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| Inhibition of Proto-oncogene Pokemon Expression by siRNA Expression Vector in SW480 Cells |
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Abstract Abstract To investigate the inhibitory effects of siRNA expression vector on proto-oncogene Pokemon expression in SW480 cells and to provide experimental basis for further research about the biological function of Pokemon. siRNA expression vectors were constructed to express a short hairpin RNA to Pokemon. The recombinants were transfected into SW480 cells with liposome. Cellular morphology and transfection efficiency were observed. The expression of Pokemon were checked by real-time fluorescence quantitative PCR and Western blot. The transfection efficiency was about 36% and the cellular morphology changed greatly at 24h after transfection. siRNA expression vectors could specific reduce the expression of Pokemon mRNA and protein in SW480 cells. Compared with negative control, the inhibition ratio of Pokemon mRNA expression was 67.7% and the inhibition ratio of Pokemon protein was 73.6% respectively. siRNA expression vectors were successfully established that could effectively inhibit the expression of Pokemon in SW480 cells.
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Received: 19 March 2009
Published: 02 July 2009
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| ［1］ Maeda T, Hobbs R M, Merghoub T, et al. Role of the protooncogene Pokemon in cellular transformation and ARF repression. Nature, 2005, 433 (7023): 278~285 ［2］ Maeda T, Hobbs R M, Pandolfi PP. The transcription factor Pokemon: a new key player in cancer pathogenesis. Cancer Res, 2005, 65 (19): 8575~8578 ［3］ 蔡思, 于永春. 特殊的原癌基因Pokemon. 国际外科学杂志, 2006, 33 (6): 464~467 Cai S, Yu Y C. International Journal of Surgery, 2006, 33 (6): 464~467 ［4］ Vorhies J S, Nemunaitis J J. Synthetic vs. natural/biodegradable polymers for delivery of shRNAbased cancer therapies. Methods Mol Biol, 2009, 480: 11~29 ［5］ Nakayashiki H, Nguyen Q B. RNA interference: roles in fungal biology. Curr Opin Microbiol, 2008, 11 (6): 494~502 ［6］ Kim D, Rossi J. RNAi mechanisms and applications. Biotechniques, 2008, 44 (5): 613~616 ［7］ Mine A, Okuno T. Viruses and RNA silencing. Uirusu, 2008, 58 (1): 61~68 ［8］ Dykxhoorn D M, Chowdhury D, Lieberman J. RNA interference and cancer: endogenous pathways and therapeutic approaches. Adv Ex PMed Biol, 2008, 615: 299~329 ［9］ Bernards R, Brummelkam PT R, Beijersbergen R L. shRNA libraries and their use in cancer genetics. Nat Methods, 2006, 3 (9): 701~706 ［10］Lu PY, Woodle M C. Delivering small interfering RNA for novel therapeutics. Methods Mol Biol, 2008, 437: 93~107 ［11］Stogios PJ, Chen L, Privé G G. Crystal structure of the BTB domain from the LRF/ZBTB7 transcriptional regulator. Protein Sci, 2007, 16 (2): 336~342 ［12］Agrawal A, Yang J, Murphy R F, et al. Regulation of the p14ARFMdm2p53 pathway: an overview in breast cancer. Ex PMol Pathol, 2006, 81 (2): 115~122 ［13］Jeon B N, Yoo J Y, Choi W I, et al. Protooncogene FBI1 (Pokemon/ZBTB7A) represses transcription of the tumor suppressor Rb gene via binding competition with Sp1 and recruitment of corepressors. J Biol Chem, 2008, 283 (48): 33199~33210 ［14］Brummelkam PT R, Bernards R, Agami R. A system for stable expression of short interfering RNAs in mammalian cells. Science, 2002, 296 (5567): 550~553 ［15］ Sui G, Soohoo C, Affarel B, et al. A DNA vectorbased RNAi technology to suppress gene expression in mammalian cells. Proc Natl Acad Sci, 2002, 99 (8): 5515~5520 ［16］ Rossi J J. Expression strategies for short hairpin RNA interference triggers. Hum Gene Ther, 2008, 19 (4): 313~317 ［17］Kim D R, Pushparaj PN, Aarthi J J, et al. siRNA, miRNA, and shRNA: in vivo applictions. J Dent Res, 2008, 87 (11): 992~1003 |
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