应用RNA干扰技术建立MMP-9基因沉默胃癌细胞克隆
|
摘要
恶性肿瘤的主要特征是浸润和转移,细胞外基质(ECM)和基底膜的降解为关键环节。基质金属蛋白酶-9(MMP-9)作为蛋白水解酶的一种,能降解基底膜和细胞外基质成分,从而促进肿瘤的浸润、转移及间质血管新生。胃癌组织中MMP-9检测结果与临床病理资料对比的研究表明,该酶不仅与胃癌的分化程度、分期、病理类型、转移和预后有关,然而MMP-9基因在转录和翻译水平的抑制作用尤其是RNA干扰对胃癌的影响尚未明了。
RNA干扰是由dsRNA介导的同源mRNA降解,从而引起相应基因沉默的一种广泛存在于酵母、真菌、植物、动物体内古老的生物防御机制。1998年后,许多研究小组对RNAi的机制和应用进行了大量的研究。在应用方面主要是利用RNAi技术对线虫、果蝇、植物、动物及病毒的某些基因进行干扰,从而达到基因敲除或抑制病毒复制的目的。
本实验目的是应用质粒载体将基质金属蛋白酶-9(MMP-9)特异性siRNA通过脂质体转染胃癌细胞,构建了低分化胃腺癌细胞BGC823的MMP-9基因沉默细胞模型;应用免疫荧光、western blot、明胶酶谱分析等方法验证RNA干扰所得到的基因沉默胃癌细胞株。
本实验应用分子克隆技术结合RNA干扰技术在转录和翻译水平对特定基因MMP-9进行抑制,从而对该基因在胃癌浸润、转移中的作用及机制进行探讨。实验关键在于获得较高干扰效果并筛选获得成功转染质粒的细胞克隆。这将为后续的体内外胃癌模型建立提供基础。该RNA干扰模型建立,从分子水平探讨胃癌细胞浸润与基质金属蛋白酶9的关系,在胃癌基因水平研究做了初步探索,结果可能对胃癌的早期诊断、预后判断和基因治疗提供参考。
The major characteristics of malignant tumour cells are invasion and metastasis, of which proteolytic degradation of the extra cellular matrix, including basement membrane by matrixmetalloproteinases (MMPs) is one of the essential events. As a kind of matrixmetallo-proteinases, matrixmetalloproteinase-9(MMP-9) can degradate extra-cellular matrix and basement membrane, which would facilitate the invasion, metastasis and interstitial angiogenesis. MMP-9 is expressed predominantly by tumour cells in various cancers. Immunohistochemical studies have shown that the expression of MMP-9 correlates with tumor invasion,metastasis and prognosis in both colorectal adenocarcinoma and esophageal squamous cell carcinoma. But it is not certain what will happen when MMP-9 is inhibitted with RNAi.
Over the last years, RNA interference(RNAi) has been recognized as a major mechanism of post-transcriptional gene silence in the nematode Caenorhabditis elegans and the fruit fly Drosophila, as well as in plants. This phenomenon is based on double-stranded RNA(dsRNA) that triggers the silence of gene expression in a sequence-specific manner. Now, there are many study on the mechanism, while has become a routine tool for transient knockdown of gene expression in a wide range of organisms. In our experiment, we constructed a modal with MMP-9 gene
knockout by RNAi in poorly differentiated gastric adenocarcinoma cell BGC823. In the process of construction, we cloned a stable RNAi system with liposome transfection. Then applied immunofluorescence technique , gelatin zymography and western blot to detect the expression of MMP-9 in BGC823.
The experiment applies molecular cloning and RNA infection techniques to inhibit MMP-9 gene in level of transcription and translation, for researching the role and mechanism of the invasion and metastasis in gastric cancer cell. Experimental key is to obtain a higher interference effects gastric cancer cell line that transfected plasmid successfully. The experiment is the basis of the experimental model in vitro. The resulds would show the relationship between gastric cancer cell invasion and MMP-9, provide evidences for early diagnosis, gene therapy, and judging the prognosis in gastric cancer.
RNA干扰是由dsRNA介导的同源mRNA降解,从而引起相应基因沉默的一种广泛存在于酵母、真菌、植物、动物体内古老的生物防御机制。1998年后,许多研究小组对RNAi的机制和应用进行了大量的研究。在应用方面主要是利用RNAi技术对线虫、果蝇、植物、动物及病毒的某些基因进行干扰,从而达到基因敲除或抑制病毒复制的目的。
本实验目的是应用质粒载体将基质金属蛋白酶-9(MMP-9)特异性siRNA通过脂质体转染胃癌细胞,构建了低分化胃腺癌细胞BGC823的MMP-9基因沉默细胞模型;应用免疫荧光、western blot、明胶酶谱分析等方法验证RNA干扰所得到的基因沉默胃癌细胞株。
本实验应用分子克隆技术结合RNA干扰技术在转录和翻译水平对特定基因MMP-9进行抑制,从而对该基因在胃癌浸润、转移中的作用及机制进行探讨。实验关键在于获得较高干扰效果并筛选获得成功转染质粒的细胞克隆。这将为后续的体内外胃癌模型建立提供基础。该RNA干扰模型建立,从分子水平探讨胃癌细胞浸润与基质金属蛋白酶9的关系,在胃癌基因水平研究做了初步探索,结果可能对胃癌的早期诊断、预后判断和基因治疗提供参考。
The major characteristics of malignant tumour cells are invasion and metastasis, of which proteolytic degradation of the extra cellular matrix, including basement membrane by matrixmetalloproteinases (MMPs) is one of the essential events. As a kind of matrixmetallo-proteinases, matrixmetalloproteinase-9(MMP-9) can degradate extra-cellular matrix and basement membrane, which would facilitate the invasion, metastasis and interstitial angiogenesis. MMP-9 is expressed predominantly by tumour cells in various cancers. Immunohistochemical studies have shown that the expression of MMP-9 correlates with tumor invasion,metastasis and prognosis in both colorectal adenocarcinoma and esophageal squamous cell carcinoma. But it is not certain what will happen when MMP-9 is inhibitted with RNAi.
Over the last years, RNA interference(RNAi) has been recognized as a major mechanism of post-transcriptional gene silence in the nematode Caenorhabditis elegans and the fruit fly Drosophila, as well as in plants. This phenomenon is based on double-stranded RNA(dsRNA) that triggers the silence of gene expression in a sequence-specific manner. Now, there are many study on the mechanism, while has become a routine tool for transient knockdown of gene expression in a wide range of organisms. In our experiment, we constructed a modal with MMP-9 gene
knockout by RNAi in poorly differentiated gastric adenocarcinoma cell BGC823. In the process of construction, we cloned a stable RNAi system with liposome transfection. Then applied immunofluorescence technique , gelatin zymography and western blot to detect the expression of MMP-9 in BGC823.
The experiment applies molecular cloning and RNA infection techniques to inhibit MMP-9 gene in level of transcription and translation, for researching the role and mechanism of the invasion and metastasis in gastric cancer cell. Experimental key is to obtain a higher interference effects gastric cancer cell line that transfected plasmid successfully. The experiment is the basis of the experimental model in vitro. The resulds would show the relationship between gastric cancer cell invasion and MMP-9, provide evidences for early diagnosis, gene therapy, and judging the prognosis in gastric cancer.
引文
[1] Huang Hai-li,Wang Meng-wei.Advances in the Molecular Mechanisms of Tumor Infiltration and Metastasis.Letters In Biotechnology,2006;1:84-87.
[2] Matvey E,Lukashev,Zena Werb,et al.ECM signalling: orchestrating cell behaviour and misbehaviour.Trends in Cell Biology, 1998;8:437-441.
[3] The role of gelatinases in colorectal cancer progression and metastasis. Biochimica et Biophysica Acta (BBA) - Reviews on Cancer.2004;1705:69-89.
[4] Kenny HA,Kaur S,Coussens LM,et,al.The initial steps of ovarian cancer cell metastasis are mediated by MMP-2 cleavage of vitronectin and fibronectin.J Clin Invest.2008;118:1367-1379.
[5] Visse R,Nagase H.Matrix Metalloproteinases and Tissue Inhibitors of Metalloroteinases:Structure,Function,and Biochemistry.Circulation Research,2003;92:827-839.
[6] Brew K,Dinakarpandian D,Nagase H.Tissue inhibitors of metalloproteinases:evolution,structure and function.Biochim Biophys Acta,2000;1477:267-283.
[7] Yoon SO,Park SJ,Yun CH,et al.Roles of Matrix Metalloproteinases in Tumor Metastasis and Angiogenesis.Journal of Biochemistry and Molecular Biology,2003;36:128-137.
[8] Bogenrieder T,Herlyn M.Axis of evil:molecular mechanisms of cancer metastasis,Oneogene,2003;22:6524-6536.
[9] Woessner JF Jr.Matrix metalloproteinases and their inhibitors in connective tissue remodeling.FASEB J,1991;5:2145-2154.
[10] Connor CMO,Fitz Gerald MX.Matrix metalloproteinases and lung disease.Thorax,1994;49:602.
[11] Nagase H,Woessner JF Jr.Matrix metalloproteinases.J Biol Chem.1999;2742:1491-1494.
[12] Krejci P,Masri B,Fontaine V,et al.Interaction of fibroblast growth factor and C-natriuretic peptide signaling in regulation of chondrocyte proliferation and extracellular matrix homeostasis.J Cell Sci. 2005;118:5089-5100.
[13] Birkedal Hansen H,Moore WG I,Bodden,et a1.Matrixmetalloproteinases.Crit Rev Oral Biol Med,1993;4:97.
[14] LI Jing,ZHAO Tianfu,DUAN Enkui.Matrix metalloproteinases (MMPs) and trophoblast invasion.science report,2005;12:1169-1173.
[15] Rhee JS,Coussens LM.RECKing MMP function:implications for cancer development.Trends Cell Biol.2002;12:209-211.
[16] Huang H, Wang M. Advances in the Molecular Mechanisms of Tumor Infiltration and Metastasis.Letters in biotechnology,2006 ;7: 54-56.
[17] Liotta LA, Steeg PS, Stetler-Stevenson WG.Cancer metastasis and angiogenesis:an imbalance of positive and negative regulation.Cell,1991;64:327-336.
[18] Crawford, H.c.Martrisian,et al.L.M Tumor stromal expression of matrixmetall oproteinases their role in tumor progression.Invasion metastasis,l994;14:234-245.
[19] Stetler-stervenson,W.G Aznavoortan,S Liotta,LA,et al.Tumor cell interaction with the extra cellular matrix during invasinon and metastasis.Annu Rev Cell Biol, 1993;9:541-573.
[20] Liotta LA,Tryggvason K,Garbisa S,et al.Metastatic potential correlates with enzymatic degradation of basement membrane collagen.Nature,1980;284:67-68.
[21] DaviesB,MilesDW,Happerfield LC,et al.Levels of type IV collagenases in benign and malignant breast disease.Br J Cancer Res,1993;67:1126-1131.
[22] Davies B,Waxman J,Wasan H,et al.Levels of Matrixmetalloproteinases in bladder cancer correlate with tumor grade invasion.Cancer Res,1993;53:5365-5369.
[23] Chambers AF,Matrisian L,et al.Changing views of the role of matrMetall opro- teinases in metastasis.Natl Cancer Inst.1997;89:1260-1270.
[24] Kahari VM,Saraarial ho-KereU,et al,matrixmetalloproteinases and the Inhibitors in tumor growth invasion.Ann Med,1999;31:34-45.
[25] Ray JM,Stetler Stevenson WG,et al.Gelatinase A activity directly modulate Mela- noma cell adhesion spreading.EMBO,1995;14:908-917.
[26] Kleiner DE,Stetler Stevenson WG.Matrixmetalloproteinases and metastasis. Can Che Pha,1999;43:342-351.
[27] Sauter W,Rosenberger A,Beckmann L,et al.Matrix Metalloproteinase 1(MMP1) Is Associated with Early-Onset Lung Cancer.Cancer Epidemiol Biomarkers Prev.2008;17:1127-1135.
[28] Alfano RW,Leppla SH,Liu S,et al.Cytotoxicity of the matrix metalloproteinase-activated anthrax lethal toxin is dependent on gelatinase expression and B-RAF status in human melanoma cells.Mol Cancer Ther.2008;7:1218-1226.
[29] De Cicco C,Ravasi L,Zorzino L,et al.Circulating levels of VCAM and MMP-2 may help identify patients with more aggressive prostate cancer.Curr Cancer Drug Targets.2008;8:199-206.
[30] Bendrik C,Robertson J,Gauldie J,et al.Gene transfer of matrix metalloproteinase-9 induces tumor regression of breast cancer in vivo.Cancer Res.2008;68:3405-1342.
[31] Lakka SS,Gondi CS,Yanamandra N,et al.Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis.Oncogene,2004;23:4681-4689.
[32] Ito H,Duxbury M,Benoit E,et al.Fibronectin-induced COX-2 mediates MMP-2 expression and invasiveness of rhabdomyosarcoma.Biochem Biophys Res Commun.2004;318:594-600
[33] Marin-Casta?o ME, Csaky KG, Cousins SW.Nonlethal oxidant injury to human retinal pigment epithelium cells causes cell membrane blebbing but decreased MMP-2 activity.Invest Ophthalmol Vis Sci. 2005 ;46:3331-3340.
[34] Karagiannis ED, Popel AS. Distinct modes of collagen type I proteolysis bymatrix metalloproteinase (MMP) 2 and membrane type I MMP during the migration of a tip endothelial cell: insights from a computational model.J Theor Biol. 2006;238:124-145.
[35] Bai YL, Huang HC, Li JZ, Zhao YY, Wang HY.High glucose regulates the production of MMP-9 in podocyte through ERK1/2 signal pathway.Zhonghua Yi Xue Za Zhi,2005;85:1451-1455.
[36] Azkur AK, Kim B, Suvas S, Lee Y, Kumaraguru U, Rouse BT. Blocking mouse MMP-9 production in tumor cells and mouse cornea by short hairpin (sh) RNA encoding plasmids.Oligonucleotides. 2005;15:72-84
[37] Giraudo E, Inoue M, Hanahan D.An amino-bisphosphonate targets MMP-9-expressing macrophages and angiogenesis to impair cervical carcinogenesis.J Clin Invest.2004;114:623-633.
[38] Topp SA, Upadhya GA, Strasberg SM. Cold preservation of isolated sinusoidal endothelial cells in MMP 9 knockout mice: effect on morphology and platelet adhesion.Liver Transpl. 2004;10:1041-1048.
[39] Liotta LA,Tryggvason K,Garbisa S,et a1.Metastatic potential correlates with enzymatic degradation of basement membrane collagen.Nature,1980;284:67
[40] Albini A,Melchiori A,Santi L,et a1.Tumor cell invasion inhibited by TIMP-2. Natl Cancer Inst,1991;83:775.
[41] Kawamata H,Kameyama S,Kawai K,et a1.Marked acceleration of the metastatic phenotype of a rat bladder carcinoma cell line by the expression of human gelatinase A.Int J Cancer,1995;63:568.
[42] Kawamata H,Kawai K,Kameyama S,et al. Over-expression of tissue inhibitor of matrix metalloproteinases (TIMP1 and TIMP2) suppresses extravasation of pulmonary metastasis of a rat bladder carcinoma.Int J Cancer,1995;63:680.
[43] Ray JM,Stetler Stevenson WG.The role of matrix matalloproteases and their inhibitors in tumour invasion,metastasis and angiogenesis.Eur Respir J,1994;7:2062.
[44] Yoshikawa T,Tsuburaya A,Kobayashi O,et al.Intratumoral concentrations oftissue inhibitor of matrix metalloproteinase Iin patients with gastric carcinoma a new biomartker for invasion and its impact on survival.Cancer,2001;91:1739.
[45] Garzetti G,Andrea C,Guendalina L,et a1.Expression of vascular endothelial growth factor related to 72-Kiloalton metalloproteinase immunostaining patients with seriousovarian tumor.Cancer,1999;85:2219-2225.
[46] Schwartz GK,Warg H,Lampen N,et a1.Defining the invasive phenol type of proximal gastric cancer cells.Cancer,1994;73:222-227.
[47] Li nan,Caipu Xu,Weiwen Lu,et al.Effect of tissue inhibitor of matrix metalloproteinase-2 spontanous invasion of gastric carcinoma cell lines in vitro. China Oncology,2000;10:163-165.
[48] Li nan,Caipu Xu,Weiwen Lu,et al.Effect of tissue inhibitor of matrix metalloproteinase-2 spontanous invasion of gastric carcinoma cell lines in vitro and in vivo.China Oncology .Chinese Journal of Pathology,1998;27:362-365.
[49] Inoue T,Yashiro M,Nishimra S.Matrix metalloproteinases1 expression is a prognostic factor for Patients with advanced gastric cancer.In J Mol-Med, 1999;4:73-77.
[50] Li L,Zhang sheng,Lin hua,et al.Relationship of Expression Unbalance of Matrix Metalloproteinase and Tissue Inhibitor of Metalloproteinase to Invasiveness and Metastasis in Gastric Carcinomas. Chi Jou of Can,2002;21:305-310.
[51] Yoshikawa T,Tsuburaya A,KobayashiO,et al.Prognostic value of tissue inhibitor of matrix metalloproteinase-I in plasma of patients with gastric cancer.Cancer Lett,2000;151:81-86.
[52] Zhang S,Li L,Lin J, et al.Imbalance between expression of matrix metalloproteinase-9 and tissue inhibitor of metaloproteinase-1 in invasiveness and metastasis of human gastric carcinoma.Wor J Gas,2003;9:899-904.
[53] Yutaka Takahashi,Yasuhiko Kitadai,Lee M.et al.Multiparametric in situmRNA Hybridization Analysis of Gastric Biopsies Predicts Lymph Node Metastasis in Patients with Gastric Carcinoma.Cancer Res,2002;93:1258-1265.
[54] Xiaoping Miao, Chunyuan Yu,Wen Tan,et al.A Functional Polymorphism in the Matrix Metalloproteinase-2 Gene Promoter Is Associated with Risk of Development but not Metastasis of Gastric Cardia Adenocarcinoma. Cancer Research,2003;63:3987-3990.
[55] Matsumura S,Oue N,Nakayama H,et al.A single nucleotide polymorphism in the MMP-9 promoter affects tumor progression and invasive phenotype of gastric cancer.Can Res Clin Onco,2005;131:19-25.
[56] Koyama S.Enhanced cell surface expression of inhibitors,and tumor-induced host response matrix metalloproteinases and their in progression of human gastric carcinoma.Dig Dis Sci,2004;49:1621-1630.
[57] Murray GI,Duncan ME,Arbuckle E,Melvin WT,Fothergill JE.Matrix metalloproteinases and their inhibitors in gastric cancer.Gut,1998;43:791-797.
[58] Tsuchiya Y,Sato H,Endo Y,et al.Tissue inhibitor of metalloproteinase 1 is a negative regulator of the metastatic ability of a human gastric cancer cell line, KKLS, in the chick embryo.Cancer Res,1993;53:1397-402.
[59] Nomura H,Fujimoto N,Seiki M,et al.Enhanced production of matrix metalloproteinases and activation of matrix metalloproteinase 2 (gelatinase A) in human gastric carcinomas.Int J Cancer,1996;69:9-16.
[60] Lisa M.C,Barbara Fingleton,and Lynn M.Matrisian Matrix Metalloproteinase Inhibitors and Cancer:Trialsand Tribulations.Science,Mar,2002;295:2387-2392.
[61] Sang-Oh Yoon,Soo-Jin Park,Chang-Hyun Yun,eta1.Roles of in Tumor Matastasis and Angiogenesis.Journal of Biochemistry and Molecular Biology,2003;36:128-137.
[62] Price A,SW Q,Morris D,et al.Marked inhibition of tumor growth in amalignant glioma tumor model by a novel synthetic matrixmetalloproteinase inhibitorAG3340.Clin Cancer Res,1999;5:845-854.
[63] Knox JD,Bretton L,Lynch et al.Synthetic matrix metalloproteinase inhibitor,BB-94,inhibits the invasion of neoplastic human prostate cells in a mouse model.Prostate,1998;35:248-254.
[64] Anderson IC,Shipp MA,Docherty AJ,et al.Combination therapy including a gelatinase inhibitor and cytotoxic agent reduces local invasion and metastasis of murine Lewis lung carcinoma.Cancer Res,1996;56:715-718.
[65] Nicoletti V,Lucchini M,D'Incalci R,et a1.Comparison of paclitaxel and docetaxel activity on human ovarian carcinoma xenografts.European Journal of Cancer, 1994;30:691-696.
[66] Godehard Friedel,Ugo Pastorino,Robert J,et al.Results of lung metastasectomy from breast cancer: prognostic criteria on the basis of 467 cases of the international registry of lung metastases. European Journal of Cardio-Thoracic Surgery,2002;22:335-344.
[67] Bodurtha A,Eisenhauer E,Steward W,et al.Phase I-II study of marimastat(BB-2516) in patients with metastatic melanoma.Proc Am Soc Clin Oncol,1997;16:493.
[68] Parsons SL,Watson SA,Griffin NR,et a1.An open phase I/II study of the oral matrix metalloproteinase inhibitor marimastat in patients with inoperable gastric cancer.Ann Oncol,1996;47:47.
[69] Parano JA,Bernardo P,Stephenson P,et al.Randomized phase III trial of marimastat versus placebo in patients with metastatic breast cancer who have responding or stable disease after first-line chemotherapy.Clin Oncol,2004;22:4683-4690.
[70] Nemunaitis J,Poole C,Primrose J,et al.Combined analysis of studies of the effects of the matrix metalloproteinase inhibitor marimastat on serum tumor markers in advanced cancer:Selection of abiologically active and tolerable dose for longer-term studies.ClinCancerRes,1998;4:1101-1109.
[71] Rosemurgy A,Buckels J,Charnley R.Arandomized study comparing marimastat to gemcitabine as first line therapy in patients withnon-resectable pancreatic cancer.Proc Am Soc Clin Oncol,1999;18:261.
[72] Gradishar W,Sparano J,Cobleigh M,et al,A phase I study of marimastat in combination with doxorubicin and cyclophosphamide in patients with metastatic breast cancer.Proc Am Soc Clin Onco1,1998;17:144.
[73] Wildingq Small E,Rippleq et al,Phase I study of AG3340,a matrix metalloproteinase inhibitor incombination with mitoxantronel prednisonein patients having advanced prostate cancer.Ann Oncol,1998;9:74.
[74] Ikeda T,Murakami K,Hayakawa Y,et al.Anti-invasive activity of synthetic serine protease inhibitors and its combined effect with a matrix metalloproteinase inhibitor.Anti cancer Res,1998;18:4259-4265.
[75] Fire A,Xu S,Montgomery MK,et al.Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans.Nature,1998;391:806-811.
[76] Cogoni C,Macino G.Posttranscriptional gene silencing in Neurospora by a RecQ DNA helicase.Science,1999;286:2342-2344.
[77] Hannon GJ.RNA interference. Nature ,2002;418:244-251.
[78] Mario Stevenson.Dissecting HIV-1 through RNA interference.Nat Rev Immunol,2003;3:851-858.
[79] Elbashir SM,et al. Duplexes of 21 nucleotide RNAs mediate RNA interference in culture mammalian cells.Nature,2001;411:494-498.
[80] Elbashir SM,et al. RNA interference is mediated by 21-and 22-nucleotide RNAs .Genes Dev,2001;15:188-200.
[81] Verma NK,et al.RNA-mediated gene silencing:mecha-nisms and its therapeutic applications.J Clin Pharm T-her.2004,29:395-404.
[82] Angela Reynolds,et al.Rational siRNA design for RNA in-terference. Nature biotechnology,2004;22:326-330.
[83] Phillip D,et al.Ancient pathways programmed by small RNA.Science,2002;296:1265-1269.
[84] Ronald HAP. RNA silencing:the genome,s immune system.Science,2002;296:1263-1265.
[85] Michael B,et al. Genome-wide RNAi analysis of growth and viability in drosophila cells. Science,2004;303:832-835
[86] Verma NK,et al. RNA-mediated gene silencing:mechanisms and its therapeutic applications.J Clin Pharm T-her.2004;29:395 - 404.
[87] McCaffrey AP,et al. RNA interference in adult mice.Nature,2002;418:38-39.
[88] Kronke J,et al. Alternative approaches for efficient in-hibition of hepatitis C virus RNA replication by small in-terfering RNAs. J Virol,2004;78:3436-3446.
[89] Takigawa Y,et al.Suppression of hepatitis C virus replicon by RNA interference directed against the NS3 and NS5B regions of the viral genome.Microbiol Immunol,2004;48:591-598.
[90] George JA, Gene therapy progress and prospects:adenoviral vectors,Gene Therapy,2003;6:12-16.
[91] Liotta LA,Kohn EC,Cancer's deadly signature.Nat Genet,2003;33:49-54
[92] Li yingping,Zhang ying,Shao guoqing,A New Technology to Delete Gene-RNAi,Bio Bull,2006;3:27-30.
[93] Marjori M,et al.RNA:guiding gene silencing. Science,2001;293:1080-1083.
[94] Verma NK,et al.RNA-mediated gene silencing:mechanisms and its therapeutic applications.J Clin Pharm Ther.2004;29:395-404.
[95] Jun Yuan A, Charyl M,Dutton B,Sean P,et al.RNAi mediated MMP-1 silencing inhibits human chondrosarcoma invasion.Jou of Orth Res,2005;23:1467-1474.
[96] Brummelkamp TR,Bernards R,Agami R.Stable suppression of tumor igenicity by virus-mediated RNA interference.Cancer Cell,2002;2:243-247.
[97] Fleming JB,Shen GL,Holloway SE,et al.Molecular consequences of silencing mutant K-ras in pancreatic cancer cells:justification for K-ras-directed therapy.Mol Cancer Res,2005;3:413-423.
[98] Wang YH,Liu S,Zhang G,et al.Knockdown of c-Myc expression by RNAi inhibits MCF-7 breast tumor cells growth in vitro and in vivo.Breast CancerRes,2005;7:220-228.
[99] Takada Y,Sethi G,Sung B,et al.Flavopiridol suppresses tumor necrosis factor-induced activation of activator protein-1,c-Jun N-terminal kinase,p38 mitogen-activated protein kinase (MAPK),p44/p42 MAPK,and Akt,inhibits expression of antiapoptotic gene products,and enhances apoptosis through cytochrome c release and caspase activation in human myeloid cells.Mol Pharmacol.2008;73:1549-1557.
[100] Won KA,Reed SI.Activation of cyclin E/CDK2 is coupled to site-specific autophosphorylation and ubiquitin-dependent degradation of cyclin E.EMBO J.1996;15:4182-4193.
[101] Terano T,Tanaka T,Tamura Y,et al.Eicosapentaenoic acid and docosahexaenoic acid inhibit vascular smooth muscle cell proliferation by inhibiting phosphorylation of Cdk2-cyclinE complex.Biochem Biophys Res Commun.1999;254:502-506.
[102] Li K,Lin S,Brunicardi FC,et al.Use of RNA interference to target cyclin E over expressing hepatocellular carcinoma.Cancer Res,2003,63:3593-3597.
[103] Martin-Lluesma S, Stucke VM,Nigg EA.Role of Hec1 in spin-dle checkpoint signaling and kinetochore recruitment of Mad1/Mad2.Science,2002;297:2267-2270.
[104] Chen Z,Varney ML,Backora MW,et al.Down-regulation of vascular endothelial cell growth factor-C expression using small interfering RNA vectors in mammary tumors inhibits tumor lymphangiogenesis and spontaneous metastasis and enhances survival.Cancer Res,2005;65:9004-9011.
[105] Shen J,Samul R,Silva RL,et al.Suppression of ocular neovascularization with siRNA targeting VEGF receptor 1.Gene Ther,2006;13:225-234.
[106] Amantana A,London CA, Iversen PL,et al.X-linked inhibitor of apoptosis protein inhibition induces apoptosis and enhances chemotherapy sensitivity in human prostate cancer cells. Mol Cancer Ther,2004;3:699-707.
[107] Stierle V,Laigle A,Jolles B.Modulation of MDR1 gene expression inmultidrug resistant MCF7 cells by low concentrations of small interfering RNAs.Biochem Pharmacol,2005;70:1424-1430.
[108] .Pichler A,Zelcer N,Prior JL.In vivo RNA interference-mediated ablation of MDR1 P-glycoprotein.Clin Cancer Res,2005;11:4487-4494.
[109] Kami k,Doi R,Koizumi M,et al.Downregulation of surviving by siRNA diminishes radioresistance of pancreatic cancer cells.Surgery,2005;138:299-305.
[110] Tuschl T,Zamore PD,Lehmann R,et al.Targeted mRNA degradation by double-stranded RNA in vitro.Genes Dev,1999;13:3191-3197.
[111] Bernstein E,Caudy AA,Hammond SM,Hannon GJ.Role for a bidentate ribonuclease in the initiation step of RNA interference.Nature,2001;409:363-366.
[112] Zamore PD,Tuschl T,Sharp PA,Bartel DP.RNAi:double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals.Cell,2000;101:25-33.
[113] Paddison PJ,Caudy AA,Bemstein E,et a1.Short hairpin RNAszz (shRNAs)induce sequence-specific silencing in mammalian cells.Genes 2002;16:948-958.
[114] Miyagishi M,Taira K.U6 promoter-driven siRNAs with four uridine 3’overhangs efficiently suppress,targeted gene expression in mammalian cells.Nat Biotech,2002;19:497-500.
[115] Brummelkamp TR,Bernads R,Agami R.A system for stable expression of short interfering RNAs in mammalian cells.Science,2002;296:550-553
[116] Sachidanandam R.RNAi:design and analysis.Curr Protoc Bioinformatics.2004;12:Unit 12.3
[117] Kuittinen O,Soini Y,Turpeenniemi-Hujanen T.Diverse role of MMP-2 and MMP-9 in the clinicopathological behavior of Hodgkin's lymphoma.Eur J Haematol.2002;69:205-212.
[118] von Lüttichau I,Djafarzadeh R,Henger A,et al. Identification of a signal transduction pathway that regulates MMP-9 mRNA expression inglomerular injury.Biol Chem.2002;383:1271-1275.
[119] Lahat N,Shapiro S,Froom P,et al.Inorganic lead enhances cytokine-induced elevation of matrix metalloproteinase MMP-9 expression in glial cells.J Neuroimmunol.2002;132:123-128.
[120] Fang J,Shing Y,Wiederschain D,et al.Matrix metalloproteinase-2 is required for the angiogenic phenotype in a tumormodel.Proc Natl Acad Sci USA,2000;97:3884.
[121] Curran S,Murray GI.Matrix metalloproteinases in tumour invasion and metastasis.J Pathol,1999;189:300-308.
[122] Coussens LM,Fingleton B,Matrisian LM.Matrix metalloproteinases,inhibitors and cancer:trials and tribulations.Science,2002;295:2387-2391.
[123] Itoh T,Tanioka M,Yoshida H,et al.Reduced angiogenesis and tumor progression in gelatinase Adeficient mice.CancerRes,1998;58:1048-1051.
[124] NelsonAR,Fingleton B,Rothenberg ML,MatrisianLM.Matrix metal oproteinases:biologicactivity and clinicali mplications.J Clio Oncol, 2000;18:1135-1149.
[125] Schmalfeldt B,Prechtel D,Harting K,et al.Increased expression of matrixmetalloproteinases(MMP)-2,MMP-9,andtheurokinase-type plasminogen activator is associated with progression from benign to advancedovarian cancer.Clin Cancer Res,2001;7:2396-404.
[126] Yanagisawa N, Geironson L, Waleed AS, et al. Expression of matrix metalloprotease-2,-7,and -9 on human colon, liver and bile duct cell lines by enteric and gastric Helicobacter species.FEMS.Immunol Med Microbiol,2005;44:197-204.
[127] Sier CF,Kabben FJ,Ganesh S,et al.Tissue levels of matrix metalloproteinases MMP-2 and MMP-9 are related to the overall survival of patients with gastric carcinoma.Br J Cancer,1996;74:413-417.
[128] Wu ZY, Li JH, Zhan WH, et al. Lymph node micrometastasis and its correlation with MMP-2 expression in gastric carcinoma.World JGastroenterol, 2006;12:2941-2944.
[129] Sham S K,Mohammad TM.Regulation of angiogenesis and invasion by human pituitary tumor transforming gene(PTTG) through increased expression and secretion of matrix metalloproteinase-2 (MMP-2).Molecular Cancer,2006;5:61.
[130] Nomura H,Sato H,Seiki M,et al.Expression of membrane type matrix metalloproteinase in human gastric carcinomas.Cancer Res,1995;55:3263-3266.
[131] Nomura H,Fujimoto N,Seiki M,et al.Enhanced production of matrix metalloproteinases and activation of matrix metalloproteinase 2 (gelatinaseA)in human gastric carcinomas.Int J Cancer,1996;69:9-16.
[132] Kubben FJ,Sier CF,van Duijn W,et al.Matrix metalloproteinase-2 is a consistent prognostic factor in gastric cancer.Br J Cancer,2006;10,94:1035-1040.
[133] Chang C,Werb Z. The many faces of metalloproteases: cell growth, invasion, angiogenesis and metastasis.Trends Cell Biol,2001;11:S37-S43.
[134] Tuschl T. Expanding small RNA interference. Nat Biotechnol, 2002; 20:446-448.
[135] McManus MT, Sharp PA. Gene silencing in mammals by small interfering RNAs. Nat Rev Genet, 2002;3:737-747.
[136] Wang J, Tekle E, Oubrahim H, et al. Stable and controllable RNA inteference:investigating the physiological function of glutathionylated actin. Proc Natl Acad Sci USA, 2003;100:5103-5106.
[137] Zhou bo,Yang wei,Jianguo Ji,et al.Differential Protein Expression Induced by Transient Transfection of Metallothionein-3 Gene in SH-SY5Y Neuroblastoma Cell LineDifferential Protein Expression Induced by Transient Transfection of Metallothionein-3 Gene in SH-SY5Y Neuroblastoma Cell Line. ACTA BIOCHIMICA ET BIOPHYSICA SINICA. 2003;6:522-528.
[138] Mcaffrey AP, Meuse L, Pham TT,et al. RNA inteference in adult mice.Nature, 2002; 418:38-39.
[139] Brummelkamp TR, Bernards R, Agami R. Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell, 2002; 2:243-247.
[140] Brummelkamp TR, Bernards R, and Agami R. A system for stable expression of short interfering RNAs in mammalian cells. Science, 2002;296:550-553.
[141] Mittal V. Improving the efficiency of RNA interference in mammals. Nat Rev Genet, 2004; 5:355-365.
[142] Van de Wetering M, Oving I, Muncan V, et al. Specific inhibition of gene expression using a stably integrated, inducible small-interfering-RNA vector. EMBO Rep, 2003; 4:609-615.
[143] Zhang L, Fogg DK, Waisman DM. RNA interference-mediated silencing of the S100A10 gene attenuates plasmin generation and invasiveness of Colo 222 colorectal cancer cells. J Biol Chem, 2004;279:2053-2062.
[144] Li hua,Weiquan Liu,Taiyi Wang,et al.A Comparative Study on the method of Lipofection and Calcium Phosphate Transfection.Chi Jour of Lab Ani Sci,2000;10:65-68.
[145] Xia H,et al.SiRNAs mediated gene silencing i n vi t ro and in vivo.Nat Biotech,2002;20:1006.
[146] Hasuwa H,et al.Small interfering RNA and gene silencing in transgenic mice and rats.FEBS Lett,2002,532:227.
[147] Ubinson DA,et al.A lentivirus2based system to functionally silence genes in primary mammalian cells,stem cells and t ransgenic mice by RNA interference.Nat Genet,2003;33:401.
[148] McCaffrey A P,et al.RNA interference in adult mice.Nature,2002;418:38.
[149] Shi Y.Mammalian RNAi for the masses.Trends Genet,2003;19:9-12.
[150] Wang Lu,Hong Li,Yulin Li,et al.Expression of MMp-9 and MMP-9 mRNA in gastric carcinoma and its correlation with angiogenesis. Nat Med Jou of Chi,2003;83:782-786.
[1] 刘飞德,陈凛,李丽,等.人胃癌鸡胚移植模型的建立及其肿瘤生物学研究[J].消化外科,2004,3(2):114-117.
[2] 郑青.蒙古沙土鼠幽门螺杆菌感染性胃炎和胃癌模型[J].国外医学消化系疾病分册,2000,20(3):131.
[3] 姜福义,石艳华,杨秀峰,等.医学实验中实验动物的选择[J].承德医学院学报,1999,16(4):400-401.
[4] 徐晶莹,李益民,李玉兰,等.MNNG 诱导大鼠胃癌模型的建立[J].哈尔滨工业大学学报,2003,(37):104-106.
[5] Lu YH,Li YHn,Dong GG,et al.Multifactor constructed stomach cancer models on rat[J].J Har Med Univ,2005,39(3):226-227.
[6] Watenable T,Tada M,Nagai H,et al.Helicobacter pylori infection induces gastric cancer in mongolian gerbils[J].Gastroenterology,1998,115(3):642-648.
[7] Drazek ES , Dubois A , Holmes RK.Characterization and pre-sumptive identification of Helicobacter pylori isolates from rhe-sus monkeys[J] . Clin Microbiol,1994,32(7):1799-1804.
[8] Lv Xiao-ying,SU Mian-cheng,LI You,et al.Effect of Acanthopansx giraldii Hanns Var. Hispidus Hoo polysaccharides on the human gastric cancer cell line SGC-7901 and its possible mechanism [J].Chin Med J,2002,115(5):716-721.
[9] 刘家仁,陈丙卿,杨艳梅,等.β-紫罗兰酮抑制 SCC-7901 细胞潜在的转移作用[J].卫生研究,2005,34(4):435-438.
[10] 时洪凯,王毅,陈燕,等.小鼠结直肠自发肿瘤模型的建立[J].现代生物医学进展,2007, 7(5):714-717.
[11] 耿敬姝,薛英威,刘秀华,等.人消化道癌裸鼠移植瘤株及体外细胞系的建立[J].实用肿瘤学杂志,2000,14 (3):162- 164.
[12] 刘平,赵志泉,张小勇,等.人胃癌细胞悬液裸小鼠原位移植模型建立及其部分生物学特性[J].南京医科大学学报,2000,20(6):415- 416.
[13] 陶凯雄,田源,陈道达,等.鼠种植性胃肿瘤模型的制作方法及抗癌药物的治疗作用[J].同济医科大学学报,2000,29(3):229 -231.
[14] 陈亚琳,魏品康,许玲,等.采用 OB 胶粘贴法建立人胃癌裸鼠原位种植转移模型[J].癌症,2005, 24(2):246-249.
[15] He Y,Li YJ.Study on method of establishment of laboratory animal model of gastric cancer[J].Acta Lab Ani Sci Sin,2006,14(4):251-254.
[16] Duncan MD,Tihan T,Donovan DM,et al.Esophagogastric adenocarcinoma in an E1A /E1B transgenic model involves p53 disruption[J].Gastrointest Surg,2000,4(3):290 -297.
[17] Maruyama C,Tomisawa M,Wakana S,et al.Overexpression of human H-ras transgene is responsible for tumors induced by chemical carcinogens in mice[J].Oncol Rep,2001,8 (2):233 -237.
[18] Touati E,Hofnung M,Thiberge JM,et al.Short-term infection with Helicobacter pylori and 1 week exposure to metronida- zole does not enhance gastric mutation frequency in trans-genic mice[J].Antimicrobiol Chemother,2000,46(6):987-992.
[19] Kanai M,Konda Y,Nakajima T,et al.TGF-alpha inhibits apoptosis of murine gastricpit cells through a NF-kappaB-dependent pathway[J].Gastroenterology,2001,121(1):56-67.
[20] 李育超,周克元.非环式维甲酸类化学预防肝癌的研究进展[J].国际检验医学杂志,2006, 27(5):413-415.
[21]司建华,谭家驹,陈凤兰,等.亚硝化农肥水诱发鸡咽食管癌、胃癌和肝癌的病理形态学研究[J].青海医药杂志,2006,36(5):1-4.
[22] 朱人敏, 江绍基,萧树东,等.ENNG 诱发犬胃癌过程细胞动力学的研究[J].医学研究生学报,1992,5(4):357-360.
[23] Kalinovskii VP,Volfson NI,Petrov AS,et al.Changes in the peps in activity of the gastric mucosa of rats in the early stages of carcino genesis[J].Vopr Onkol,1985,31(12):76-80.
[2] Matvey E,Lukashev,Zena Werb,et al.ECM signalling: orchestrating cell behaviour and misbehaviour.Trends in Cell Biology, 1998;8:437-441.
[3] The role of gelatinases in colorectal cancer progression and metastasis. Biochimica et Biophysica Acta (BBA) - Reviews on Cancer.2004;1705:69-89.
[4] Kenny HA,Kaur S,Coussens LM,et,al.The initial steps of ovarian cancer cell metastasis are mediated by MMP-2 cleavage of vitronectin and fibronectin.J Clin Invest.2008;118:1367-1379.
[5] Visse R,Nagase H.Matrix Metalloproteinases and Tissue Inhibitors of Metalloroteinases:Structure,Function,and Biochemistry.Circulation Research,2003;92:827-839.
[6] Brew K,Dinakarpandian D,Nagase H.Tissue inhibitors of metalloproteinases:evolution,structure and function.Biochim Biophys Acta,2000;1477:267-283.
[7] Yoon SO,Park SJ,Yun CH,et al.Roles of Matrix Metalloproteinases in Tumor Metastasis and Angiogenesis.Journal of Biochemistry and Molecular Biology,2003;36:128-137.
[8] Bogenrieder T,Herlyn M.Axis of evil:molecular mechanisms of cancer metastasis,Oneogene,2003;22:6524-6536.
[9] Woessner JF Jr.Matrix metalloproteinases and their inhibitors in connective tissue remodeling.FASEB J,1991;5:2145-2154.
[10] Connor CMO,Fitz Gerald MX.Matrix metalloproteinases and lung disease.Thorax,1994;49:602.
[11] Nagase H,Woessner JF Jr.Matrix metalloproteinases.J Biol Chem.1999;2742:1491-1494.
[12] Krejci P,Masri B,Fontaine V,et al.Interaction of fibroblast growth factor and C-natriuretic peptide signaling in regulation of chondrocyte proliferation and extracellular matrix homeostasis.J Cell Sci. 2005;118:5089-5100.
[13] Birkedal Hansen H,Moore WG I,Bodden,et a1.Matrixmetalloproteinases.Crit Rev Oral Biol Med,1993;4:97.
[14] LI Jing,ZHAO Tianfu,DUAN Enkui.Matrix metalloproteinases (MMPs) and trophoblast invasion.science report,2005;12:1169-1173.
[15] Rhee JS,Coussens LM.RECKing MMP function:implications for cancer development.Trends Cell Biol.2002;12:209-211.
[16] Huang H, Wang M. Advances in the Molecular Mechanisms of Tumor Infiltration and Metastasis.Letters in biotechnology,2006 ;7: 54-56.
[17] Liotta LA, Steeg PS, Stetler-Stevenson WG.Cancer metastasis and angiogenesis:an imbalance of positive and negative regulation.Cell,1991;64:327-336.
[18] Crawford, H.c.Martrisian,et al.L.M Tumor stromal expression of matrixmetall oproteinases their role in tumor progression.Invasion metastasis,l994;14:234-245.
[19] Stetler-stervenson,W.G Aznavoortan,S Liotta,LA,et al.Tumor cell interaction with the extra cellular matrix during invasinon and metastasis.Annu Rev Cell Biol, 1993;9:541-573.
[20] Liotta LA,Tryggvason K,Garbisa S,et al.Metastatic potential correlates with enzymatic degradation of basement membrane collagen.Nature,1980;284:67-68.
[21] DaviesB,MilesDW,Happerfield LC,et al.Levels of type IV collagenases in benign and malignant breast disease.Br J Cancer Res,1993;67:1126-1131.
[22] Davies B,Waxman J,Wasan H,et al.Levels of Matrixmetalloproteinases in bladder cancer correlate with tumor grade invasion.Cancer Res,1993;53:5365-5369.
[23] Chambers AF,Matrisian L,et al.Changing views of the role of matrMetall opro- teinases in metastasis.Natl Cancer Inst.1997;89:1260-1270.
[24] Kahari VM,Saraarial ho-KereU,et al,matrixmetalloproteinases and the Inhibitors in tumor growth invasion.Ann Med,1999;31:34-45.
[25] Ray JM,Stetler Stevenson WG,et al.Gelatinase A activity directly modulate Mela- noma cell adhesion spreading.EMBO,1995;14:908-917.
[26] Kleiner DE,Stetler Stevenson WG.Matrixmetalloproteinases and metastasis. Can Che Pha,1999;43:342-351.
[27] Sauter W,Rosenberger A,Beckmann L,et al.Matrix Metalloproteinase 1(MMP1) Is Associated with Early-Onset Lung Cancer.Cancer Epidemiol Biomarkers Prev.2008;17:1127-1135.
[28] Alfano RW,Leppla SH,Liu S,et al.Cytotoxicity of the matrix metalloproteinase-activated anthrax lethal toxin is dependent on gelatinase expression and B-RAF status in human melanoma cells.Mol Cancer Ther.2008;7:1218-1226.
[29] De Cicco C,Ravasi L,Zorzino L,et al.Circulating levels of VCAM and MMP-2 may help identify patients with more aggressive prostate cancer.Curr Cancer Drug Targets.2008;8:199-206.
[30] Bendrik C,Robertson J,Gauldie J,et al.Gene transfer of matrix metalloproteinase-9 induces tumor regression of breast cancer in vivo.Cancer Res.2008;68:3405-1342.
[31] Lakka SS,Gondi CS,Yanamandra N,et al.Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis.Oncogene,2004;23:4681-4689.
[32] Ito H,Duxbury M,Benoit E,et al.Fibronectin-induced COX-2 mediates MMP-2 expression and invasiveness of rhabdomyosarcoma.Biochem Biophys Res Commun.2004;318:594-600
[33] Marin-Casta?o ME, Csaky KG, Cousins SW.Nonlethal oxidant injury to human retinal pigment epithelium cells causes cell membrane blebbing but decreased MMP-2 activity.Invest Ophthalmol Vis Sci. 2005 ;46:3331-3340.
[34] Karagiannis ED, Popel AS. Distinct modes of collagen type I proteolysis bymatrix metalloproteinase (MMP) 2 and membrane type I MMP during the migration of a tip endothelial cell: insights from a computational model.J Theor Biol. 2006;238:124-145.
[35] Bai YL, Huang HC, Li JZ, Zhao YY, Wang HY.High glucose regulates the production of MMP-9 in podocyte through ERK1/2 signal pathway.Zhonghua Yi Xue Za Zhi,2005;85:1451-1455.
[36] Azkur AK, Kim B, Suvas S, Lee Y, Kumaraguru U, Rouse BT. Blocking mouse MMP-9 production in tumor cells and mouse cornea by short hairpin (sh) RNA encoding plasmids.Oligonucleotides. 2005;15:72-84
[37] Giraudo E, Inoue M, Hanahan D.An amino-bisphosphonate targets MMP-9-expressing macrophages and angiogenesis to impair cervical carcinogenesis.J Clin Invest.2004;114:623-633.
[38] Topp SA, Upadhya GA, Strasberg SM. Cold preservation of isolated sinusoidal endothelial cells in MMP 9 knockout mice: effect on morphology and platelet adhesion.Liver Transpl. 2004;10:1041-1048.
[39] Liotta LA,Tryggvason K,Garbisa S,et a1.Metastatic potential correlates with enzymatic degradation of basement membrane collagen.Nature,1980;284:67
[40] Albini A,Melchiori A,Santi L,et a1.Tumor cell invasion inhibited by TIMP-2. Natl Cancer Inst,1991;83:775.
[41] Kawamata H,Kameyama S,Kawai K,et a1.Marked acceleration of the metastatic phenotype of a rat bladder carcinoma cell line by the expression of human gelatinase A.Int J Cancer,1995;63:568.
[42] Kawamata H,Kawai K,Kameyama S,et al. Over-expression of tissue inhibitor of matrix metalloproteinases (TIMP1 and TIMP2) suppresses extravasation of pulmonary metastasis of a rat bladder carcinoma.Int J Cancer,1995;63:680.
[43] Ray JM,Stetler Stevenson WG.The role of matrix matalloproteases and their inhibitors in tumour invasion,metastasis and angiogenesis.Eur Respir J,1994;7:2062.
[44] Yoshikawa T,Tsuburaya A,Kobayashi O,et al.Intratumoral concentrations oftissue inhibitor of matrix metalloproteinase Iin patients with gastric carcinoma a new biomartker for invasion and its impact on survival.Cancer,2001;91:1739.
[45] Garzetti G,Andrea C,Guendalina L,et a1.Expression of vascular endothelial growth factor related to 72-Kiloalton metalloproteinase immunostaining patients with seriousovarian tumor.Cancer,1999;85:2219-2225.
[46] Schwartz GK,Warg H,Lampen N,et a1.Defining the invasive phenol type of proximal gastric cancer cells.Cancer,1994;73:222-227.
[47] Li nan,Caipu Xu,Weiwen Lu,et al.Effect of tissue inhibitor of matrix metalloproteinase-2 spontanous invasion of gastric carcinoma cell lines in vitro. China Oncology,2000;10:163-165.
[48] Li nan,Caipu Xu,Weiwen Lu,et al.Effect of tissue inhibitor of matrix metalloproteinase-2 spontanous invasion of gastric carcinoma cell lines in vitro and in vivo.China Oncology .Chinese Journal of Pathology,1998;27:362-365.
[49] Inoue T,Yashiro M,Nishimra S.Matrix metalloproteinases1 expression is a prognostic factor for Patients with advanced gastric cancer.In J Mol-Med, 1999;4:73-77.
[50] Li L,Zhang sheng,Lin hua,et al.Relationship of Expression Unbalance of Matrix Metalloproteinase and Tissue Inhibitor of Metalloproteinase to Invasiveness and Metastasis in Gastric Carcinomas. Chi Jou of Can,2002;21:305-310.
[51] Yoshikawa T,Tsuburaya A,KobayashiO,et al.Prognostic value of tissue inhibitor of matrix metalloproteinase-I in plasma of patients with gastric cancer.Cancer Lett,2000;151:81-86.
[52] Zhang S,Li L,Lin J, et al.Imbalance between expression of matrix metalloproteinase-9 and tissue inhibitor of metaloproteinase-1 in invasiveness and metastasis of human gastric carcinoma.Wor J Gas,2003;9:899-904.
[53] Yutaka Takahashi,Yasuhiko Kitadai,Lee M.et al.Multiparametric in situmRNA Hybridization Analysis of Gastric Biopsies Predicts Lymph Node Metastasis in Patients with Gastric Carcinoma.Cancer Res,2002;93:1258-1265.
[54] Xiaoping Miao, Chunyuan Yu,Wen Tan,et al.A Functional Polymorphism in the Matrix Metalloproteinase-2 Gene Promoter Is Associated with Risk of Development but not Metastasis of Gastric Cardia Adenocarcinoma. Cancer Research,2003;63:3987-3990.
[55] Matsumura S,Oue N,Nakayama H,et al.A single nucleotide polymorphism in the MMP-9 promoter affects tumor progression and invasive phenotype of gastric cancer.Can Res Clin Onco,2005;131:19-25.
[56] Koyama S.Enhanced cell surface expression of inhibitors,and tumor-induced host response matrix metalloproteinases and their in progression of human gastric carcinoma.Dig Dis Sci,2004;49:1621-1630.
[57] Murray GI,Duncan ME,Arbuckle E,Melvin WT,Fothergill JE.Matrix metalloproteinases and their inhibitors in gastric cancer.Gut,1998;43:791-797.
[58] Tsuchiya Y,Sato H,Endo Y,et al.Tissue inhibitor of metalloproteinase 1 is a negative regulator of the metastatic ability of a human gastric cancer cell line, KKLS, in the chick embryo.Cancer Res,1993;53:1397-402.
[59] Nomura H,Fujimoto N,Seiki M,et al.Enhanced production of matrix metalloproteinases and activation of matrix metalloproteinase 2 (gelatinase A) in human gastric carcinomas.Int J Cancer,1996;69:9-16.
[60] Lisa M.C,Barbara Fingleton,and Lynn M.Matrisian Matrix Metalloproteinase Inhibitors and Cancer:Trialsand Tribulations.Science,Mar,2002;295:2387-2392.
[61] Sang-Oh Yoon,Soo-Jin Park,Chang-Hyun Yun,eta1.Roles of in Tumor Matastasis and Angiogenesis.Journal of Biochemistry and Molecular Biology,2003;36:128-137.
[62] Price A,SW Q,Morris D,et al.Marked inhibition of tumor growth in amalignant glioma tumor model by a novel synthetic matrixmetalloproteinase inhibitorAG3340.Clin Cancer Res,1999;5:845-854.
[63] Knox JD,Bretton L,Lynch et al.Synthetic matrix metalloproteinase inhibitor,BB-94,inhibits the invasion of neoplastic human prostate cells in a mouse model.Prostate,1998;35:248-254.
[64] Anderson IC,Shipp MA,Docherty AJ,et al.Combination therapy including a gelatinase inhibitor and cytotoxic agent reduces local invasion and metastasis of murine Lewis lung carcinoma.Cancer Res,1996;56:715-718.
[65] Nicoletti V,Lucchini M,D'Incalci R,et a1.Comparison of paclitaxel and docetaxel activity on human ovarian carcinoma xenografts.European Journal of Cancer, 1994;30:691-696.
[66] Godehard Friedel,Ugo Pastorino,Robert J,et al.Results of lung metastasectomy from breast cancer: prognostic criteria on the basis of 467 cases of the international registry of lung metastases. European Journal of Cardio-Thoracic Surgery,2002;22:335-344.
[67] Bodurtha A,Eisenhauer E,Steward W,et al.Phase I-II study of marimastat(BB-2516) in patients with metastatic melanoma.Proc Am Soc Clin Oncol,1997;16:493.
[68] Parsons SL,Watson SA,Griffin NR,et a1.An open phase I/II study of the oral matrix metalloproteinase inhibitor marimastat in patients with inoperable gastric cancer.Ann Oncol,1996;47:47.
[69] Parano JA,Bernardo P,Stephenson P,et al.Randomized phase III trial of marimastat versus placebo in patients with metastatic breast cancer who have responding or stable disease after first-line chemotherapy.Clin Oncol,2004;22:4683-4690.
[70] Nemunaitis J,Poole C,Primrose J,et al.Combined analysis of studies of the effects of the matrix metalloproteinase inhibitor marimastat on serum tumor markers in advanced cancer:Selection of abiologically active and tolerable dose for longer-term studies.ClinCancerRes,1998;4:1101-1109.
[71] Rosemurgy A,Buckels J,Charnley R.Arandomized study comparing marimastat to gemcitabine as first line therapy in patients withnon-resectable pancreatic cancer.Proc Am Soc Clin Oncol,1999;18:261.
[72] Gradishar W,Sparano J,Cobleigh M,et al,A phase I study of marimastat in combination with doxorubicin and cyclophosphamide in patients with metastatic breast cancer.Proc Am Soc Clin Onco1,1998;17:144.
[73] Wildingq Small E,Rippleq et al,Phase I study of AG3340,a matrix metalloproteinase inhibitor incombination with mitoxantronel prednisonein patients having advanced prostate cancer.Ann Oncol,1998;9:74.
[74] Ikeda T,Murakami K,Hayakawa Y,et al.Anti-invasive activity of synthetic serine protease inhibitors and its combined effect with a matrix metalloproteinase inhibitor.Anti cancer Res,1998;18:4259-4265.
[75] Fire A,Xu S,Montgomery MK,et al.Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans.Nature,1998;391:806-811.
[76] Cogoni C,Macino G.Posttranscriptional gene silencing in Neurospora by a RecQ DNA helicase.Science,1999;286:2342-2344.
[77] Hannon GJ.RNA interference. Nature ,2002;418:244-251.
[78] Mario Stevenson.Dissecting HIV-1 through RNA interference.Nat Rev Immunol,2003;3:851-858.
[79] Elbashir SM,et al. Duplexes of 21 nucleotide RNAs mediate RNA interference in culture mammalian cells.Nature,2001;411:494-498.
[80] Elbashir SM,et al. RNA interference is mediated by 21-and 22-nucleotide RNAs .Genes Dev,2001;15:188-200.
[81] Verma NK,et al.RNA-mediated gene silencing:mecha-nisms and its therapeutic applications.J Clin Pharm T-her.2004,29:395-404.
[82] Angela Reynolds,et al.Rational siRNA design for RNA in-terference. Nature biotechnology,2004;22:326-330.
[83] Phillip D,et al.Ancient pathways programmed by small RNA.Science,2002;296:1265-1269.
[84] Ronald HAP. RNA silencing:the genome,s immune system.Science,2002;296:1263-1265.
[85] Michael B,et al. Genome-wide RNAi analysis of growth and viability in drosophila cells. Science,2004;303:832-835
[86] Verma NK,et al. RNA-mediated gene silencing:mechanisms and its therapeutic applications.J Clin Pharm T-her.2004;29:395 - 404.
[87] McCaffrey AP,et al. RNA interference in adult mice.Nature,2002;418:38-39.
[88] Kronke J,et al. Alternative approaches for efficient in-hibition of hepatitis C virus RNA replication by small in-terfering RNAs. J Virol,2004;78:3436-3446.
[89] Takigawa Y,et al.Suppression of hepatitis C virus replicon by RNA interference directed against the NS3 and NS5B regions of the viral genome.Microbiol Immunol,2004;48:591-598.
[90] George JA, Gene therapy progress and prospects:adenoviral vectors,Gene Therapy,2003;6:12-16.
[91] Liotta LA,Kohn EC,Cancer's deadly signature.Nat Genet,2003;33:49-54
[92] Li yingping,Zhang ying,Shao guoqing,A New Technology to Delete Gene-RNAi,Bio Bull,2006;3:27-30.
[93] Marjori M,et al.RNA:guiding gene silencing. Science,2001;293:1080-1083.
[94] Verma NK,et al.RNA-mediated gene silencing:mechanisms and its therapeutic applications.J Clin Pharm Ther.2004;29:395-404.
[95] Jun Yuan A, Charyl M,Dutton B,Sean P,et al.RNAi mediated MMP-1 silencing inhibits human chondrosarcoma invasion.Jou of Orth Res,2005;23:1467-1474.
[96] Brummelkamp TR,Bernards R,Agami R.Stable suppression of tumor igenicity by virus-mediated RNA interference.Cancer Cell,2002;2:243-247.
[97] Fleming JB,Shen GL,Holloway SE,et al.Molecular consequences of silencing mutant K-ras in pancreatic cancer cells:justification for K-ras-directed therapy.Mol Cancer Res,2005;3:413-423.
[98] Wang YH,Liu S,Zhang G,et al.Knockdown of c-Myc expression by RNAi inhibits MCF-7 breast tumor cells growth in vitro and in vivo.Breast CancerRes,2005;7:220-228.
[99] Takada Y,Sethi G,Sung B,et al.Flavopiridol suppresses tumor necrosis factor-induced activation of activator protein-1,c-Jun N-terminal kinase,p38 mitogen-activated protein kinase (MAPK),p44/p42 MAPK,and Akt,inhibits expression of antiapoptotic gene products,and enhances apoptosis through cytochrome c release and caspase activation in human myeloid cells.Mol Pharmacol.2008;73:1549-1557.
[100] Won KA,Reed SI.Activation of cyclin E/CDK2 is coupled to site-specific autophosphorylation and ubiquitin-dependent degradation of cyclin E.EMBO J.1996;15:4182-4193.
[101] Terano T,Tanaka T,Tamura Y,et al.Eicosapentaenoic acid and docosahexaenoic acid inhibit vascular smooth muscle cell proliferation by inhibiting phosphorylation of Cdk2-cyclinE complex.Biochem Biophys Res Commun.1999;254:502-506.
[102] Li K,Lin S,Brunicardi FC,et al.Use of RNA interference to target cyclin E over expressing hepatocellular carcinoma.Cancer Res,2003,63:3593-3597.
[103] Martin-Lluesma S, Stucke VM,Nigg EA.Role of Hec1 in spin-dle checkpoint signaling and kinetochore recruitment of Mad1/Mad2.Science,2002;297:2267-2270.
[104] Chen Z,Varney ML,Backora MW,et al.Down-regulation of vascular endothelial cell growth factor-C expression using small interfering RNA vectors in mammary tumors inhibits tumor lymphangiogenesis and spontaneous metastasis and enhances survival.Cancer Res,2005;65:9004-9011.
[105] Shen J,Samul R,Silva RL,et al.Suppression of ocular neovascularization with siRNA targeting VEGF receptor 1.Gene Ther,2006;13:225-234.
[106] Amantana A,London CA, Iversen PL,et al.X-linked inhibitor of apoptosis protein inhibition induces apoptosis and enhances chemotherapy sensitivity in human prostate cancer cells. Mol Cancer Ther,2004;3:699-707.
[107] Stierle V,Laigle A,Jolles B.Modulation of MDR1 gene expression inmultidrug resistant MCF7 cells by low concentrations of small interfering RNAs.Biochem Pharmacol,2005;70:1424-1430.
[108] .Pichler A,Zelcer N,Prior JL.In vivo RNA interference-mediated ablation of MDR1 P-glycoprotein.Clin Cancer Res,2005;11:4487-4494.
[109] Kami k,Doi R,Koizumi M,et al.Downregulation of surviving by siRNA diminishes radioresistance of pancreatic cancer cells.Surgery,2005;138:299-305.
[110] Tuschl T,Zamore PD,Lehmann R,et al.Targeted mRNA degradation by double-stranded RNA in vitro.Genes Dev,1999;13:3191-3197.
[111] Bernstein E,Caudy AA,Hammond SM,Hannon GJ.Role for a bidentate ribonuclease in the initiation step of RNA interference.Nature,2001;409:363-366.
[112] Zamore PD,Tuschl T,Sharp PA,Bartel DP.RNAi:double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals.Cell,2000;101:25-33.
[113] Paddison PJ,Caudy AA,Bemstein E,et a1.Short hairpin RNAszz (shRNAs)induce sequence-specific silencing in mammalian cells.Genes 2002;16:948-958.
[114] Miyagishi M,Taira K.U6 promoter-driven siRNAs with four uridine 3’overhangs efficiently suppress,targeted gene expression in mammalian cells.Nat Biotech,2002;19:497-500.
[115] Brummelkamp TR,Bernads R,Agami R.A system for stable expression of short interfering RNAs in mammalian cells.Science,2002;296:550-553
[116] Sachidanandam R.RNAi:design and analysis.Curr Protoc Bioinformatics.2004;12:Unit 12.3
[117] Kuittinen O,Soini Y,Turpeenniemi-Hujanen T.Diverse role of MMP-2 and MMP-9 in the clinicopathological behavior of Hodgkin's lymphoma.Eur J Haematol.2002;69:205-212.
[118] von Lüttichau I,Djafarzadeh R,Henger A,et al. Identification of a signal transduction pathway that regulates MMP-9 mRNA expression inglomerular injury.Biol Chem.2002;383:1271-1275.
[119] Lahat N,Shapiro S,Froom P,et al.Inorganic lead enhances cytokine-induced elevation of matrix metalloproteinase MMP-9 expression in glial cells.J Neuroimmunol.2002;132:123-128.
[120] Fang J,Shing Y,Wiederschain D,et al.Matrix metalloproteinase-2 is required for the angiogenic phenotype in a tumormodel.Proc Natl Acad Sci USA,2000;97:3884.
[121] Curran S,Murray GI.Matrix metalloproteinases in tumour invasion and metastasis.J Pathol,1999;189:300-308.
[122] Coussens LM,Fingleton B,Matrisian LM.Matrix metalloproteinases,inhibitors and cancer:trials and tribulations.Science,2002;295:2387-2391.
[123] Itoh T,Tanioka M,Yoshida H,et al.Reduced angiogenesis and tumor progression in gelatinase Adeficient mice.CancerRes,1998;58:1048-1051.
[124] NelsonAR,Fingleton B,Rothenberg ML,MatrisianLM.Matrix metal oproteinases:biologicactivity and clinicali mplications.J Clio Oncol, 2000;18:1135-1149.
[125] Schmalfeldt B,Prechtel D,Harting K,et al.Increased expression of matrixmetalloproteinases(MMP)-2,MMP-9,andtheurokinase-type plasminogen activator is associated with progression from benign to advancedovarian cancer.Clin Cancer Res,2001;7:2396-404.
[126] Yanagisawa N, Geironson L, Waleed AS, et al. Expression of matrix metalloprotease-2,-7,and -9 on human colon, liver and bile duct cell lines by enteric and gastric Helicobacter species.FEMS.Immunol Med Microbiol,2005;44:197-204.
[127] Sier CF,Kabben FJ,Ganesh S,et al.Tissue levels of matrix metalloproteinases MMP-2 and MMP-9 are related to the overall survival of patients with gastric carcinoma.Br J Cancer,1996;74:413-417.
[128] Wu ZY, Li JH, Zhan WH, et al. Lymph node micrometastasis and its correlation with MMP-2 expression in gastric carcinoma.World JGastroenterol, 2006;12:2941-2944.
[129] Sham S K,Mohammad TM.Regulation of angiogenesis and invasion by human pituitary tumor transforming gene(PTTG) through increased expression and secretion of matrix metalloproteinase-2 (MMP-2).Molecular Cancer,2006;5:61.
[130] Nomura H,Sato H,Seiki M,et al.Expression of membrane type matrix metalloproteinase in human gastric carcinomas.Cancer Res,1995;55:3263-3266.
[131] Nomura H,Fujimoto N,Seiki M,et al.Enhanced production of matrix metalloproteinases and activation of matrix metalloproteinase 2 (gelatinaseA)in human gastric carcinomas.Int J Cancer,1996;69:9-16.
[132] Kubben FJ,Sier CF,van Duijn W,et al.Matrix metalloproteinase-2 is a consistent prognostic factor in gastric cancer.Br J Cancer,2006;10,94:1035-1040.
[133] Chang C,Werb Z. The many faces of metalloproteases: cell growth, invasion, angiogenesis and metastasis.Trends Cell Biol,2001;11:S37-S43.
[134] Tuschl T. Expanding small RNA interference. Nat Biotechnol, 2002; 20:446-448.
[135] McManus MT, Sharp PA. Gene silencing in mammals by small interfering RNAs. Nat Rev Genet, 2002;3:737-747.
[136] Wang J, Tekle E, Oubrahim H, et al. Stable and controllable RNA inteference:investigating the physiological function of glutathionylated actin. Proc Natl Acad Sci USA, 2003;100:5103-5106.
[137] Zhou bo,Yang wei,Jianguo Ji,et al.Differential Protein Expression Induced by Transient Transfection of Metallothionein-3 Gene in SH-SY5Y Neuroblastoma Cell LineDifferential Protein Expression Induced by Transient Transfection of Metallothionein-3 Gene in SH-SY5Y Neuroblastoma Cell Line. ACTA BIOCHIMICA ET BIOPHYSICA SINICA. 2003;6:522-528.
[138] Mcaffrey AP, Meuse L, Pham TT,et al. RNA inteference in adult mice.Nature, 2002; 418:38-39.
[139] Brummelkamp TR, Bernards R, Agami R. Stable suppression of tumorigenicity by virus-mediated RNA interference. Cancer Cell, 2002; 2:243-247.
[140] Brummelkamp TR, Bernards R, and Agami R. A system for stable expression of short interfering RNAs in mammalian cells. Science, 2002;296:550-553.
[141] Mittal V. Improving the efficiency of RNA interference in mammals. Nat Rev Genet, 2004; 5:355-365.
[142] Van de Wetering M, Oving I, Muncan V, et al. Specific inhibition of gene expression using a stably integrated, inducible small-interfering-RNA vector. EMBO Rep, 2003; 4:609-615.
[143] Zhang L, Fogg DK, Waisman DM. RNA interference-mediated silencing of the S100A10 gene attenuates plasmin generation and invasiveness of Colo 222 colorectal cancer cells. J Biol Chem, 2004;279:2053-2062.
[144] Li hua,Weiquan Liu,Taiyi Wang,et al.A Comparative Study on the method of Lipofection and Calcium Phosphate Transfection.Chi Jour of Lab Ani Sci,2000;10:65-68.
[145] Xia H,et al.SiRNAs mediated gene silencing i n vi t ro and in vivo.Nat Biotech,2002;20:1006.
[146] Hasuwa H,et al.Small interfering RNA and gene silencing in transgenic mice and rats.FEBS Lett,2002,532:227.
[147] Ubinson DA,et al.A lentivirus2based system to functionally silence genes in primary mammalian cells,stem cells and t ransgenic mice by RNA interference.Nat Genet,2003;33:401.
[148] McCaffrey A P,et al.RNA interference in adult mice.Nature,2002;418:38.
[149] Shi Y.Mammalian RNAi for the masses.Trends Genet,2003;19:9-12.
[150] Wang Lu,Hong Li,Yulin Li,et al.Expression of MMp-9 and MMP-9 mRNA in gastric carcinoma and its correlation with angiogenesis. Nat Med Jou of Chi,2003;83:782-786.
[1] 刘飞德,陈凛,李丽,等.人胃癌鸡胚移植模型的建立及其肿瘤生物学研究[J].消化外科,2004,3(2):114-117.
[2] 郑青.蒙古沙土鼠幽门螺杆菌感染性胃炎和胃癌模型[J].国外医学消化系疾病分册,2000,20(3):131.
[3] 姜福义,石艳华,杨秀峰,等.医学实验中实验动物的选择[J].承德医学院学报,1999,16(4):400-401.
[4] 徐晶莹,李益民,李玉兰,等.MNNG 诱导大鼠胃癌模型的建立[J].哈尔滨工业大学学报,2003,(37):104-106.
[5] Lu YH,Li YHn,Dong GG,et al.Multifactor constructed stomach cancer models on rat[J].J Har Med Univ,2005,39(3):226-227.
[6] Watenable T,Tada M,Nagai H,et al.Helicobacter pylori infection induces gastric cancer in mongolian gerbils[J].Gastroenterology,1998,115(3):642-648.
[7] Drazek ES , Dubois A , Holmes RK.Characterization and pre-sumptive identification of Helicobacter pylori isolates from rhe-sus monkeys[J] . Clin Microbiol,1994,32(7):1799-1804.
[8] Lv Xiao-ying,SU Mian-cheng,LI You,et al.Effect of Acanthopansx giraldii Hanns Var. Hispidus Hoo polysaccharides on the human gastric cancer cell line SGC-7901 and its possible mechanism [J].Chin Med J,2002,115(5):716-721.
[9] 刘家仁,陈丙卿,杨艳梅,等.β-紫罗兰酮抑制 SCC-7901 细胞潜在的转移作用[J].卫生研究,2005,34(4):435-438.
[10] 时洪凯,王毅,陈燕,等.小鼠结直肠自发肿瘤模型的建立[J].现代生物医学进展,2007, 7(5):714-717.
[11] 耿敬姝,薛英威,刘秀华,等.人消化道癌裸鼠移植瘤株及体外细胞系的建立[J].实用肿瘤学杂志,2000,14 (3):162- 164.
[12] 刘平,赵志泉,张小勇,等.人胃癌细胞悬液裸小鼠原位移植模型建立及其部分生物学特性[J].南京医科大学学报,2000,20(6):415- 416.
[13] 陶凯雄,田源,陈道达,等.鼠种植性胃肿瘤模型的制作方法及抗癌药物的治疗作用[J].同济医科大学学报,2000,29(3):229 -231.
[14] 陈亚琳,魏品康,许玲,等.采用 OB 胶粘贴法建立人胃癌裸鼠原位种植转移模型[J].癌症,2005, 24(2):246-249.
[15] He Y,Li YJ.Study on method of establishment of laboratory animal model of gastric cancer[J].Acta Lab Ani Sci Sin,2006,14(4):251-254.
[16] Duncan MD,Tihan T,Donovan DM,et al.Esophagogastric adenocarcinoma in an E1A /E1B transgenic model involves p53 disruption[J].Gastrointest Surg,2000,4(3):290 -297.
[17] Maruyama C,Tomisawa M,Wakana S,et al.Overexpression of human H-ras transgene is responsible for tumors induced by chemical carcinogens in mice[J].Oncol Rep,2001,8 (2):233 -237.
[18] Touati E,Hofnung M,Thiberge JM,et al.Short-term infection with Helicobacter pylori and 1 week exposure to metronida- zole does not enhance gastric mutation frequency in trans-genic mice[J].Antimicrobiol Chemother,2000,46(6):987-992.
[19] Kanai M,Konda Y,Nakajima T,et al.TGF-alpha inhibits apoptosis of murine gastricpit cells through a NF-kappaB-dependent pathway[J].Gastroenterology,2001,121(1):56-67.
[20] 李育超,周克元.非环式维甲酸类化学预防肝癌的研究进展[J].国际检验医学杂志,2006, 27(5):413-415.
[21]司建华,谭家驹,陈凤兰,等.亚硝化农肥水诱发鸡咽食管癌、胃癌和肝癌的病理形态学研究[J].青海医药杂志,2006,36(5):1-4.
[22] 朱人敏, 江绍基,萧树东,等.ENNG 诱发犬胃癌过程细胞动力学的研究[J].医学研究生学报,1992,5(4):357-360.
[23] Kalinovskii VP,Volfson NI,Petrov AS,et al.Changes in the peps in activity of the gastric mucosa of rats in the early stages of carcino genesis[J].Vopr Onkol,1985,31(12):76-80.