微小RNA-122在肝癌中的表达与功能研究
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摘要
本文研究了微小RNA-122(miR-122)在原发性肝癌中的表达及其临床意义,并初步探讨了miR-122的靶基因和相应的功能。课题分为两个部分。第一部分:实时荧光定量RT-PCR方法的建立及miR-122与肝癌的关系研究;第二部分miR-122靶分子的确定及功能研究。
首先建立一个特异性强、灵敏度高、重复性好、定量准确、速度快、全封闭的实时定量荧光RT-PCR方法,研究了miR-122在肝癌组织中的表达情况,对肝癌组织和癌旁组织miR-122的表达进行检测。发现与癌旁组织相比,癌肿组织的miR-122表达是下降的。miR-122表达的改变与肿瘤的分化程度有关,也与肿瘤的纤维化程度有关,提示可能作为新的诊断和预后指标。
经生物信息分析,层粘蛋白Y 1亚单位可能是miR-122的靶基因。有两个潜在的结合位点,结合可能性大,自由能低。构建miR-122表达载体和合成miR-122的反义序列(AMO-122),与分离的肝星状细胞作用,从功能增强和抑制两个方面研究其功能。半定量Western Blot分析,miR-122的表达与LAMC1呈负相关,能够通过正、负两个方面调控靶基因LAMC1的表达。
本课题初步探讨了miR-122的表达及与靶基因LAMC1的相互作用,miRNA对HSC的激活不仅与肝纤维化、肝硬化有关,而且与肿瘤发生有关,本研究只是提出了一个设想,从一个全新的角度认识肝硬化和肝癌,为肝癌的发病机制和治疗方法提供新的思路和实验依据。
The objective of this study is to observe the expression and clinical significance of the human microRNA 122 in liver carcinoma, and investigate its target molecule and function. The subject is divided into two parts.
The first part:
To observe the expression and clinical significance of the human microRNA 122 in liver carcinoma by real time flurescent quantitative RT-PCR. We developed a specific, sensitive, reproducible, rapid, accurate and full-sealing real time flurescent quantitative reverse transcriptase polymerase chain reaction (RT-PCR) method. The expression of has-miR-122 was detected in 70 samples of liver carcinoma tissues and those adjacent ones. Compared to the adjacent tissues, the expression of miR-122 is significantly reduced. The low expression of miR-122 is related the neoplasm differentiation degree and liver fibrosis. So it may be look as a new diagnostic and prognosis index.
The second part:
With Pictar and Targetscan software, laminin y 1 may be a target molecule of miR-122. There are two possible pairing positions located in 2490-2496 and 2331-2337 of LAMC1. We constructed miR-122 expression vector and synthesized anti-miRNA oligonucleotides of miR-122(AMO-122). In isolated HSC, it showed lose-of-function and gain-of-function by semi-quantitative western blot. The expression of miR-122 is negatively associated with LAMC1.
Our results investigated the expression of miR-122 and its relationship of LAMC1. Our study will contribute to better understanding of the mechanism of liver carcinoma and be helpful to development of new approaches for cancer therapy.
首先建立一个特异性强、灵敏度高、重复性好、定量准确、速度快、全封闭的实时定量荧光RT-PCR方法,研究了miR-122在肝癌组织中的表达情况,对肝癌组织和癌旁组织miR-122的表达进行检测。发现与癌旁组织相比,癌肿组织的miR-122表达是下降的。miR-122表达的改变与肿瘤的分化程度有关,也与肿瘤的纤维化程度有关,提示可能作为新的诊断和预后指标。
经生物信息分析,层粘蛋白Y 1亚单位可能是miR-122的靶基因。有两个潜在的结合位点,结合可能性大,自由能低。构建miR-122表达载体和合成miR-122的反义序列(AMO-122),与分离的肝星状细胞作用,从功能增强和抑制两个方面研究其功能。半定量Western Blot分析,miR-122的表达与LAMC1呈负相关,能够通过正、负两个方面调控靶基因LAMC1的表达。
本课题初步探讨了miR-122的表达及与靶基因LAMC1的相互作用,miRNA对HSC的激活不仅与肝纤维化、肝硬化有关,而且与肿瘤发生有关,本研究只是提出了一个设想,从一个全新的角度认识肝硬化和肝癌,为肝癌的发病机制和治疗方法提供新的思路和实验依据。
The objective of this study is to observe the expression and clinical significance of the human microRNA 122 in liver carcinoma, and investigate its target molecule and function. The subject is divided into two parts.
The first part:
To observe the expression and clinical significance of the human microRNA 122 in liver carcinoma by real time flurescent quantitative RT-PCR. We developed a specific, sensitive, reproducible, rapid, accurate and full-sealing real time flurescent quantitative reverse transcriptase polymerase chain reaction (RT-PCR) method. The expression of has-miR-122 was detected in 70 samples of liver carcinoma tissues and those adjacent ones. Compared to the adjacent tissues, the expression of miR-122 is significantly reduced. The low expression of miR-122 is related the neoplasm differentiation degree and liver fibrosis. So it may be look as a new diagnostic and prognosis index.
The second part:
With Pictar and Targetscan software, laminin y 1 may be a target molecule of miR-122. There are two possible pairing positions located in 2490-2496 and 2331-2337 of LAMC1. We constructed miR-122 expression vector and synthesized anti-miRNA oligonucleotides of miR-122(AMO-122). In isolated HSC, it showed lose-of-function and gain-of-function by semi-quantitative western blot. The expression of miR-122 is negatively associated with LAMC1.
Our results investigated the expression of miR-122 and its relationship of LAMC1. Our study will contribute to better understanding of the mechanism of liver carcinoma and be helpful to development of new approaches for cancer therapy.
引文
1 J. Chang, E. Nicolas, D. Marks, et al.miR-122, a mammalian liver-specific microRNA, is processed from hcr mRNA and may downregulate the high affinity cationic amino acid transporter CAT-1 [J] .RNA Biol,2004,1 (2): 106-13
2 C. L. Jopling, M. Yi, A. M. Lancaster, et al.Modulation of hepatitis C virus RNA abundance by a liver-specific MicroRNA[J].Science,2005,309(5740): 1577-81
3 L. Gramantieri, M. Ferracin, F. Fornari, et al.Cyclin G1 is a target of miR-122, a microRNA frequently down-regulated in human hepatocellular carcinoma[J].Cancer Res,2007,67(13):6092-9
4 L. F. Sempere, S. Freemantle, I. Pitha-Rowe, et al.Expression profiling of mammalian microRNAs uncovers a subset of brain-expressed microRNAs with possible roles in murine and human neuronal differentiation[J].Genome Biol,2004,5(3):R13
5 T. Babak, W. Zhang, Q. Morris, et al.Probing microRNAs with microarrays: tissue specificity and functional inference[J].Rna,2004,10(11): 1813-9
6 P. Landgraf, M. Rusu, R. Sheridan, et al.A mammalian microRNA expression atlas based on small RNA library sequencing[J].Cell,2007,129(7):1401-14
7 M. A. Feitelson and J. Lee.Hepatitis B virus integration, fragile sites, and hepatocarcinogenesis[J].Cancer Lett,2007,252(2): 157-170
8 C. L. Jopling, K. L. Norman and P. Sarnow.Positive and negative modulation of viral and cellular mRNAs by liver-specific microRNA miR-122[J].Cold Spring Harb Symp Quant Biol,2006,71(369-76
9 Y. Murakami, T. Yasuda, K. Saigo, et al.Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues[J].Oncogene,2006,25(17):2537-45
10 G. A. Calin, C. Sevignani, C. D. Dumitru, et al.Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers[J].Proc Natl Acad Sci U S A,2004,101(9):2999-3004
11 S. Volinia, G. A. Calin, C. G. Liu, et al.A microRNA expression signature of human solid tumors defines cancer gene targets[J].Proc Natl Acad Sci U S A,2006,103(7):2257-61
12 M. V. Iorio, M. Ferracin, C. G. Liu, et al.MicroRNA gene expression deregulation in human breast cancer[J].Cancer Res,2005,65(16):7065-70
13 J. A. Chan, A. M. Krichevsky and K. S. Kosik.MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells[J].Cancer Res,2005,65(14):6029-33
14 C. M. Shachaf, A. M. Kopelman, C. Arvanitis, et al.MYC inactivation uncovers pluripotent differentiation and tumour dormancy in hepatocellular cancer[J].Nature,2004,431(7012):1112-7
15 L. He, X. He, L. P. Lim, et al.A microRNA component of the p53 tumour suppressor network[J] .Nature,2007,447(7148): 1130-4
16 F. Meng, R. Henson, H. Wehbe-Janek, et al.MicroRNA-21 Regulates Expression of the PTEN Tumor Suppressor Gene in Human Hepatocellular Cancer[J].Gastroenterology,2007,133(2):647-58
17 Y. M. Shah, K. Morimura, Q. Yang, et al.Peroxisome proliferator-activated receptor alpha regulates a microRNA-mediated signaling cascade responsible for hepatocellular proliferation[J].Mol Cell Biol,2007,27(12):4238-47
18K. A. O'Donnell, E. A. Wentzel, K. I. Zeller, et al.c-Myc-regulated microRNAs modulate E2Flexpression[J].Nature,2005,435(7043):839-43
19 H. Kutay, S. Bai, J. Datta, et al.Downregulation of miR-122 in the rodent and human hepatocellular carcinomas[J].J Cell Biochem,2006,99(3):671-8
20 J. Krutzfeldt, N. Rajewsky, R. Braich, et al.Silencing of microRNAs in vivo with 'antagomirs'[J].Nature,2005,438(7068):685-9
21 Calin, GA. et al. (2004) MicroRNA profiling reveals distinct signaturesin B cell chronic lymphocytic leukemia. Proc. Natl. Acad. Sci. U. S. A.101,11755-11760
22 Calin, GA. et al. (2005) A microRNA signature associated with prognosis and progression in chronic lymphocytic leukemia. N. Engl.J. Med. 353,1793-1801
23 Metzler, M. et al. (2004) High expression of precursor microRNA-155/BIC RNA in children with Burkitt lymphoma. Genes Chromosomes Cancer 39,167-169
24 Hammond, S.M. et al. (2001) Argonaute 2, a link between genetic and biochemical analysis of RNAi. Science 293, 1146-1150
25 Khvorova, A. et al. (2003) Functional siRNAs and miRNAs exhibit strand bias. Cell 115,209-216
26 Hwang, H.W. and Mendell, J.T. (2004) MicroRNAs cell proliferation, cell death and tumorigenesis. Br. J. Cancer 94,776-780
27 Bartel, D.P. (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 23,281-297
28 He, L. and Hannon, G.J. (2004) MicroRNAs: small RNAs with a big role in gene regulation. Nat. Rev. Genet. 5,522-531
29 Wu, L. et al. (2006) MicroRNAs direct rapid deadenylation of mRNA. Proc. Natl. Acad. Sci. U. S. A. 103,4034-4039
30 Lee, R.C. and Ambros, V. (2001) An extensive class of small RNAs in Caenorhabditis elegans. Science 294, 862-864
31 Lim, L.P. et al. (2003) Vertebrate microRNA genes. Science 299, 1540
32 Lagos Quintana, M. et al. (2001) Identification of novel genes coding for small expressed RNAs. Science 294, 853-858
33 Griffiths-Jones, S. (2004) The microRNA registry. Nucleic Acids Res. 32, D109-D111
34 Berezikov, E. et al. (2005) Philogenetic shadowing and computational identification of human microRNAs. Cell 120,21-24
35 Rajewsky, N. and Socci, N. (2003) Computational identification of microRNA targets. Dev. Biol. 267, 529-535
36 Michael, M.Z. et al. (2003) Reduced accumulation of specific microRNAs in colorectal neoplasia. Mol. Cancer Res. 1,882-891
37 Takamizawa, J. et al. (2004) Reduced expression of the let-7 microRNAs in human lung cancers in association with shortened postoperative survival. Cancer Res. 64,3753-3756
38 Yanaihara, N. et al. (2006) Unique microRNAmolecular profiles in lung cancer diagnosis and prognosis. Cancer Cell 9,189-198
39 Iorio, M.V. et al. (2005) MicroRNA gene expression deregulation in human breast cancer. Cancer Res. 65, 7065-7070
40 Ciafre, S.A. et al. (2005) Extensive modulation of a set of microRNAs in primary glioblastoma. Biochem. Biophys. Res. Commun. 334,1351-1358
41 Volinia, S. et al. (2005) A microRNA expression signature of human solid tumors defines cancer gene targets. Proc. Natl. Acad. Sci. U. S. A. 103,2257-2261
42 He, H. et al. (2005) The role of microRNA genes in papillary thyroid carcinoma. Proc. Natl. Acad. Sci. U.S.A. 102, 19075-19080
43 Felli, N. et al. (2005) MicroRNAs 221 and 222 inhibit normal erithropoiesis and erythroleukemic cell growth via kit receptor down-modulation. Proc. Natl. Acad. Sci. U. S. A. 102, 18081-18086
44 Murakami, Y. et al. (2005) Comprehensive analysis of microRNA expression patterns in hepatocelullar carcinoma and non-tumorous tissues. Oncogene 2005,1-9
1 J.Krutzfeldt,N.Rajewsky,R.Braich,et al.Silencing of microRNAs in vivo with 'antagomirs'[J].Nature,2005,438(7068):685-9
2 R.J.Perera and A.Ray.MicroRNAs in the search for understanding human diseases[J].BioDrugs,2007,21(2):97-104
3 J.Chang,E.Nicolas,D.Marks,et al.miR-122,a mammalian liver-specific microRNA,is processed from hcr mRNA and may downregulate the high affinity cationic amino acid transporter CAT-1[J].RNA Bioi,2004,1(2):106-13
4 C.L.Jopling,K.L.Norman and P.Samow.Positive and negative modulation of viral and cellular mRNAs by liver-specific microRNA miR-122[J].Cold Spring Harb Symp Quant Biol,2006,71(369-76
5 C.L.Jopling,M.Yi,A.M.Lancaster,et al.Modulation of hepatitis C virus RNA abundance by a liver-specific MicroRNA[J].Science,2005,309(5740):1577-81
6 Y.M.Shah,K.Morimura,Q.Yang,et al.Peroxisome proliferator-activated receptor alpha regulates a microRNA-mediated signaling cascade responsible for hepatocellular proliferation[J].Mol Cell Biol,2007,27(12):4238-47
7 Torimura T,Ueno T,Kin M,et al.Coordinated expression of integrin alpha6betal and laminin in hepatocellular carcinoma.Hum Pathol.1997 Oct;28(10):1131-8.
8 张锦生.肝星状细胞激活的内在机制[J].世界华人消化杂志,2005,13(7):831.
9 Everett L.GaIli A.Crabb D.The role of hepatic peroxisome Proliferators activated receptors (PPARs)in health and disease[J].Liver,2000,20:191.
10 L.Gramantieri,M.Ferracin,F.Fornari,et al.Cyclin G1 is a target of miR-122a,a microRNA frequently down-regulated in human hepatocellular carcinoma[J].Cancer Res,2007,67(13):6092-9
1 G.A.Calin and C.M.Croce.MicroRNA-cancer connection:the beginning of a new tale[J].Cancer Res,2006,66(15):7390-4
2 B.Zhang,X.Pan,G.P.Cobb,et al.microRNAs as oncogenes and tumor suppressors[J].Dev Biol,2007,302(1):1-12
3 G.Randall,M.Panis,J.D.Cooper,et al.Cellular cofactors affecting hepatitis C virus infection and replication[J].Proc Natl Acad Sci U S A,2007,104(31):12884-9
4 D.P.Bartel and C.Z.Chen.Micromanagers of gene expression:the potentially widespread influence of metazoan microRNAs[J].Nat Rev Genet,2004,5(5):396-400
5 L.F.Sempere,S.Freemantle,I.Pitha-Rowe,et al.Expression profiling of mammalian microRNAs uncovers a subset of brain-expressed microRNAs with possible roles in murine and human neuronal differentiation[J].Genome Biol,2004,5(3):R13
6 T.Babak,W.Zhang,Q.Morris,et al.Probing microRNAs with microarrays:tissue specificity and functional inference[J].Rna,2004,10(11):1813-9
7 P.Landgraf,M.Rusu,R.Sheridan,et al.A mammalian microRNA expression atlas based on small RNA library sequencing[J].Cell,2007,129(7):1401-14
8 J.Chang,E.Nicolas,D.Marks,et al.miR-122,a mammalian liver-specific microRNA,is processed from her mRNA and may downregulate the high affinity cationic amino acid transporter CAT-1[J].RNA Biol,2004,1(2):106-13
9 M.A.Feitelson and J.Lee.Hepatitis B virus integration,fragile sites,and hepatocarcinogenesis[J].Cancer Lett,2007,252(2):157-170
10 C.L.Jopling,K.L.Norman and P.Sarnow.Positive and negative modulation of viral and cellular mRNAs by liver-specific microRNA miR-122[J].Cold Spring Harb Symp Quant Biol,2006,71(369-76
11 C.L.Jopling,M.Yi,A.M.Lancaster,et al.Modulation of hepatitis C virus RNA abundance by a liver-specific MicroRNA[J].Science,2005,309(5740):1577-81
12 Y.Murakami,T.Yasuda,K.Saigo,et al.Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues[J].Oncogene,2006,25(17):2537-45
13 G.A.Calin,C.Sevignani,C.D.Dumitru,et al.Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers[J].Proc Natl Acad Sci U S A,2004,101(9):2999-3004
14 S. Volinia, G. A. Calin, C. G. Liu, et al. A microRNA expression signature of human solid tumors defines cancer gene targets[J].Proc Natl Acad Sci U S A,2006,103(7):2257-61
15 M. V. Iorio, M. Ferracin, C. G. Liu, et al. MicroRNA gene expression deregulation in human breast cancer[J]. Cancer Res,2005,65(16):7065-70
16 J. A. Chan, A. M. Krichevsky and K. S. Kosik. MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells[J].Cancer Res,2005,65(14):6029-33
17 C.M. Shachaf, A. M. Kopelman, C. Arvanitis, et al. MYC inactivation uncovers pluripotent differentiation and tumour dormancy in hepatocellular cancer[J].Nature,2004,431(7012):1112-7
18 L. He, X. He, L. P. Lim, et al. A microRNA component of the p53 tumour suppressor network[J].Nature,2007,447(7148):1130-4
19 F. Meng, R. Henson, H. Wehbe-Janek, et al. MicroRNA-21 Regulates Expression of the PTEN Tumor Suppressor Gene in Human Hepatocellular Cancer[J].Gastroenterology,2007,133(2):647-58
20 Y. M. Shah, K. Morimura, Q. Yang, et al. Peroxisome proliferator-activated receptor alpha regulates a microRNA-mediated signaling cascade responsible for hepatocellular proliferation[J].Mol Cell Biol,2007,27(12):4238-47
21 K. A. O'Donnell, E. A. Wentzel, K. I. Zeller, et al. c-Myc-regulated microRNAs modulate E2F1 expression[J].Nature,2005,435(7043):839-43
22 L. Gramantieri, M. Ferracin, F. Fornari, et al. Cyclin G1 is a target of miR-122a, a microRNA frequently down-regulated in human hepatocellular carcinoma[J].Cancer Res,2007,67(13):6092-9
23 H. Kutay, S. Bai, J. Datta, et al. Downregulation of miR-122 in the rodent and human hepatocellular carcinomas[J].J Cell Biochem,2006,99(3):671-8
24 J. Krutzfeldt, N. Rajewsky, R. Braich, et al. Silencing of microRNAs in vivo with 'antagomirs'[J].Nature,2005,438(7068):685-9
2 C. L. Jopling, M. Yi, A. M. Lancaster, et al.Modulation of hepatitis C virus RNA abundance by a liver-specific MicroRNA[J].Science,2005,309(5740): 1577-81
3 L. Gramantieri, M. Ferracin, F. Fornari, et al.Cyclin G1 is a target of miR-122, a microRNA frequently down-regulated in human hepatocellular carcinoma[J].Cancer Res,2007,67(13):6092-9
4 L. F. Sempere, S. Freemantle, I. Pitha-Rowe, et al.Expression profiling of mammalian microRNAs uncovers a subset of brain-expressed microRNAs with possible roles in murine and human neuronal differentiation[J].Genome Biol,2004,5(3):R13
5 T. Babak, W. Zhang, Q. Morris, et al.Probing microRNAs with microarrays: tissue specificity and functional inference[J].Rna,2004,10(11): 1813-9
6 P. Landgraf, M. Rusu, R. Sheridan, et al.A mammalian microRNA expression atlas based on small RNA library sequencing[J].Cell,2007,129(7):1401-14
7 M. A. Feitelson and J. Lee.Hepatitis B virus integration, fragile sites, and hepatocarcinogenesis[J].Cancer Lett,2007,252(2): 157-170
8 C. L. Jopling, K. L. Norman and P. Sarnow.Positive and negative modulation of viral and cellular mRNAs by liver-specific microRNA miR-122[J].Cold Spring Harb Symp Quant Biol,2006,71(369-76
9 Y. Murakami, T. Yasuda, K. Saigo, et al.Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues[J].Oncogene,2006,25(17):2537-45
10 G. A. Calin, C. Sevignani, C. D. Dumitru, et al.Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers[J].Proc Natl Acad Sci U S A,2004,101(9):2999-3004
11 S. Volinia, G. A. Calin, C. G. Liu, et al.A microRNA expression signature of human solid tumors defines cancer gene targets[J].Proc Natl Acad Sci U S A,2006,103(7):2257-61
12 M. V. Iorio, M. Ferracin, C. G. Liu, et al.MicroRNA gene expression deregulation in human breast cancer[J].Cancer Res,2005,65(16):7065-70
13 J. A. Chan, A. M. Krichevsky and K. S. Kosik.MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells[J].Cancer Res,2005,65(14):6029-33
14 C. M. Shachaf, A. M. Kopelman, C. Arvanitis, et al.MYC inactivation uncovers pluripotent differentiation and tumour dormancy in hepatocellular cancer[J].Nature,2004,431(7012):1112-7
15 L. He, X. He, L. P. Lim, et al.A microRNA component of the p53 tumour suppressor network[J] .Nature,2007,447(7148): 1130-4
16 F. Meng, R. Henson, H. Wehbe-Janek, et al.MicroRNA-21 Regulates Expression of the PTEN Tumor Suppressor Gene in Human Hepatocellular Cancer[J].Gastroenterology,2007,133(2):647-58
17 Y. M. Shah, K. Morimura, Q. Yang, et al.Peroxisome proliferator-activated receptor alpha regulates a microRNA-mediated signaling cascade responsible for hepatocellular proliferation[J].Mol Cell Biol,2007,27(12):4238-47
18K. A. O'Donnell, E. A. Wentzel, K. I. Zeller, et al.c-Myc-regulated microRNAs modulate E2Flexpression[J].Nature,2005,435(7043):839-43
19 H. Kutay, S. Bai, J. Datta, et al.Downregulation of miR-122 in the rodent and human hepatocellular carcinomas[J].J Cell Biochem,2006,99(3):671-8
20 J. Krutzfeldt, N. Rajewsky, R. Braich, et al.Silencing of microRNAs in vivo with 'antagomirs'[J].Nature,2005,438(7068):685-9
21 Calin, GA. et al. (2004) MicroRNA profiling reveals distinct signaturesin B cell chronic lymphocytic leukemia. Proc. Natl. Acad. Sci. U. S. A.101,11755-11760
22 Calin, GA. et al. (2005) A microRNA signature associated with prognosis and progression in chronic lymphocytic leukemia. N. Engl.J. Med. 353,1793-1801
23 Metzler, M. et al. (2004) High expression of precursor microRNA-155/BIC RNA in children with Burkitt lymphoma. Genes Chromosomes Cancer 39,167-169
24 Hammond, S.M. et al. (2001) Argonaute 2, a link between genetic and biochemical analysis of RNAi. Science 293, 1146-1150
25 Khvorova, A. et al. (2003) Functional siRNAs and miRNAs exhibit strand bias. Cell 115,209-216
26 Hwang, H.W. and Mendell, J.T. (2004) MicroRNAs cell proliferation, cell death and tumorigenesis. Br. J. Cancer 94,776-780
27 Bartel, D.P. (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 23,281-297
28 He, L. and Hannon, G.J. (2004) MicroRNAs: small RNAs with a big role in gene regulation. Nat. Rev. Genet. 5,522-531
29 Wu, L. et al. (2006) MicroRNAs direct rapid deadenylation of mRNA. Proc. Natl. Acad. Sci. U. S. A. 103,4034-4039
30 Lee, R.C. and Ambros, V. (2001) An extensive class of small RNAs in Caenorhabditis elegans. Science 294, 862-864
31 Lim, L.P. et al. (2003) Vertebrate microRNA genes. Science 299, 1540
32 Lagos Quintana, M. et al. (2001) Identification of novel genes coding for small expressed RNAs. Science 294, 853-858
33 Griffiths-Jones, S. (2004) The microRNA registry. Nucleic Acids Res. 32, D109-D111
34 Berezikov, E. et al. (2005) Philogenetic shadowing and computational identification of human microRNAs. Cell 120,21-24
35 Rajewsky, N. and Socci, N. (2003) Computational identification of microRNA targets. Dev. Biol. 267, 529-535
36 Michael, M.Z. et al. (2003) Reduced accumulation of specific microRNAs in colorectal neoplasia. Mol. Cancer Res. 1,882-891
37 Takamizawa, J. et al. (2004) Reduced expression of the let-7 microRNAs in human lung cancers in association with shortened postoperative survival. Cancer Res. 64,3753-3756
38 Yanaihara, N. et al. (2006) Unique microRNAmolecular profiles in lung cancer diagnosis and prognosis. Cancer Cell 9,189-198
39 Iorio, M.V. et al. (2005) MicroRNA gene expression deregulation in human breast cancer. Cancer Res. 65, 7065-7070
40 Ciafre, S.A. et al. (2005) Extensive modulation of a set of microRNAs in primary glioblastoma. Biochem. Biophys. Res. Commun. 334,1351-1358
41 Volinia, S. et al. (2005) A microRNA expression signature of human solid tumors defines cancer gene targets. Proc. Natl. Acad. Sci. U. S. A. 103,2257-2261
42 He, H. et al. (2005) The role of microRNA genes in papillary thyroid carcinoma. Proc. Natl. Acad. Sci. U.S.A. 102, 19075-19080
43 Felli, N. et al. (2005) MicroRNAs 221 and 222 inhibit normal erithropoiesis and erythroleukemic cell growth via kit receptor down-modulation. Proc. Natl. Acad. Sci. U. S. A. 102, 18081-18086
44 Murakami, Y. et al. (2005) Comprehensive analysis of microRNA expression patterns in hepatocelullar carcinoma and non-tumorous tissues. Oncogene 2005,1-9
1 J.Krutzfeldt,N.Rajewsky,R.Braich,et al.Silencing of microRNAs in vivo with 'antagomirs'[J].Nature,2005,438(7068):685-9
2 R.J.Perera and A.Ray.MicroRNAs in the search for understanding human diseases[J].BioDrugs,2007,21(2):97-104
3 J.Chang,E.Nicolas,D.Marks,et al.miR-122,a mammalian liver-specific microRNA,is processed from hcr mRNA and may downregulate the high affinity cationic amino acid transporter CAT-1[J].RNA Bioi,2004,1(2):106-13
4 C.L.Jopling,K.L.Norman and P.Samow.Positive and negative modulation of viral and cellular mRNAs by liver-specific microRNA miR-122[J].Cold Spring Harb Symp Quant Biol,2006,71(369-76
5 C.L.Jopling,M.Yi,A.M.Lancaster,et al.Modulation of hepatitis C virus RNA abundance by a liver-specific MicroRNA[J].Science,2005,309(5740):1577-81
6 Y.M.Shah,K.Morimura,Q.Yang,et al.Peroxisome proliferator-activated receptor alpha regulates a microRNA-mediated signaling cascade responsible for hepatocellular proliferation[J].Mol Cell Biol,2007,27(12):4238-47
7 Torimura T,Ueno T,Kin M,et al.Coordinated expression of integrin alpha6betal and laminin in hepatocellular carcinoma.Hum Pathol.1997 Oct;28(10):1131-8.
8 张锦生.肝星状细胞激活的内在机制[J].世界华人消化杂志,2005,13(7):831.
9 Everett L.GaIli A.Crabb D.The role of hepatic peroxisome Proliferators activated receptors (PPARs)in health and disease[J].Liver,2000,20:191.
10 L.Gramantieri,M.Ferracin,F.Fornari,et al.Cyclin G1 is a target of miR-122a,a microRNA frequently down-regulated in human hepatocellular carcinoma[J].Cancer Res,2007,67(13):6092-9
1 G.A.Calin and C.M.Croce.MicroRNA-cancer connection:the beginning of a new tale[J].Cancer Res,2006,66(15):7390-4
2 B.Zhang,X.Pan,G.P.Cobb,et al.microRNAs as oncogenes and tumor suppressors[J].Dev Biol,2007,302(1):1-12
3 G.Randall,M.Panis,J.D.Cooper,et al.Cellular cofactors affecting hepatitis C virus infection and replication[J].Proc Natl Acad Sci U S A,2007,104(31):12884-9
4 D.P.Bartel and C.Z.Chen.Micromanagers of gene expression:the potentially widespread influence of metazoan microRNAs[J].Nat Rev Genet,2004,5(5):396-400
5 L.F.Sempere,S.Freemantle,I.Pitha-Rowe,et al.Expression profiling of mammalian microRNAs uncovers a subset of brain-expressed microRNAs with possible roles in murine and human neuronal differentiation[J].Genome Biol,2004,5(3):R13
6 T.Babak,W.Zhang,Q.Morris,et al.Probing microRNAs with microarrays:tissue specificity and functional inference[J].Rna,2004,10(11):1813-9
7 P.Landgraf,M.Rusu,R.Sheridan,et al.A mammalian microRNA expression atlas based on small RNA library sequencing[J].Cell,2007,129(7):1401-14
8 J.Chang,E.Nicolas,D.Marks,et al.miR-122,a mammalian liver-specific microRNA,is processed from her mRNA and may downregulate the high affinity cationic amino acid transporter CAT-1[J].RNA Biol,2004,1(2):106-13
9 M.A.Feitelson and J.Lee.Hepatitis B virus integration,fragile sites,and hepatocarcinogenesis[J].Cancer Lett,2007,252(2):157-170
10 C.L.Jopling,K.L.Norman and P.Sarnow.Positive and negative modulation of viral and cellular mRNAs by liver-specific microRNA miR-122[J].Cold Spring Harb Symp Quant Biol,2006,71(369-76
11 C.L.Jopling,M.Yi,A.M.Lancaster,et al.Modulation of hepatitis C virus RNA abundance by a liver-specific MicroRNA[J].Science,2005,309(5740):1577-81
12 Y.Murakami,T.Yasuda,K.Saigo,et al.Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues[J].Oncogene,2006,25(17):2537-45
13 G.A.Calin,C.Sevignani,C.D.Dumitru,et al.Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers[J].Proc Natl Acad Sci U S A,2004,101(9):2999-3004
14 S. Volinia, G. A. Calin, C. G. Liu, et al. A microRNA expression signature of human solid tumors defines cancer gene targets[J].Proc Natl Acad Sci U S A,2006,103(7):2257-61
15 M. V. Iorio, M. Ferracin, C. G. Liu, et al. MicroRNA gene expression deregulation in human breast cancer[J]. Cancer Res,2005,65(16):7065-70
16 J. A. Chan, A. M. Krichevsky and K. S. Kosik. MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells[J].Cancer Res,2005,65(14):6029-33
17 C.M. Shachaf, A. M. Kopelman, C. Arvanitis, et al. MYC inactivation uncovers pluripotent differentiation and tumour dormancy in hepatocellular cancer[J].Nature,2004,431(7012):1112-7
18 L. He, X. He, L. P. Lim, et al. A microRNA component of the p53 tumour suppressor network[J].Nature,2007,447(7148):1130-4
19 F. Meng, R. Henson, H. Wehbe-Janek, et al. MicroRNA-21 Regulates Expression of the PTEN Tumor Suppressor Gene in Human Hepatocellular Cancer[J].Gastroenterology,2007,133(2):647-58
20 Y. M. Shah, K. Morimura, Q. Yang, et al. Peroxisome proliferator-activated receptor alpha regulates a microRNA-mediated signaling cascade responsible for hepatocellular proliferation[J].Mol Cell Biol,2007,27(12):4238-47
21 K. A. O'Donnell, E. A. Wentzel, K. I. Zeller, et al. c-Myc-regulated microRNAs modulate E2F1 expression[J].Nature,2005,435(7043):839-43
22 L. Gramantieri, M. Ferracin, F. Fornari, et al. Cyclin G1 is a target of miR-122a, a microRNA frequently down-regulated in human hepatocellular carcinoma[J].Cancer Res,2007,67(13):6092-9
23 H. Kutay, S. Bai, J. Datta, et al. Downregulation of miR-122 in the rodent and human hepatocellular carcinomas[J].J Cell Biochem,2006,99(3):671-8
24 J. Krutzfeldt, N. Rajewsky, R. Braich, et al. Silencing of microRNAs in vivo with 'antagomirs'[J].Nature,2005,438(7068):685-9
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