Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/30737
標題: 利用昆蟲桿狀病毒表現載體研究哺乳動物細胞株之基因轉導
The Use of Baculovirus Expression Vector for Studying The Gene Transduction of Mammalian Cells
作者: Jessica, C.Y.Lu
呂家芸
關鍵字: http://etds.lib.nchu.edu.tw/etdservice/view_metadata?etdun=U0005-1407200911203800
加州苜蓿夜蛾核多角體病毒
重組桿狀病毒
哺乳動物細胞株
顆粒蛋白前體
癌細胞
炎症
出版社: 昆蟲學系所
引用: Adair, R., Patel, A.H., Corless, L., Griffin, S., Rowlands, D.J. and McCormick, C.J., (2009). ''Expression of hepatitis C virus (HCV) structural proteins in trans facilitates encapsidation and transmission of HCV subgenomic RNA''. J Gen Virol, 90 (Pt 4):833-842. Bai, B., Hu, Q., Hu, H., Zhou, P., Shi, Z., Meng, J., Lu, B., Huang, Y., Mao, P. and Wang, H., (2008). ''Virus-like particles of SARS-like coronavirus formed by membrane proteins from different origins demonstrate stimulating activity in human dendritic cells''. PLoS ONE, 3 (7):e2685. Belyaev, A.S. and Roy, P., (1993). ''Development of baculovirus triple and quadruple expression vectors: co-expression of three or four bluetongue virus proteins and the synthesis of bluetongue virus-like particles in insect cells''. Nucleic Acids Res, 21 (5):1219-1223. Carlson, D.L., (2009). ''Necrotizing sialometaplasia: a practical approach to the diagnosis''. Arch Pathol Lab Med, 133 (5):692-698. Chen, J., Somanath, P.R., Razorenova, O., Chen, W.S., Hay, N., Bornstein, P. and Byzova, T.V., (2005). ''Akt1 regulates pathological angiogenesis, vascular maturation and permeability in vivo''. Nat Med, 11 (11):1188-1196. Chen, J.S., Liu, J.C., Shen, L., Rau, K.M., Kuo, H.P., Li, Y.M., Shi, D., Lee, Y.C., Chang, K.J. and Hung, M.C., (2004). ''Cancer-specific activation of the survivin promoter and its potential use in gene therapy''. Cancer Gene Ther, 11 (11):740-747. Chen, S.F., Lin, C.Y., Chang, Y.C., Li, J.W., Fu, E., Chang, F.N., Lin, Y.L. and Nieh, S., (2009). ''Effects of small interfering RNAs targeting Fascin on gene expression in oral cancer cells''. J Oral Pathol Med. Chuang, C.K., Sung, L.Y., Hwang, S.M., Lo, W.H., Chen, H.C. and Hu, Y.C., (2007). ''Baculovirus as a new gene delivery vector for stem cell engineering and bone tissue engineering''. Gene Ther, 14 (19):1417-1424. Chuang, C.K., Wong, T.H., Hwang, S.M., Chang, Y.H., Chen, G.Y., Chiu, Y.C., Huang, S.F. and Hu, Y.C., (2009). ''Baculovirus transduction of mesenchymal stem cells: in vitro responses and in vivo immune responses after cell transplantation''. Mol Ther, 17 (5):889-896. Condreay, J.P. and Kost, T.A., (2007). ''Baculovirus expression vectors for insect and mammalian cells''. Curr Drug Targets, 8 (10):1126-1131. Dan, H.C., Cooper, M.J., Cogswell, P.C., Duncan, J.A., Ting, J.P. and Baldwin, A.S., (2008). ''Akt-dependent regulation of NF-{kappa}B is controlled by mTOR and Raptor in association with IKK''. Genes Dev, 22 (11):1490-1500. Daniel, R., Daniels, E., He, Z. and Bateman, A., (2003). ''Progranulin (acrogranin/PC cell-derived growth factor/granulin-epithelin precursor) is expressed in the placenta, epidermis, microvasculature, and brain during murine development''. Dev Dyn, 227 (4):593-599. Daniel, R., He, Z., Carmichael, K.P., Halper, J. and Bateman, A., (2000). ''Cellular localization of gene expression for progranulin''. J Histochem Cytochem, 48 (7):999-1009. Desai, A., Konda, V.R., Darland, G., Austin, M., Prabhu, K.S., Bland, J.S., Carroll, B.J. and Tripp, M.L., (2009). ''META060 inhibits multiple kinases in the NF-kappaB pathway and suppresses LPS--mediated inflammation in vitro and ex vivo''. Inflamm Res, 58 (5):229-234. Donald, C.D., Laddu, A., Chandham, P., Lim, S.D., Cohen, C., Amin, M., Gerton, G.L., Marshall, F.F. and Petros, J.A., (2001). ''Expression of progranulin and the epithelin/granulin precursor acrogranin correlates with neoplastic state in renal epithelium''. Anticancer Res, 21 (6A):3739-3742. Dunne, A. and O''Neill, L.A., (2003). ''The interleukin-1 receptor/Toll-like receptor superfamily: signal transduction during inflammation and host defense''. Sci STKE, 2003 (171):re3. Eriksen, J.L. and Mackenzie, I.R., (2008). ''Progranulin: normal function and role in neurodegeneration''. J Neurochem, 104 (2):287-297. Ernst, W., Schinko, T., Spenger, A., Oker-Blom, C. and Grabherr, R., (2006). ''Improving baculovirus transduction of mammalian cells by surface display of a RGD-motif''. J Biotechnol, 126 (2):237-240. Escobar, G.A., McIntyre, R.C., Jr., Moore, E.E., Gamboni-Robertson, F. and Banerjee, A., (2006). ''Clathrin heavy chain is required for TNF-induced inflammatory signaling''. Surgery, 140 (2):268-272. Faissner, A., Heck, N., Dobbertin, A. and Garwood, J., (2006). ''DSD-1-Proteoglycan/Phosphacan and receptor protein tyrosine phosphatase-beta isoforms during development and regeneration of neural tissues''. Adv Exp Med Biol, 557:25-53. Fipaldini, C., Bellei, B. and La Monica, N., (1999). ''Expression of hepatitis C virus cDNA in human hepatoma cell line mediated by a hybrid baculovirus-HCV vector''. Virology, 255 (2):302-311. Frebel, K. and Wiese, S., (2006). ''Signalling molecules essential for neuronal survival and differentiation''. Biochem Soc Trans, 34 (Pt 6):1287-1290. Garofalo, R.S., Orena, S.J., Rafidi, K., Torchia, A.J., Stock, J.L., Hildebrandt, A.L., Coskran, T., Black, S.C., Brees, D.J., Wicks, J.R., McNeish, J.D. and Coleman, K.G., (2003). ''Severe diabetes, age-dependent loss of adipose tissue, and mild growth deficiency in mice lacking Akt2/PKB beta''. J Clin Invest, 112 (2):197-208. He, Y., Li, J., Heck, S., Lustigman, S. and Jiang, S., (2006). ''Antigenic and immunogenic characterization of recombinant baculovirus-expressed severe acute respiratory syndrome coronavirus spike protein: implication for vaccine design''. J Virol, 80 (12):5757-5767. He, Z. and Bateman, A., (2003). ''Progranulin (granulin-epithelin precursor, PC-cell-derived growth factor, acrogranin) mediates tissue repair and tumorigenesis''. J Mol Med, 81 (10):600-612. He, Z., Ong, C.H., Halper, J. and Bateman, A., (2003). ''Progranulin is a mediator of the wound response''. Nat Med, 9 (2):225-229. Huang, T.T. and Miyamoto, S., (2001). ''Postrepression activation of NF-kappaB requires the amino-terminal nuclear export signal specific to IkappaBalpha''. Mol Cell Biol, 21 (14):4737-4747. Jin, R., Lv, Z., Chen, Q., Quan, Y., Zhang, H., Li, S., Chen, G., Zheng, Q., Jin, L., Wu, X., Chen, J. and Zhang, Y., (2008). ''Safety and immunogenicity of H5N1 influenza vaccine based on baculovirus surface display system of Bombyx mori''. PLoS ONE, 3 (12):e3933. Jones, M.B., Michener, C.M., Blanchette, J.O., Kuznetsov, V.A., Raffeld, M., Serrero, G., Emmert-Buck, M.R., Petricoin, E.F., Krizman, D.B., Liotta, L.A. and Kohn, E.C., (2003). ''The granulin-epithelin precursor/PC-cell-derived growth factor is a growth factor for epithelial ovarian cancer''. Clin Cancer Res, 9 (1):44-51. Kandel, E.S., Skeen, J., Majewski, N., Di Cristofano, A., Pandolfi, P.P., Feliciano, C.S., Gartel, A. and Hay, N., (2002). ''Activation of Akt/protein kinase B overcomes a G(2)/m cell cycle checkpoint induced by DNA damage''. Mol Cell Biol, 22 (22):7831-7841. Kane, L.P., Shapiro, V.S., Stokoe, D. and Weiss, A., (1999). ''Induction of NF-kappaB by the Akt/PKB kinase''. Curr Biol, 9 (11):601-604. Kaneko, H., Suzuki, H., Abe, T., Miyano-Kurosaki, N. and Takaku, H., (2006). ''Inhibition of HIV-1 replication by vesicular stomatitis virus envelope glycoprotein pseudotyped baculovirus vector-transduced ribozyme in mammalian cells''. Biochem Biophys Res Commun, 349 (4):1220-1227. Karin, M. and Ben-Neriah, Y., (2000). ''Phosphorylation meets ubiquitination: the control of NF-[kappa]B activity''. Annu Rev Immunol, 18:621-663. Kinnunen, K., Kalesnykas, G., Mahonen, A.J., Laidinen, S., Holma, L., Heikura, T., Airenne, K., Uusitalo, H. and Yla-Herttuala, S., (2009). ''Baculovirus is an efficient vector for the transduction of the eye: comparison of baculovirus- and adenovirus-mediated intravitreal vascular endothelial growth factor D gene transfer in the rabbit eye''. J Gene Med, 11 (5):382-389. Kojima, Y., Ono, K., Inoue, K., Takagi, Y., Kikuta, K.I., Nishimura, M., Yoshida, Y., Nakashima, Y., Matsumae, H., Furukawa, Y., Mikuni, N., Nobuyoshi, M., Kimura, T., Kita, T. and Tanaka, M., (2009). ''Progranulin expression in advanced human atherosclerotic plaque''. Atherosclerosis.(in press) Kost, T.A., Condreay, J.P. and Jarvis, D.L., (2005). ''Baculovirus as versatile vectors for protein expression in insect and mammalian cells''. Nat Biotechnol, 23 (5):567-575. Lee, C.K., Lee, E.Y., Kim, Y.G., Mun, S.H., Moon, H.B. and Yoo, B., (2008). ''Alpha-lipoic acid inhibits TNF-alpha induced NF-kappa B activation through blocking of MEKK1-MKK4-IKK signaling cascades''. Int Immunopharmacol, 8 (2):362-370. Liao, C.T., Chang, J.T., Wang, H.M., Ng, S.H., Hsueh, C., Lee, L.Y., Lin, C.H., Chen, I.H., Huang, S.F., Cheng, A.J. and Yen, T.C., (2007). ''Survival in squamous cell carcinoma of the oral cavity: differences between pT4 N0 and other stage IVA categories''. Cancer, 110 (3):564-571. Liau, L.M., Lallone, R.L., Seitz, R.S., Buznikov, A., Gregg, J.P., Kornblum, H.I., Nelson, S.F. and Bronstein, J.M., (2000). ''Identification of a human glioma-associated growth factor gene, granulin, using differential immuno-absorption''. Cancer Res, 60 (5):1353-1360. Lu, R. and Serrero, G., (2000). ''Inhibition of PC cell-derived growth factor (PCDGF, epithelin/granulin precursor) expression by antisense PCDGF cDNA transfection inhibits tumorigenicity of the human breast carcinoma cell line MDA-MB-468''. Proc Natl Acad Sci U S A, 97 (8):3993-3998. Marfella, R., Cacciapuoti, F., Grassia, A., Manfredi, E., De Maio, G., Caruso, G., Pepe, M. and Nittolo, G., (2006). ''Role of the ubiquitin-proteasome system in carotid plaque instability in diabetic patients''. Acta Cardiol, 61 (6):630-636. Matilainen, H., Makela, A.R., Riikonen, R., Saloniemi, T., Korhonen, E., Hyypia, T., Heino, J., Grabherr, R. and Oker-Blom, C., (2006). ''RGD motifs on the surface of baculovirus enhance transduction of human lung carcinoma cells''. J Biotechnol, 125 (1):114-126. Matsuo, E., Tani, H., Lim, C., Komoda, Y., Okamoto, T., Miyamoto, H., Moriishi, K., Yagi, S., Patel, A.H., Miyamura, T. and Matsuura, Y., (2006). ''Characterization of HCV-like particles produced in a human hepatoma cell line by a recombinant baculovirus''. Biochem Biophys Res Commun, 340 (1):200-208. Monami, G., Gonzalez, E.M., Hellman, M., Gomella, L.G., Baffa, R., Iozzo, R.V. and Morrione, A., (2006). ''Proepithelin promotes migration and invasion of 5637 bladder cancer cells through the activation of ERK1/2 and the formation of a paxillin/FAK/ERK complex''. Cancer Res, 66 (14):7103-7110. Naumann, M., (2007). ''Control of the NF-kappaB inhibitor IkappaBalpha in pathogen infection''. Biochem Soc Trans, 35 (Pt 2):267-269. Novotny, N.M., Markel, T.A., Crisostomo, P.R. and Meldrum, D.R., (2008). ''Differential IL-6 and VEGF secretion in adult and neonatal mesenchymal stem cells: role of NFkB''. Cytokine, 43 (2):215-219. Ong, C.H. and Bateman, A., (2003). ''Progranulin (granulin-epithelin precursor, PC-cell derived growth factor, acrogranin) in proliferation and tumorigenesis''. Histol Histopathol, 18 (4):1275-1288. Prel, A., Le Gall-Recule, G., Cherbonnel, M., Grasland, B., Amelot, M. and Jestin, V., (2007). ''Assessment of the protection afforded by triple baculovirus recombinant coexpressing H5, N3, M1 proteins against a homologous H5N3 low-pathogenicity avian influenza virus challenge in Muscovy ducks''. Avian Dis, 51 (1 Suppl):484-489. Ramaswamy, S., Nakamura, N., Vazquez, F., Batt, D.B., Perera, S., Roberts, T.M. and Sellers, W.R., (1999). ''Regulation of G1 progression by the PTEN tumor suppressor protein is linked to inhibition of the phosphatidylinositol 3-kinase/Akt pathway''. Proc Natl Acad Sci U S A, 96 (5):2110-2115. Safdar, A. and Cox, M.M., (2007). ''Baculovirus-expressed influenza vaccine. A novel technology for safe and expeditious vaccine production for human use''. Expert Opin Investig Drugs, 16 (7):927-934. Seo, N.S., Hollister, J.R. and Jarvis, D.L., (2001). ''Mammalian glycosyltransferase expression allows sialoglycoprotein production by baculovirus-infected insect cells''. Protein Expr Purif, 22 (2):234-241. Shi, X. and Jarvis, D.L., (2007). ''Protein N-glycosylation in the baculovirus-insect cell system''. Curr Drug Targets, 8 (10):1116-1125. Simpkins, F.A., Devoogdt, N.M., Rasool, N., Tchabo, N.E., Alejandro, E.U., Kamrava, M.M. and Kohn, E.C., (2008). ''The alarm anti-protease, secretory leukocyte protease inhibitor, is a proliferation and survival factor for ovarian cancer cells''. Carcinogenesis, 29 (3):466-472. Smith, G.E., Summers, M.D. and Fraser, M.J., (1983). ''Production of human beta interferon in insect cells infected with a baculovirus expression vector''. Mol Cell Biol, 3 (12):2156-2165. Song, G., Ouyang, G. and Bao, S., (2005). ''The activation of Akt/PKB signaling pathway and cell survival''. J Cell Mol Med, 9 (1):59-71. Song, Y.S., Narasimhan, P., Kim, G.S., Jung, J.E., Park, E.H. and Chan, P.H., (2008). ''The role of Akt signaling in oxidative stress mediates NF-kappaB activation in mild transient focal cerebral ischemia''. J Cereb Blood Flow Metab, 28 (12):1917-1926. Staal, S.P., Hartley, J.W. and Rowe, W.P., (1977). ''Isolation of transforming murine leukemia viruses from mice with a high incidence of spontaneous lymphoma''. Proc Natl Acad Sci U S A, 74 (7):3065-3067. Sudheerkumar, P., Shiras, A., Das, G., Jagtap, J.C., Prasad, V. and Shastry, P., (2008). ''Independent activation of Akt and NF-kappaB pathways and their role in resistance to TNF-alpha mediated cytotoxicity in gliomas''. Mol Carcinog, 47 (2):126-136. Tolkatchev, D., Malik, S., Vinogradova, A., Wang, P., Chen, Z., Xu, P., Bennett, H.P., Bateman, A. and Ni, F., (2008). ''Structure dissection of human progranulin identifies well-folded granulin/epithelin modules with unique functional activities''. Protein Sci, 17 (4):711-724. Tu, M.L., Wang, H.Q., Chen, L.J., Lu, J.C., Jiang, F., Liang, J.H., Xu, D.G. and Li, D.S., (2009). ''Involvement of Akt1/protein kinase Balpha in tumor conditioned medium-induced endothelial cell migration and survival in vitro''. J Cancer Res Clin Oncol.(in press) van Oers, M.M. and Vlak, J.M., (2007). ''Baculovirus genomics''. Curr Drug Targets, 8 (10):1051-1068. Vanarsdall, A.L., Mikhailov, V.S. and Rohrmann, G.F., (2007). ''Baculovirus DNA replication and processing''. Curr Drug Targets, 8 (10):1096-1102. Wang, W., Hayashi, J., Kim, W.E. and Serrero, G., (2003). ''PC cell-derived growth factor (granulin precursor) expression and action in human multiple myeloma''. Clin Cancer Res, 9 (6):2221-2228. Wu, Q., Fang, L., Wu, X., Li, B., Luo, R., Yu, Z., Jin, M., Chen, H. and Xiao, S., (2009). ''A pseudotype baculovirus-mediated vaccine confers protective immunity against lethal challenge with H5N1 avian influenza virus in mice and chickens''. Mol Immunol. Wullaert, A., Heyninck, K., Janssens, S. and Beyaert, R., (2006). ''Ubiquitin: tool and target for intracellular NF-kappaB inhibitors''. Trends Immunol, 27 (11):533-540. Yang, Z.Z., Tschopp, O., Baudry, A., Dummler, B., Hynx, D. and Hemmings, B.A., (2004). ''Physiological functions of protein kinase B/Akt''. Biochem Soc Trans, 32 (Pt 2):350-354. Zaidi, G. and Supakar, P.C., (2003). ''Identification of a nuclear protein interacting with a novel site on rat androgen receptor promoter after transcription factor NFkB is displaced from adjacent site''. Mol Biol Rep, 30 (2):121-125. Zhou, J. and Blissard, G.W., (2008). ''Display of heterologous proteins on gp64null baculovirus virions and enhanced budding mediated by a vesicular stomatitis virus G-stem construct''. J Virol, 82 (3):1368-1377. Zhu, J., Nathan, C., Jin, W., Sim, D., Ashcroft, G.S., Wahl, S.M., Lacomis, L., Erdjument-Bromage, H., Tempst, P., Wright, C.D. and Ding, A., (2002). ''Conversion of proepithelin to epithelins: roles of SLPI and elastase in host defense and wound repair''. Cell, 111 (6):867-878.
摘要: The present study used baculovirus expression vector system that is based on the budded form of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) to construct recombinant viruses, vAtEsL, carrying three promoters, i.e., CMVie-enhance promoter, p10 promoter, and T7 promoter, to drive the enhanced green fluorescent protein (EGFP) as a reporter and another recombinant virus carrying a survivin promoter to drive luciferase. There were different transduction rates among 19 mammalian cell lines. The bone cancer cell line (U2-osl) had the highest transduction rate followed by normal oral cell line (NHOK) and oral cancer cell lines (OECM-1, SAS and OC3). This finding could possibly be applied to study of oral cancers or to be used as its early diagnostic tool. The survivin promoter expressed successfully in the plasmid (pActEsB) to drive bax gene and triggered apoptosis; whereas its constructed recombinant baculovirus, vActEsB, was incapable of expressing it being due to low transduction rate, gene homology and/or variation in construction strategy. The other recombinant baculovirus, viz., vFuPiEG, was constructed based on FasBacI vector containing ubiquitin promoter to drive progranulin (PGRN) gene and internal ribosome entry site (IRES) to express EGFP. Since PGRN was expressed at a sufficient amount in H293 cells endogenously, its over-expression could not help to study the relationship between PGRN and NF-κB in inflammation. Therefore, this study underwent siRNA treatment to knock down PGRN expression in H293 cells, resulting in decrease of NF-κB activity. In addition, treatment of C6 malignant glioma cells and normal glial cells from rat with PGRN siRNA exhibited effect of PGRN on Akt activity. The results showed that PGRN knockdown was able to increase total Akt expression but to decrease phosphorylated Akt in the normal glial cells. Conversely, this knockdown did not obviously influence the glioma cells. Based upon the preliminary results in this study, it seems that the baculovirus expression vector system could potentially be employed in investigations on human cancer diseases and inflammation.
本文利用加州苜蓿夜蛾核多角體病毒(Autographa californica nuclopolyhedrovirus, AcMNPV )表現載體系統,構築含CMVie-enhance promoter、p10 promoter及T7 promoter等三個promoters驅動水母螢光基因(EGFP)及survivin promoter驅動Luciferase基因之重組桿狀病毒(vAtEsL),在感染19種之哺乳動物細胞株中,發現其在不同哺乳動物細胞之轉染力不一,以骨癌細胞(U2-osl)的轉染力最佳,口腔正常細胞(NHOK)與口腔癌細胞(OECM-1、SAS及OC3)次之,此對口腔癌可以應用於研究其致病機制或早期診斷之工具。另外,構築以survivin promoter驅動bax基因之重組桿狀病毒,結果其可藉由重組桿狀病毒的質體(pActEsB)在癌細胞株表現,且引起細胞凋亡,惟組成桿狀病毒(vActEsB)後,卻因轉染力不穩定或太低,以及在重組期間homology問題,質體構築設計之差異等因素,造成bax基因在轉染後並無理想的表達。本文的下半部份集中於探討醣基化蛋白 PGRN與哺乳動物內訊息傳遞之關係,雖先以FasBacI vector為基礎所構築的重組桿狀病毒(vFuPiEG),以ubiquitin promoter驅動醣基化蛋白 PGRN,並用IRES 啟動EGFP的報導基因,但由於試驗過程中發現H293細胞內的PGRN本底表達已經足夠,重組桿狀病毒所生產的PGRN無助於對該蛋白的功能性探討。因此,本試驗進行siRNA處理,對H293細胞之本底PGRN進行knockdown後,則發現NF-κB 的活性會隨PGRN之降低而下降。另外,以PGRN siRNA處理老鼠腦膠質瘤細胞(glioma cell,C6)與正常膠質細胞(glial cell),初探PGRN knock down後在Akt pathway之反應,試驗結果發現在正常的膠質細胞,經PGRN knockdown後Akt表達會上升,而磷酸化的AKT則降低;但在腫瘤細胞上, PGRN knockdown後對Akt通路無明顯影響。本文之初步結果顯示昆蟲桿狀病毒表現載體,在人類癌症及炎症之研究上似具有應用之潛力。
URI: http://hdl.handle.net/11455/30737
其他識別: U0005-1407200911203800
Appears in Collections:昆蟲學系

文件中的檔案:

取得全文請前往華藝線上圖書館

Show full item record
 
TAIR Related Article
 
Citations:


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.