靶向MMP-9的脱氧核酶对人肺腺癌A549细胞粘附与迁移作用的影响
|
摘要
第一部分靶向MMP-9的脱氧核酶抑制人肺腺癌A549细胞中MMP-9蛋白表达
目的
探讨靶向MMP-9的脱氧核酶(DNAzyme,Deoxyri bozyme)对人肺腺癌A549细胞中MMP-9蛋白表达的影响。确定靶向MMP-9的脱氧核酶对MMP-9的mRNA切割的有效性。
方法
设计靶向MMP-9的脱氧核酶,应用oligofectamine将脱氧核酶转染到A549细胞中。实验分三组:空白组为A549培养细胞,对照组为将control oligo转染A549细胞,实验组为靶向MMP-9的脱氧核酶转染A549细胞。靶向MMP-9的脱氧核酶与对照control oligo终浓度均为2 uM。采用Western Blot方法检测细胞中MMP-9蛋白表达。
结果
在靶向MMP-9的脱氧核酶干预1d后人肺腺癌A549细胞中MMP-9蛋白表达较对照组与空白组显著减低(p＜0.01),而空白组与对照组细胞中蛋白条带灰度值无明显差异(p＞0.05)。
结论
靶向MMP-9的脱氧核酶在肺腺癌A549细胞中能够有效地抑制MMP-9蛋白的高表达。
第二部分靶向MMP-9的脱氧核酶对人肺腺癌A549细胞粘附与迁移作用的影响
目的
在证实靶向MMP-9的脱氧核酶能够有效抑制A549细胞中MMP-9蛋白表达的基础上,我们进一步研究靶向MMP-9的脱氧核酶对人肺腺癌A549细胞粘附与迁移作用的影响。
方法
应用oligofectamine将脱氧核酶转染到A549细胞中。利用细胞粘附与迁移试验观察脱氧核酶干预后细胞粘附与迁移能力的变化。
结果
经靶向MMP-9的脱氧核酶干预后,2h和4h细胞粘附率,迁移速度均显著降低(p＜0.01),而对照组与空白组之间无差异(P＞0.05)。
结论
靶向MMP-9的脱氧核酶在肺腺癌A549细胞中能够有效阻止细胞的粘附与迁移。
Objective
To investigate the effect of targeting MMP-9 deoxyribozyme on expression of MMP-9 in human lung adenocarcinoma cell line A549, then determine the effectiveness of targeting MMP-9 deoxyribozyme on cleavage of mRNA in MMP-9.
Methods
Designed the targeting MMP-9 deoxyribozyme, transfected deoxyribozyme into human lung adenocarcinoma cell line A549, applied oligofectamine method. Experiment were divided into 3 groups: empty group: The human lung adenocarcinoma cell line A549. Control group: Transfected control oligo into human lung adenocarcinoma cell line A549. Experimental group: Tansfected targeting MMP-9 deoxyribozyme into human lung adenocarcinoma cell line A549. The concentrations of targeting MMP-9 deoxyribozyme and control oligo are 2 umol/L. Using western blot method to detect the expression of MMP-9 in cell.
Results
After targeting MMP-9 deoxyribozyme intervention one day, the expression of MMP-9 in the cells compared with the control group was significantly lower (p<0.01). There was no difference between the empty group and the control group in gradation of protein strap (p>0.05).
Conclusions
Targeting MMP-9 deoxyribozyme effectively inhibited the expression of MMP-9 in human lung adenocarcinoma cell line A549.
Objective
To investigate the effect of targeting MMP-9 deoxyribozyme on cell adhesion and migration in human lung adenocarcinoma cancer cell line A549.
Methods
Designed the targeting MMP-9 deoxyribozyme, transfected deoxyribozyme into human lung adenocarcinoma cell line A549, applied oligofectamine method. To observa the cell adhesion and migration after the intervention of deoxyribozyme.
Results
After targeting MMP-9 deoxyribozyme interventioned, the rate of cell adhesion and migration were significantly decreased (p<0.01). There was no difference between the empty group and the control group (p<0.05).
Conclusions
Targeting MMP-9 deoxyribozyme effectively prevent cell adhesion and migration.
目的
探讨靶向MMP-9的脱氧核酶(DNAzyme,Deoxyri bozyme)对人肺腺癌A549细胞中MMP-9蛋白表达的影响。确定靶向MMP-9的脱氧核酶对MMP-9的mRNA切割的有效性。
方法
设计靶向MMP-9的脱氧核酶,应用oligofectamine将脱氧核酶转染到A549细胞中。实验分三组:空白组为A549培养细胞,对照组为将control oligo转染A549细胞,实验组为靶向MMP-9的脱氧核酶转染A549细胞。靶向MMP-9的脱氧核酶与对照control oligo终浓度均为2 uM。采用Western Blot方法检测细胞中MMP-9蛋白表达。
结果
在靶向MMP-9的脱氧核酶干预1d后人肺腺癌A549细胞中MMP-9蛋白表达较对照组与空白组显著减低(p＜0.01),而空白组与对照组细胞中蛋白条带灰度值无明显差异(p＞0.05)。
结论
靶向MMP-9的脱氧核酶在肺腺癌A549细胞中能够有效地抑制MMP-9蛋白的高表达。
第二部分靶向MMP-9的脱氧核酶对人肺腺癌A549细胞粘附与迁移作用的影响
目的
在证实靶向MMP-9的脱氧核酶能够有效抑制A549细胞中MMP-9蛋白表达的基础上,我们进一步研究靶向MMP-9的脱氧核酶对人肺腺癌A549细胞粘附与迁移作用的影响。
方法
应用oligofectamine将脱氧核酶转染到A549细胞中。利用细胞粘附与迁移试验观察脱氧核酶干预后细胞粘附与迁移能力的变化。
结果
经靶向MMP-9的脱氧核酶干预后,2h和4h细胞粘附率,迁移速度均显著降低(p＜0.01),而对照组与空白组之间无差异(P＞0.05)。
结论
靶向MMP-9的脱氧核酶在肺腺癌A549细胞中能够有效阻止细胞的粘附与迁移。
Objective
To investigate the effect of targeting MMP-9 deoxyribozyme on expression of MMP-9 in human lung adenocarcinoma cell line A549, then determine the effectiveness of targeting MMP-9 deoxyribozyme on cleavage of mRNA in MMP-9.
Methods
Designed the targeting MMP-9 deoxyribozyme, transfected deoxyribozyme into human lung adenocarcinoma cell line A549, applied oligofectamine method. Experiment were divided into 3 groups: empty group: The human lung adenocarcinoma cell line A549. Control group: Transfected control oligo into human lung adenocarcinoma cell line A549. Experimental group: Tansfected targeting MMP-9 deoxyribozyme into human lung adenocarcinoma cell line A549. The concentrations of targeting MMP-9 deoxyribozyme and control oligo are 2 umol/L. Using western blot method to detect the expression of MMP-9 in cell.
Results
After targeting MMP-9 deoxyribozyme intervention one day, the expression of MMP-9 in the cells compared with the control group was significantly lower (p<0.01). There was no difference between the empty group and the control group in gradation of protein strap (p>0.05).
Conclusions
Targeting MMP-9 deoxyribozyme effectively inhibited the expression of MMP-9 in human lung adenocarcinoma cell line A549.
Objective
To investigate the effect of targeting MMP-9 deoxyribozyme on cell adhesion and migration in human lung adenocarcinoma cancer cell line A549.
Methods
Designed the targeting MMP-9 deoxyribozyme, transfected deoxyribozyme into human lung adenocarcinoma cell line A549, applied oligofectamine method. To observa the cell adhesion and migration after the intervention of deoxyribozyme.
Results
After targeting MMP-9 deoxyribozyme interventioned, the rate of cell adhesion and migration were significantly decreased (p<0.01). There was no difference between the empty group and the control group (p<0.05).
Conclusions
Targeting MMP-9 deoxyribozyme effectively prevent cell adhesion and migration.
引文
[1]Kawai H,Kiura K,Tabata M,et al.Characterization of non-small-cell lung Cancer cell lines established before and after Chemotherapy.Lung Cancer,2002,35(3):305-314.
[2]Fong km,kida Y,Zimmerran PV,et al.TIMP1 and adverse Prog-nosis innon-samll-cell lung Cancar.Clin Cancer Res,1996,2(8):1369-1372
[3]Karameris A,Panagou P,T silais T,et al.Association of expression of finefall-oproteinases and their inhibitors with the metastatic potential of Squamous-cell lung Carcinomas A molecular and immunohis to chemical Study.Am J Respir Crit Care Med,1997,156(6):1903-1936.
[4]Deryugina El,Quigley JP.Matrix metalloproteinases and tumor metastasis[J].Cancer Metastasis Rev.2006,25(1):9-34.
[5]Jukka W.,Veli-Matti K.Regulation of matrix metallpproteinase expression in tumor invasion.J FASEB 1999,13:781-792.
[6]Bruce P.H,Edward j.L,Hiroshi sato,et al.transcriptional activation of the matrix metalloproteinase-9 gene in an H-ras and v-myc transformed rat embryo cell line.Oncogene,1997,14:1995-1998.
[7]R.Gum,H Wang,E Lengyel,et al.Regulation of 92 kDa type IV collagenase expression by the jun aminoterminal kinase and the extracellular signal-regulated kinase dependent signaling cascades.Oncogene,1997,14:1481-1493.
[8]Hajime T,Tomokazu Y,William E.M,et al.Matrix Metalloproteinase 9expression Islnduced by epstein-Barr Virus Latent Membrane 1 C-Terminal Activation Regions land 2[J].Journal of Virology,1999,73(7):5548-5555.
[9]Toshiyuki H.,Tomokazu Y.,Tzung-Shiahn S.et al.Association of latent membrane protein 1 and matrix metalloproteinase 9 with metastasis in nasopharyngeal carcinoma.Cancer,2000,89(4):715-723.
[10]Tomokazu Y,Hiroshi Sato,Mitsuru E et al.The expression of matrix metalloproteinase 9 is enhanced by Epstein-Barr virus latent membrane protein 1.Proc.Natl.Acad.USA 1998,95:3621-3626.
[11]Madigan MC,Kingsley EA,Cozzi PJ,et al.The role of extracellular matrix metalloproteinase inducer protein in prostate cancer progression[J].Cancer Immunol Immunother.2008,Feb 14[Epub ahead of print]
[12]Rollin J,Regina S,Vourch P,et al.Influence of MMP-2 and MMP-9promoter polymorphisms on gene expression and clinical outcome of non-small cell lung cancer.Lung cancer,2007 May;56(2):273-80.
[13]Ko HM,Kang JH,Jung B,et al.Critical role for matrix metalloproteinase-9 in platelet-activating factor-induced experimental tumor metastasis.Int J Cancer,2007 Mar 15;126(6):1277-83.
[14]Taras D,Blanc JF,Rullier A,et al.Pravastatin reduces lung metastasis of rat hepatocellular carcinoma via a coordinated decrease of MMP expression and activity.J Hepatol,2007 Jan;46(1):69-76.
[15]谢建武,方伟岗,李红梅等,金属蛋白酶MMP-9反义基因对高转移性肺巨细胞癌系的转染效应.中华病理学杂志,1999,28(4):210-211.
[16]牛桂英,方伟岗,郑杰等,TIMP-3基因转染抑制人肺癌细胞系浸润与转移的研究.中华病理学杂志,1998,27(6):421-424.
[17]Trulzsch B,Wood M.Applications of nucleic acid technology in the CNS.J Neurochem,2004;88(2):257-265.
[18]Finkel E.DNA cuts its teeth—as an enzyme.Science,1999;286:2441-2442.
[19]Kurreck J,Bieber B,Jahnel R,et al.Comparative study of DNA enzymes and ribozymes against the same full-length messenger RNA of the vanilloid receptor subtype I.J Biol Chem,2002;277(9):7099-7107.
[20]Breaker RR and Joyce GF.A DNA enzyme that cleaves RNA.Chem.Biol.1994;1(4):223-229.
[21]卢忠心,曹亚.脱氧核酶及其应用进展.国外医学分子生物学分册,2003;25(5):282-5.
[22]Santoro SW,Joyce GF,A general purpose RNA-cleaving DNAenzyme[J].Pro Nat Acad sci USA.1997,94:4262-4266.
[23]Unwalla H,Baner jea AC.Novel Mono-and-di-DNA-enzymes Targeted to Cleave TAT or TAT-REV RNA inhibit HIV-1 Gene Expression[J],Antiviral Res,2001,51(2):127-139.Application[J].Nucleosiddes Nuclweic Acid,2003,22(5):601-604.
[24]Wengle J,Vestev B.LNA and Alpha-L-LNA=TO wards Therapeutic Application[J].Nucleosiddes Nuclweic Acid,2003,22(5):601-604.
[25]Sioud M,Leirdal M,Dedsign of Nucles Resistant Protein Kinas Ca DNA Enzymes with potential therapelltic Application[J],.Mol Biol 2000,296(3):937-947.
[26]Warashine M,Kuwabara T,Nadamatsu Y,et al.Extremelly High and specipic Activity of DNA Enzymes in cells with philadelphiia Chromosome[J].Chem Biol,1999,6(4):237-250.
[27]于乐成,王升启,顾长海等,脱氧核酶再转基因肝癌细胞中对HCV-5非编码区的抑制活性[J],中华传染病杂志,2004,22(2):82-85.
[28]Yen L,Strittmatter SM,Kalb RG.Sequence-specific cleavage of Huntingtin mRNA by catalytic DNA[J].Ann Neurol,1999,46(3):366.
[29]Khachigian LM,Fahmy RG,Zhang G.et al.c-Jun regulates vascular smooth muscle cell growth and neointima formation after arterial injury.Inhibition by a novel DNA enzyme targeting c-Jun[J].J Biol Chem,2002,277(25):22985.
[30]Lowe HC,Chesterman CN,Khachigian LM.Catalytic antisense DNA molecules targeting Egr-1 inhibit neointima formation following permanent ligation of rat common carotid arteries[J].Thromb Haemost,2002,87(1):134.
[31]Dass CR,Choong PF,Khachigian LM.DNAzyme technology and cancer therapy:cleave and let die[J].Mol Cancer Ther.2008,7(2):243-51.
[32]Sasagawa T,Shimakage M,Nakamura M,et al.Epstein-Barr virus(EBV)genes expression in cervical intraepithelial neoplasia and invasive cervical cancer:a comparative study with human papilloma virus(HPV)infection.Hum Pathol.2000;31(3):318-326.
[33]Jen KY,Cheng J,Li J,et al.Mutational events in LMP1 gene of Epstein-Barr virus in salivary gland lymphoepithelial carcinomas.Int J Cancer.2003; 105(5):654-660.
[34]Kuo T,Hsueh C.Lymphoepithelioma-like salivary gland carcinoma in Taiwan:a clinicopathological study of nine cases demonstrating a strong association with Epstein-Barr virus.Histopathology.1997;31(1):75-82.
[35]Morbini P,Riboni R,Tomaselli S,et al.Eber- and LMP-l-expressing pulmonary lymphoepithelioma-like carcinoma in a Caucasian patient.Hum Pathol.2003;34(6):623-5.
[36]Lu ZX,Ma XQ,Yang LF,Wang ZL,DNAzymes targeted to EBV-encoded latent membrane protein-1 induce apoptosis and enhance radiosensitivity in nasopharyngeal carcinoma.Cancer Lett.2008 Mar 17.
[37]Rao JS,Gondi C,Chetty C,et al.Inhibition of invasion,angiogenesis,tumor growth,and metastasis by adenovirus-mediated transfer of antisense uPAR and MMP-9 in non-small cell lung cancer cells[J].Mol Cancer Ther.2005,4(9):1399-408.
[38]Tummalapalli P,Spomar D,Gondi CS,et al.RNAi-mediated abrogation of cathepsin B and MMP-9 gene expression in a malignant meningioma cell line leads to decreased tumor growth,invasion and angiogenesis[J].Int J Oncol.2007,31(5):1039-50.
[39]Tummalapalli P,Spomar D,Gondi CS,et al.RNAi-mediated abrogation of cathepsin B and MMP-9 gene expression in a malignant meningioma cell line leads to decreased tumor growth,invasion and angiogenesis[J].Int J Oncol.2007,31(5):1039-50.
[40]Nakamura T,Kuwai T,Kim JS,et al.Stromal metalloproteinase-9 is essential to angiogenesis and progressive growth of orthotopic human pancreatic cancer in parabiont nude mice[J].Neoplasia.2007,9(11):979-86.
[41]Zheng H,Takahashi H,Murai Y,et al.Expressions of MMP-2,MMP-9 and VEGF are closely linked to growth,invasion,metastasis and angiogenesis of gastric carcinoma[J].Anticancer Res.2006,26(5A):3579-83.
[42]藤博,辛丁,文莲姬等。10-23脱氧核酶靶向抑制喉癌eIF4E基因表达的实 验研究[J].临床耳鼻咽喉头颈外科杂志,2007,21(12):552-4.
[43]Zhang L,Gasper WJ,Stass SA,et al.Angiogenic inhibition mediated by a DNAzyme that taygets vascular endothelial growth factor receptor[J].Cancer Research,2002,62(19):5463-5469.
[44]Santoro SW,Joyce GF.Mechanism and utility of an RNA-cleaving DNA enzyme.Biochemistry.1998 Sep 22;37(38):13330-42.
[45]Cairns M J,Hopkins TM,Witherington C,et al.The influence of arm length asymmetry and base substitution on the activity of the 10-23 DNA enzyme.Antisense Nucleic Acid Drug Dev.2000;10(5):323-32.
[46]Pan WH,Devlin HF,Kelley C,et al.A selection system for identifying accessible sites in target RNAs.RNA.2001;7(4):610-21.
[47]Cairns M J,Hopkins TM,Witherington C,et al.Target site selection for an RNA-cleaving catalytic DNA.Nat Biotechnol.1999;17(5):480-6.
[48]Yen L,Strittmatter SM,Kalb RG.Sequence-specific cleavage of Huntingtin mRNA by catalytic DNA.Ann Neurol.1999;46(3):366-73.
[1] Ray M, Stetier-Stevenson W GGelatinase A activity directly modulatesmelanoma cell adhesion and spreading [J] EMBO J, 1995,14, (4):908-917
[2] Pereda MP,Hopfner U,Pagotto U,et al.Retinoic acid stinulates meningioma cell adhesion to the extracellumatrix and inhibits invasion [J],BrJ Cancer, 1999, 81(3):381-386
[3] Ueda M,Terai Y, Kumagai k, et al.Correlation between thymidine phosphorylase expression and invasion phenotype in cervical carcinoma cell [J], Int J Cancer, 2001, 91(6):778-782
[4] Masahito T, Jianguo G, Huan T, et al.PTEN gene and integrin signaling in cancer[J]. Journal of the Nation cancer Institute,1999,91(2):1820.
[5] Yang Y, Dang D, A takilit A,et al.Specific alpha vintegrin receptors modulate K1735 murine melanoma cell behavior [J],Bio Chem Biophys Res Commun, 2003, 308(4):814-819
[6] Clezardin P. Recent insights ioto the role of inegrins in cancer Metastasis[J]. Mol. Life. Sic, 1998, 54:541
[7] Grant MB, Caballero S,Kornberg L,et al. Focal Adhesion Kinase(FAK) Overexpression Modulates Retinal Angiogenesis [J] .Invest Ophthalmol Vis Sic, 2004,45:4463-4469.
[8] Gou XM, Chen XY, Arrand JR, et al. Effects of Epstein-Barr virus la tent membrane proteinl ( EBV-LMP1 ) on related factors of metastasis of nasopharyngeal carcinoma cell line CNE1 [J]. Ai Zheng, 2003, 22(5):481-485.
[9] Guishui Zhang, Crispin R. Dass, Eric Sumithran, et al. Effect of Deoxyribozymes Targeting c-Jun on Solid Tumor Growth and Angiogenesis in Rodents[J]. Journal of the National Cancer Institute, 2004, 96(9):683-696.
[10] Pasco S, Ramont L, Maquart FX, et al. Control of melanoma progression by various matriknes from basement membrane macromolecules [J]. Crit Rev Oncol Hematol, 2004,49:221-233.
[11] Eibl RH, Benoit M. Molecular resolution of cell adhesionforces[J].IEE Proc Nanobiotechnol.2004 Jun;151(3):128-32.
[12]Tamura M,Gu J,Matsumoto k,et al.Inhibition of cell migration,and focal adhesion by tumor suppressor PTEN[].Science,1998,280:1614.
[13]Pukac L,Huangpu J,Karnovsky MJ.Platelet-drived growth factor-BB,insulin-link growth factor-1,and phorbol ester activate different signaling pathway for stimulation of vascular smooth muscle cell migration[J].Exp.Cell.Res,1998,242:548.
[14]Sheppard D,Characterization of the integrin avβ6 as a fibronectin-binding protein[J].J.Bio.Chem,1992,267:5790.
[15]方新初,苏剑敏,周国非等。整合蛋白a5,β1亚基的过表达对肝癌细胞粘附与迁移的影响[J]。中国癌症杂志,2001,11(1):1-4。
[16]Taras D,Blanc JF,Rullier A,et al.Pravastatin reduces lung metastasis of rat hepatocellular carcinoma via a coordinated decrease of MMP expression and activity.J Hepatol,2007 Jan;46(1):69-76.
[17]Ko HM,Kang JH,Jung B,et al.Critical role for matrix metalloproteinase-9 in platelet-activating factor-induced experimental tumor metastasis.Int J Cancer,2007 Mar 15;126(6):1277-83.
[1]Santoro S W,Joyce GF.A general purpose RNA-cleaving DNA enzyme[J].Proc Natl Acad Sci USA,1997,94(9):4262
[2]Unwalla H,Baner jea AC.Novel Mono-and-di-DNA-enzymes Targeted to Cleave TAT or TAT-REV RNA inhibit HIV-1 Gene Expression[J],Antiviral Res,2001,51(2):127-139.Application[J].Nucleosiddes Nuclweic Acid,2003,22(5):601-604.
[3]Wengle J,Vestev B.LNA and Alpha-L-LNA=TO wards Therapeutic Application [J].Nucleosiddes Nuclweic Acid,2003,22(5):601-604.
[4]Sioud M,Leirdal M,Dedsign of Nucles Resistant Protein Kinas Ca DNA Enzymes with potential therapelltic Application[J],Mol Biol 2000,296(3):937-947.
[5]Warashine M,Kuwabara T,Nadamatsu Y,et al.Extremelly High and specipic Activity of DNA Enzymes in cells with philadelphiia Chromosome[J].Chem Biol,1999,6(4):237-250.
[6]于乐成,王升启,顾长海等,脱氧核酶再转基因肝癌细胞中对HCV-5非编码区的抑制活性[J],中华传染病杂志,2004,22(2):82-85.
[7]Yen L,Strittmatter SM,Kalb RG.Sequence-specific cleavage of Huntingtin mRNA by catalytic DNA[J].Ann Neurol,1999,46(3):366.
[8]Khachigian LM,Fahmy RG,Zhang G.et al.c-Jun regulates vascular smooth muscle cell growth and neointima formation after arterial injury.Inhibition by a novel DNA enzyme targeting c-Jun[J].J Biol Chem,2002,277(25):22985.
[9]Lowe HC,Chesterman CN,Khachigian LM.Catalytic antisense DNA molecules targeting Egr-1 inhibit neointima formation following permanent ligation of rat common carotid arteries[J].Thromb Haemost,2002,87(1):134.
[10]Emilsson GM,Breaker RR.Deoxyribozymes:new activities and new applications[J].Cell Mol Life Sci,2002,59(4):596
[11]Zuker M.Mfold web server for nucleic acid folding and hybridization prediction.Nucleic Acids Res.2003;31(13),3406-15.
[12]Zaborowska Z,Furste JP,Erdmann VA,et al.Sequence requirements in the catalytic core of the "10-23" DNA enzyme.J Biol Chem.2002;277(43):40617-22.
[13]Zaborowska Z,Schubert S,Kurreck J,et al.Deletion analysis in the catalytic region of the 10-23 DNA enzyme.FEBS Lett.2005;579(2):554-8.
[14]Cairns MJ,King A,Sun LQ.Optimisation of the 10-23 DNAzyme-substrate pairing interactions enhanced RNA cleavage activity at purine-cytosion target sites[J].Nucleic Acids Res,2003,31(11):2883-2889.
[15]Cairns MJ,Hopkins TM,Witherington C,et al.Target site selction for an RNA-cleaving catalytic DNA[J].Nat Biotechnol,1999,17(5):480-486
[16]Cairns MJ,Hopkins TM,witherington C,et al,The influence of arm length asymmetry and base substitution on the activity of the 10-23DNA enzyme[J].Antisense Nuleic Acid Drug Dev,2000,10(5):323-332
[17]Yen C,strittmater SM,kalb,RG.sequence-specific cleanage of Huntingon mRNA by Catalytic DNA[J].Ann Neurol,1999,46(3):366-373
[18]Sioud M,leirdal M.Design of nuclease resistant protein kinase k alpha DNA enzyme with potential therapeutic application[J].J Mol Bid,2000,296(3):937-947
[19]Asahine y,lto y,wu CH,et al.DNA ribonucleases that are active against intracellular hepatitis B viral RNA targets[J].Hepatology,1998,28(2):547-554.
[21]Vester B,Lundberg LB,sorensen MD,et al,lmprovd RNA cleavage by LNAzyme derivatives of DNAzymes[J].Biochen soc Trans,2004,32(p+1):37-40.
[22]Kubo T,Takamon K.Conjugate DNAzymes[J].Nucleic Acid Res,2003,3(Suppl):177-178
[23]Zaborowska 2,Furste JP,Erdmann VA.etal.Sequence requirements in the catalytic core of the "10-23"DNA enzyme[J].J Biol chem,2002,277(43):40617-40622
[24]Abdelqang A,wood M,Beeson D.Hairpin DNAzymes:a new tool for efficient cellular gene silencing[J].Gene Med,2007,Aug;9(8):727-738
[25] Jakobsen MR,Haasnoot J, Wenqel J, et al.Eefficient inhibition of HIV-1 expression by LNA modiffied antisense digonucleotides and DNAzymes targeted to functionlly selected binding sites, Retrovirology, 2007, Apr, 26(4):29
[26] Zhang L,Gasper WJ, stass SA, et al .Angiogenic inhibition mediated by a DNAzyme that targets Vascular endothelial growth factor receptor 2[J], Cancer Research, 2002, 62(19):5463-5469.
[27] Liu C, cheng R,Sun LQ, et al.Suppression of platelettype 12-lipoxygenase activity in human erythroleuke miacells by an KNA-cleaving DNAzyme[J], Biochem Biophys Res com, 2001.284(4):1077
[28] Wu Y, Yu L,Mcmahon R,et al.Inhibition of bcr-abl oncogen expression by novel dexyribozymes(DNAzymes)[J].Humei Gene Ther,1999,10(17):2847-2857.
[29] Liang Z, Wei S,Guan J, et al.DNAzyme mediated cleavage of surviving mRNA and inhibition of the growth of PANC-1 cell[J]. J Gastroenterol Hepatol, 2005, 20(10):1595
[2]Fong km,kida Y,Zimmerran PV,et al.TIMP1 and adverse Prog-nosis innon-samll-cell lung Cancar.Clin Cancer Res,1996,2(8):1369-1372
[3]Karameris A,Panagou P,T silais T,et al.Association of expression of finefall-oproteinases and their inhibitors with the metastatic potential of Squamous-cell lung Carcinomas A molecular and immunohis to chemical Study.Am J Respir Crit Care Med,1997,156(6):1903-1936.
[4]Deryugina El,Quigley JP.Matrix metalloproteinases and tumor metastasis[J].Cancer Metastasis Rev.2006,25(1):9-34.
[5]Jukka W.,Veli-Matti K.Regulation of matrix metallpproteinase expression in tumor invasion.J FASEB 1999,13:781-792.
[6]Bruce P.H,Edward j.L,Hiroshi sato,et al.transcriptional activation of the matrix metalloproteinase-9 gene in an H-ras and v-myc transformed rat embryo cell line.Oncogene,1997,14:1995-1998.
[7]R.Gum,H Wang,E Lengyel,et al.Regulation of 92 kDa type IV collagenase expression by the jun aminoterminal kinase and the extracellular signal-regulated kinase dependent signaling cascades.Oncogene,1997,14:1481-1493.
[8]Hajime T,Tomokazu Y,William E.M,et al.Matrix Metalloproteinase 9expression Islnduced by epstein-Barr Virus Latent Membrane 1 C-Terminal Activation Regions land 2[J].Journal of Virology,1999,73(7):5548-5555.
[9]Toshiyuki H.,Tomokazu Y.,Tzung-Shiahn S.et al.Association of latent membrane protein 1 and matrix metalloproteinase 9 with metastasis in nasopharyngeal carcinoma.Cancer,2000,89(4):715-723.
[10]Tomokazu Y,Hiroshi Sato,Mitsuru E et al.The expression of matrix metalloproteinase 9 is enhanced by Epstein-Barr virus latent membrane protein 1.Proc.Natl.Acad.USA 1998,95:3621-3626.
[11]Madigan MC,Kingsley EA,Cozzi PJ,et al.The role of extracellular matrix metalloproteinase inducer protein in prostate cancer progression[J].Cancer Immunol Immunother.2008,Feb 14[Epub ahead of print]
[12]Rollin J,Regina S,Vourch P,et al.Influence of MMP-2 and MMP-9promoter polymorphisms on gene expression and clinical outcome of non-small cell lung cancer.Lung cancer,2007 May;56(2):273-80.
[13]Ko HM,Kang JH,Jung B,et al.Critical role for matrix metalloproteinase-9 in platelet-activating factor-induced experimental tumor metastasis.Int J Cancer,2007 Mar 15;126(6):1277-83.
[14]Taras D,Blanc JF,Rullier A,et al.Pravastatin reduces lung metastasis of rat hepatocellular carcinoma via a coordinated decrease of MMP expression and activity.J Hepatol,2007 Jan;46(1):69-76.
[15]谢建武,方伟岗,李红梅等,金属蛋白酶MMP-9反义基因对高转移性肺巨细胞癌系的转染效应.中华病理学杂志,1999,28(4):210-211.
[16]牛桂英,方伟岗,郑杰等,TIMP-3基因转染抑制人肺癌细胞系浸润与转移的研究.中华病理学杂志,1998,27(6):421-424.
[17]Trulzsch B,Wood M.Applications of nucleic acid technology in the CNS.J Neurochem,2004;88(2):257-265.
[18]Finkel E.DNA cuts its teeth—as an enzyme.Science,1999;286:2441-2442.
[19]Kurreck J,Bieber B,Jahnel R,et al.Comparative study of DNA enzymes and ribozymes against the same full-length messenger RNA of the vanilloid receptor subtype I.J Biol Chem,2002;277(9):7099-7107.
[20]Breaker RR and Joyce GF.A DNA enzyme that cleaves RNA.Chem.Biol.1994;1(4):223-229.
[21]卢忠心,曹亚.脱氧核酶及其应用进展.国外医学分子生物学分册,2003;25(5):282-5.
[22]Santoro SW,Joyce GF,A general purpose RNA-cleaving DNAenzyme[J].Pro Nat Acad sci USA.1997,94:4262-4266.
[23]Unwalla H,Baner jea AC.Novel Mono-and-di-DNA-enzymes Targeted to Cleave TAT or TAT-REV RNA inhibit HIV-1 Gene Expression[J],Antiviral Res,2001,51(2):127-139.Application[J].Nucleosiddes Nuclweic Acid,2003,22(5):601-604.
[24]Wengle J,Vestev B.LNA and Alpha-L-LNA=TO wards Therapeutic Application[J].Nucleosiddes Nuclweic Acid,2003,22(5):601-604.
[25]Sioud M,Leirdal M,Dedsign of Nucles Resistant Protein Kinas Ca DNA Enzymes with potential therapelltic Application[J],.Mol Biol 2000,296(3):937-947.
[26]Warashine M,Kuwabara T,Nadamatsu Y,et al.Extremelly High and specipic Activity of DNA Enzymes in cells with philadelphiia Chromosome[J].Chem Biol,1999,6(4):237-250.
[27]于乐成,王升启,顾长海等,脱氧核酶再转基因肝癌细胞中对HCV-5非编码区的抑制活性[J],中华传染病杂志,2004,22(2):82-85.
[28]Yen L,Strittmatter SM,Kalb RG.Sequence-specific cleavage of Huntingtin mRNA by catalytic DNA[J].Ann Neurol,1999,46(3):366.
[29]Khachigian LM,Fahmy RG,Zhang G.et al.c-Jun regulates vascular smooth muscle cell growth and neointima formation after arterial injury.Inhibition by a novel DNA enzyme targeting c-Jun[J].J Biol Chem,2002,277(25):22985.
[30]Lowe HC,Chesterman CN,Khachigian LM.Catalytic antisense DNA molecules targeting Egr-1 inhibit neointima formation following permanent ligation of rat common carotid arteries[J].Thromb Haemost,2002,87(1):134.
[31]Dass CR,Choong PF,Khachigian LM.DNAzyme technology and cancer therapy:cleave and let die[J].Mol Cancer Ther.2008,7(2):243-51.
[32]Sasagawa T,Shimakage M,Nakamura M,et al.Epstein-Barr virus(EBV)genes expression in cervical intraepithelial neoplasia and invasive cervical cancer:a comparative study with human papilloma virus(HPV)infection.Hum Pathol.2000;31(3):318-326.
[33]Jen KY,Cheng J,Li J,et al.Mutational events in LMP1 gene of Epstein-Barr virus in salivary gland lymphoepithelial carcinomas.Int J Cancer.2003; 105(5):654-660.
[34]Kuo T,Hsueh C.Lymphoepithelioma-like salivary gland carcinoma in Taiwan:a clinicopathological study of nine cases demonstrating a strong association with Epstein-Barr virus.Histopathology.1997;31(1):75-82.
[35]Morbini P,Riboni R,Tomaselli S,et al.Eber- and LMP-l-expressing pulmonary lymphoepithelioma-like carcinoma in a Caucasian patient.Hum Pathol.2003;34(6):623-5.
[36]Lu ZX,Ma XQ,Yang LF,Wang ZL,DNAzymes targeted to EBV-encoded latent membrane protein-1 induce apoptosis and enhance radiosensitivity in nasopharyngeal carcinoma.Cancer Lett.2008 Mar 17.
[37]Rao JS,Gondi C,Chetty C,et al.Inhibition of invasion,angiogenesis,tumor growth,and metastasis by adenovirus-mediated transfer of antisense uPAR and MMP-9 in non-small cell lung cancer cells[J].Mol Cancer Ther.2005,4(9):1399-408.
[38]Tummalapalli P,Spomar D,Gondi CS,et al.RNAi-mediated abrogation of cathepsin B and MMP-9 gene expression in a malignant meningioma cell line leads to decreased tumor growth,invasion and angiogenesis[J].Int J Oncol.2007,31(5):1039-50.
[39]Tummalapalli P,Spomar D,Gondi CS,et al.RNAi-mediated abrogation of cathepsin B and MMP-9 gene expression in a malignant meningioma cell line leads to decreased tumor growth,invasion and angiogenesis[J].Int J Oncol.2007,31(5):1039-50.
[40]Nakamura T,Kuwai T,Kim JS,et al.Stromal metalloproteinase-9 is essential to angiogenesis and progressive growth of orthotopic human pancreatic cancer in parabiont nude mice[J].Neoplasia.2007,9(11):979-86.
[41]Zheng H,Takahashi H,Murai Y,et al.Expressions of MMP-2,MMP-9 and VEGF are closely linked to growth,invasion,metastasis and angiogenesis of gastric carcinoma[J].Anticancer Res.2006,26(5A):3579-83.
[42]藤博,辛丁,文莲姬等。10-23脱氧核酶靶向抑制喉癌eIF4E基因表达的实 验研究[J].临床耳鼻咽喉头颈外科杂志,2007,21(12):552-4.
[43]Zhang L,Gasper WJ,Stass SA,et al.Angiogenic inhibition mediated by a DNAzyme that taygets vascular endothelial growth factor receptor[J].Cancer Research,2002,62(19):5463-5469.
[44]Santoro SW,Joyce GF.Mechanism and utility of an RNA-cleaving DNA enzyme.Biochemistry.1998 Sep 22;37(38):13330-42.
[45]Cairns M J,Hopkins TM,Witherington C,et al.The influence of arm length asymmetry and base substitution on the activity of the 10-23 DNA enzyme.Antisense Nucleic Acid Drug Dev.2000;10(5):323-32.
[46]Pan WH,Devlin HF,Kelley C,et al.A selection system for identifying accessible sites in target RNAs.RNA.2001;7(4):610-21.
[47]Cairns M J,Hopkins TM,Witherington C,et al.Target site selection for an RNA-cleaving catalytic DNA.Nat Biotechnol.1999;17(5):480-6.
[48]Yen L,Strittmatter SM,Kalb RG.Sequence-specific cleavage of Huntingtin mRNA by catalytic DNA.Ann Neurol.1999;46(3):366-73.
[1] Ray M, Stetier-Stevenson W GGelatinase A activity directly modulatesmelanoma cell adhesion and spreading [J] EMBO J, 1995,14, (4):908-917
[2] Pereda MP,Hopfner U,Pagotto U,et al.Retinoic acid stinulates meningioma cell adhesion to the extracellumatrix and inhibits invasion [J],BrJ Cancer, 1999, 81(3):381-386
[3] Ueda M,Terai Y, Kumagai k, et al.Correlation between thymidine phosphorylase expression and invasion phenotype in cervical carcinoma cell [J], Int J Cancer, 2001, 91(6):778-782
[4] Masahito T, Jianguo G, Huan T, et al.PTEN gene and integrin signaling in cancer[J]. Journal of the Nation cancer Institute,1999,91(2):1820.
[5] Yang Y, Dang D, A takilit A,et al.Specific alpha vintegrin receptors modulate K1735 murine melanoma cell behavior [J],Bio Chem Biophys Res Commun, 2003, 308(4):814-819
[6] Clezardin P. Recent insights ioto the role of inegrins in cancer Metastasis[J]. Mol. Life. Sic, 1998, 54:541
[7] Grant MB, Caballero S,Kornberg L,et al. Focal Adhesion Kinase(FAK) Overexpression Modulates Retinal Angiogenesis [J] .Invest Ophthalmol Vis Sic, 2004,45:4463-4469.
[8] Gou XM, Chen XY, Arrand JR, et al. Effects of Epstein-Barr virus la tent membrane proteinl ( EBV-LMP1 ) on related factors of metastasis of nasopharyngeal carcinoma cell line CNE1 [J]. Ai Zheng, 2003, 22(5):481-485.
[9] Guishui Zhang, Crispin R. Dass, Eric Sumithran, et al. Effect of Deoxyribozymes Targeting c-Jun on Solid Tumor Growth and Angiogenesis in Rodents[J]. Journal of the National Cancer Institute, 2004, 96(9):683-696.
[10] Pasco S, Ramont L, Maquart FX, et al. Control of melanoma progression by various matriknes from basement membrane macromolecules [J]. Crit Rev Oncol Hematol, 2004,49:221-233.
[11] Eibl RH, Benoit M. Molecular resolution of cell adhesionforces[J].IEE Proc Nanobiotechnol.2004 Jun;151(3):128-32.
[12]Tamura M,Gu J,Matsumoto k,et al.Inhibition of cell migration,and focal adhesion by tumor suppressor PTEN[].Science,1998,280:1614.
[13]Pukac L,Huangpu J,Karnovsky MJ.Platelet-drived growth factor-BB,insulin-link growth factor-1,and phorbol ester activate different signaling pathway for stimulation of vascular smooth muscle cell migration[J].Exp.Cell.Res,1998,242:548.
[14]Sheppard D,Characterization of the integrin avβ6 as a fibronectin-binding protein[J].J.Bio.Chem,1992,267:5790.
[15]方新初,苏剑敏,周国非等。整合蛋白a5,β1亚基的过表达对肝癌细胞粘附与迁移的影响[J]。中国癌症杂志,2001,11(1):1-4。
[16]Taras D,Blanc JF,Rullier A,et al.Pravastatin reduces lung metastasis of rat hepatocellular carcinoma via a coordinated decrease of MMP expression and activity.J Hepatol,2007 Jan;46(1):69-76.
[17]Ko HM,Kang JH,Jung B,et al.Critical role for matrix metalloproteinase-9 in platelet-activating factor-induced experimental tumor metastasis.Int J Cancer,2007 Mar 15;126(6):1277-83.
[1]Santoro S W,Joyce GF.A general purpose RNA-cleaving DNA enzyme[J].Proc Natl Acad Sci USA,1997,94(9):4262
[2]Unwalla H,Baner jea AC.Novel Mono-and-di-DNA-enzymes Targeted to Cleave TAT or TAT-REV RNA inhibit HIV-1 Gene Expression[J],Antiviral Res,2001,51(2):127-139.Application[J].Nucleosiddes Nuclweic Acid,2003,22(5):601-604.
[3]Wengle J,Vestev B.LNA and Alpha-L-LNA=TO wards Therapeutic Application [J].Nucleosiddes Nuclweic Acid,2003,22(5):601-604.
[4]Sioud M,Leirdal M,Dedsign of Nucles Resistant Protein Kinas Ca DNA Enzymes with potential therapelltic Application[J],Mol Biol 2000,296(3):937-947.
[5]Warashine M,Kuwabara T,Nadamatsu Y,et al.Extremelly High and specipic Activity of DNA Enzymes in cells with philadelphiia Chromosome[J].Chem Biol,1999,6(4):237-250.
[6]于乐成,王升启,顾长海等,脱氧核酶再转基因肝癌细胞中对HCV-5非编码区的抑制活性[J],中华传染病杂志,2004,22(2):82-85.
[7]Yen L,Strittmatter SM,Kalb RG.Sequence-specific cleavage of Huntingtin mRNA by catalytic DNA[J].Ann Neurol,1999,46(3):366.
[8]Khachigian LM,Fahmy RG,Zhang G.et al.c-Jun regulates vascular smooth muscle cell growth and neointima formation after arterial injury.Inhibition by a novel DNA enzyme targeting c-Jun[J].J Biol Chem,2002,277(25):22985.
[9]Lowe HC,Chesterman CN,Khachigian LM.Catalytic antisense DNA molecules targeting Egr-1 inhibit neointima formation following permanent ligation of rat common carotid arteries[J].Thromb Haemost,2002,87(1):134.
[10]Emilsson GM,Breaker RR.Deoxyribozymes:new activities and new applications[J].Cell Mol Life Sci,2002,59(4):596
[11]Zuker M.Mfold web server for nucleic acid folding and hybridization prediction.Nucleic Acids Res.2003;31(13),3406-15.
[12]Zaborowska Z,Furste JP,Erdmann VA,et al.Sequence requirements in the catalytic core of the "10-23" DNA enzyme.J Biol Chem.2002;277(43):40617-22.
[13]Zaborowska Z,Schubert S,Kurreck J,et al.Deletion analysis in the catalytic region of the 10-23 DNA enzyme.FEBS Lett.2005;579(2):554-8.
[14]Cairns MJ,King A,Sun LQ.Optimisation of the 10-23 DNAzyme-substrate pairing interactions enhanced RNA cleavage activity at purine-cytosion target sites[J].Nucleic Acids Res,2003,31(11):2883-2889.
[15]Cairns MJ,Hopkins TM,Witherington C,et al.Target site selction for an RNA-cleaving catalytic DNA[J].Nat Biotechnol,1999,17(5):480-486
[16]Cairns MJ,Hopkins TM,witherington C,et al,The influence of arm length asymmetry and base substitution on the activity of the 10-23DNA enzyme[J].Antisense Nuleic Acid Drug Dev,2000,10(5):323-332
[17]Yen C,strittmater SM,kalb,RG.sequence-specific cleanage of Huntingon mRNA by Catalytic DNA[J].Ann Neurol,1999,46(3):366-373
[18]Sioud M,leirdal M.Design of nuclease resistant protein kinase k alpha DNA enzyme with potential therapeutic application[J].J Mol Bid,2000,296(3):937-947
[19]Asahine y,lto y,wu CH,et al.DNA ribonucleases that are active against intracellular hepatitis B viral RNA targets[J].Hepatology,1998,28(2):547-554.
[21]Vester B,Lundberg LB,sorensen MD,et al,lmprovd RNA cleavage by LNAzyme derivatives of DNAzymes[J].Biochen soc Trans,2004,32(p+1):37-40.
[22]Kubo T,Takamon K.Conjugate DNAzymes[J].Nucleic Acid Res,2003,3(Suppl):177-178
[23]Zaborowska 2,Furste JP,Erdmann VA.etal.Sequence requirements in the catalytic core of the "10-23"DNA enzyme[J].J Biol chem,2002,277(43):40617-40622
[24]Abdelqang A,wood M,Beeson D.Hairpin DNAzymes:a new tool for efficient cellular gene silencing[J].Gene Med,2007,Aug;9(8):727-738
[25] Jakobsen MR,Haasnoot J, Wenqel J, et al.Eefficient inhibition of HIV-1 expression by LNA modiffied antisense digonucleotides and DNAzymes targeted to functionlly selected binding sites, Retrovirology, 2007, Apr, 26(4):29
[26] Zhang L,Gasper WJ, stass SA, et al .Angiogenic inhibition mediated by a DNAzyme that targets Vascular endothelial growth factor receptor 2[J], Cancer Research, 2002, 62(19):5463-5469.
[27] Liu C, cheng R,Sun LQ, et al.Suppression of platelettype 12-lipoxygenase activity in human erythroleuke miacells by an KNA-cleaving DNAzyme[J], Biochem Biophys Res com, 2001.284(4):1077
[28] Wu Y, Yu L,Mcmahon R,et al.Inhibition of bcr-abl oncogen expression by novel dexyribozymes(DNAzymes)[J].Humei Gene Ther,1999,10(17):2847-2857.
[29] Liang Z, Wei S,Guan J, et al.DNAzyme mediated cleavage of surviving mRNA and inhibition of the growth of PANC-1 cell[J]. J Gastroenterol Hepatol, 2005, 20(10):1595