNA與腫瘤的相關性………………………………………2 1-4.肝癌形成之原因與性別差異………………………………………4 1-5. microRNA與病毒感染的相關性…………………………………4 1-6.肝癌的microRNA的研究……………………………………………5 第二節 研究假說、目的、與研究策略 2-1: 研究假說與目的…………………………………………………6 2-2: 先前研究的限制…………………………………………………6 2-3: 研究策略…………………………………………………………7 第二章 實驗材料與方法………………………………………………8 1.臨床肝臟組織檢體的選擇及來源…………………………………8 2.Total RNA 的萃取…………………………………………………8 3. 29個 miRNAs 的選擇………………………………………………9 4. 定量 Real-time PCR 的分析……………………………………11 5. 數據的分析………………………………………………………12 6. 細胞培養…………………………………………………………13 7. 質體及其建構…………………………………………………13 8. 質體轉型於E. coil……………………………………………17 9. Lentivirus 的製備及感染細胞…………………………………17 10. 冷光酶測定分析…………………………………………………18 11. LNCapP cell 的感染處理………………………………………19 12. 蛋白質處理及定量………………………………………………20 13. SDS-PAGE 蛋白質膠體電泳………………………………………20 14. 核醣核酸保護分析試驗…………………………………………21 第三章 結果……………………………………………………………23 I-1. 29個 miRNAs 的選擇……………………………………………23 I-2. HCC組織 non-tumor 和 tumor 的 miRNA 整體表現量分析…23 I-3. miRNA在配對 HCC 與其 non-tumor組織中癌化過程中表現量的改變分析……………………………………………24 I-4. miRNAs 和 virus etiology 的相關性分析…………………25 I-5. miRNAs 和性別的相關性分析………………………………25 I-6. miRNAs 和病毒及性別的相關性分析………………………26 II.探討男性荷爾蒙受體 (androgen receptor) 是否為mir-130a 和 mir-493-5p 的標的基因…………………………………………27 II-1.實驗的假設…………………………………………………27 II-2.實驗設計……………………………………………………28 第四章 討論……………………………………………………………31 第五章 參考文獻……………………………………………………37 第六章 圖表…………………………………………………………45 | |
dc.language.iso | zh-TW | |
dc.title | MicroRNA在肝癌形成過程中表現量的改變及相關致癌機轉之研究 | zh_TW |
dc.title | The Change of MicroRNA Expression Patterns and A Related Carcinogenic Mechanism in Hepatocarcinogenesis | en |
dc.type | Thesis | |
dc.date.schoolyear | 96-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳培哲,王弘毅 | |
dc.subject.keyword | 肝癌,男性荷爾蒙受體, | zh_TW |
dc.subject.keyword | microRNA,HCC,Hepatocarcinogenesis,androgen receptor, | en |
dc.relation.page | 61 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2008-01-30 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 微生物學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-97-1.pdf 目前未授權公開取用 | 8.71 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。