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Frontiers in Biology 2007年第2期2卷 170-175页

作者： YAN Xunyou ZHAO Hongliang ZHANG Weiguang XUE Chong LIU Zhimin Insitute of Biotechnology Academy of Military Medical SciencesBeijing 100071China Biology Department Langfang Normal CollegeLangfang 065000China College of Food Science Southwest Agricultural UniversityChongqing 400716China

To obtain human tissue inhibitor of metalloproteinase-2(TIMP-2)cDNA and the secretory expression of TIMP-2 gene in Pichia pastoris,we designed and synthesized a 618 base pairs artificial gene coding for the TIMP-2 with a computer-aided design method using a standard chemical synthesis technique,which was composed of frequently used codons in the highly expressed Pichia pastoris *** the synthetic gene encoding TIMP-2 was checked by means of dideoxynucleotide *** verified gene of TIMP2 was cloned to the Escherichia coli-yeast shuttle vector of pPIC9 to construct a recombinant plasmid *** plasmid was transformed into GS115 cells of the methylotrophic yeast,Pichia pastoris by electroporation,and we got the expression cell through phenotype selection and induction with ***,purification,and bioactivity analysis of the expressed products were performed.

关键词： tissue inhibitor of metalloproteinase-2 Pichia pastoris expression bioactivity

Effects of benazepril on renal function and kidney expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in diabetic rats

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Chinese Medical Journal 2006年第10期119卷 814-821页

作者： SUN Shu-zhen WANG Yi LI Qian TIAN Yong-jie LIU Ming-hua YU Yong-hui Department of Pediatrics Shandong Provincial Hospital ShandongUniversity Jinan 250021 China

Background Excessive deposition of extraceUular matrix （ECM） in the kidney is the hallmark of diabetic nephropathy. Increased matrix synthesis has been well documented but the effects of diabetes on degradative pathways, particularly in the in vivo setting. The renal protective effect of these pathways on matrix accumulation has not been fully elucidated. The present study was understaken to investigate the activity of matrix metalloproteinase-2 （MMP-2）, the expression of MMP-2 and tissue inhibitor of metalloproteinase-2 （TIMP-2） in kidney tissues of diabetic rats, and to explore the degradative pathway of type Ⅳ collagen （Ⅳ-C） and the renal protective effects of ACE inhibition- benazepril.

关键词： angiotensin converting enzyme inhibitors diabetic nephropathy renal function matrix metalloproteinase-2 tissue inhibitor of metalloproteinase-2

Prognostic value of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in bladder carcinoma

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Chinese Medical Journal 2002年第5期115卷 743-745,155页

作者：许克新侯树坤杜志军北京大学人民医院泌尿外科北京100044

OBJECTIVES: To detect the level of matrix metalloproteinase-2 (MMP-2) mRNA and the tissue inhibitor of metalloproteinase-2 (TIMP-2) mRNA in bladder transitional cell carcinoma (BTCC), and to estimate the prognosis for bladder tumor based on the quality and quantity of MMP-2 and TIMP-2 mRNA. METHODS: Thirty-five samples of human BTCC and 15 normal fresh bladder tissues were studied by RT-PCR analysis followed by computer-assisted image analysis. RESULTS: The level of the MMP-2 mRNA in BTCC was significantly increased compared with that in normal bladder epithelium. The positive rates of MMP-2 and TIMP-2 mRNA were 71.4% and 65.7% in BTCC, and 66.7% and 60.0% in the normal bladder wall. The expression intensity of the MMP-2 mRNA by image analysis tended to increase with tumor grading and staging, which showed statistical significance. Similarly, the MMP-2 to TIMP-2 ratio also showed statistically significant difference between normal bladder tissue and bladder carcinoma (P

关键词： Adult Aged Aged, 80 and over Bladder Bladder Neoplasms Female Gelatinase A Gene Expression Regulation, Neoplastic Humans Male Middle Aged Neoplasm Staging Predictive Value of Tests Prognosis RNA, Messenger Research Support, Non-U.S. Gov't tissue inhibitor of metalloproteinase-2

The ratio of MMP-2 to TIMP-2 in hilar cholangiocarcinoma:a semi-quantitative study

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Hepatobiliary & Pancreatic Diseases International 2004年第4期3卷 599-602页

作者： Mei Xiao, Nin-Xing Zhou,Zhi-Qiang Huang,Ya-Li Lu, Ling-Hong Chen, Dian-Jun Wang and Wei-Lin Chang Beijing, China Department of Hepatobiliary Surgery, General Air- force of Hospital, Beijing 100036, China and General Hospital of People’ s Liberation Army, Beijing 100853 , China Department of Hepatobiliary Surgery of General Airforce of Hospital Beijing 100036 China. fengweiairforce@***

BACKGROUND: Hilar cholangiocarcinoma is associated with low resectability and poor survival. The aim of this study was to evaluate the roles of matrix metalloproteinases- 2 (MMP-2 ) and its tissue inhibitor of metalloproteinase-2 (TIMP-2) in tumor invasion or as a prognostic factor in patients with human hilar cholangiocarcinoma. METHODS: The expressions of MMP-2 and TIMP-2 were investigated in patients. Paraffinized tissue sections ob- tained from 50 patients with human hilar cholangiocarcino- ma were analysed. The expressions of MMP-2 and TIMP-2 were examined immunohistochemically. Image analysis with image-pro plus analysis software was used to semi- quantitatively determine the ratio of MMP-2 to TIMP-2. RESULTS: The expression levels of MMP-2 and TIMP-2 were strongly associated with tumor hepatic invasion in pa- tients with hilar cholangiocarcinoma. Significant diffe- rences in the ratio of MMP-2 to TIMP-2 between some pathologic factors were observed in patients with hilar cholangiocarcinoma. CONCLUSIONS: MMP-2 plays an essential role in tumor invasion and metastasis,while TIMP-2 is shown to strongly inhibit cancer invasion and metastasis. The ratio of MMP-2 to TIMP-2 may be a prognostic indicator for patients with hilar cholangiocarcinoma.

关键词： hilar cholangiocarcinoma matrix metalloproteinase-2 tissue inhibitor of metalloproteinase-2 immunohistochemical staining tumor invasion prognostic factor

Lichong decoction reduces Matrix metalloproteinases-2 expression but increases tissue inhibitors of Matrix metalloproteinases-2 ex-pression in a rat model of uterine leiomyoma

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Journal of Traditional Chinese Medicine 2016年第4期36卷 479-485页

作者： Wang Yasong Li Donghua Xu Xin Qian Ruiya Zhang Yalan Huang Yuhua Geng Jianguo Zou Xiaoli Han Hon-gjuan Zhang Wufang School of Traditional Chinese Medicine Capi-tal Medical Universityand School of General Practice andcontinuing EducationCapital Medical UniversityBeijing Xicheng District Desheng Community Health Service Cen-terBeijing 100120China School of Traditional Chinese Medicine Capital Medical UniversityBeijing Key Lab of TCM Collateral Disease Theory ResearchBeijing 100069China Department of Gynecology of Bei-jing Hospital of Traditional Chinese Medicine Beijing100010China The Fourth Ward of Beijing Ob-stetrics and Gynecology Hospjital Capital Medical Universi-tyBeijing Maternal and Child Health Care HospitalBeijing100026China

OBJECTIVE:To study the effect of Lichong decoction(LD) on expression of matrix metalloproteinase-2(MMP-2) and metalloproteinase-2(TIMP-2) in a rat model of uterine leiomyoma(UL).METHODS:UL was induced in rats using exogenous estrogen and *** was administered(p.o.) for 4 weeks,and mifepristone(RU-486)used as a *** observe the effect of LD on the uterine coefficient and uterine transverse diameter,a radioimmunoassay method was used to detect serum levels of sex *** microscopic analyses of pathologic changes in the tissues of UL rats were *** of the proteins of matrix metalloproteinases(MMPs) and tissue inhibitors of metalloproteinases(TIMPs) in uterine tissues was assessed by immunohistochemical staining and western ***:A UL model in rats was established *** reduced uterine weight,uterine coefficient,and uterine transverse diameter compared with untreated *** reduced levels of estradiol,progesterone,follicle-stimulating hormone,and luteinizing hormone in our UL *** improved the pathologic condition of uterine *** of MMP-2 protein decreased to varying extents in LD-treated groups,but TIMP-2 levels were *** appears to reduce MMP-2 expression and increase TIMP-2 expression in UL ***:These data suggest that the mechanism of action of LD on ULs may involve reduction of MMP-2 expression and increase in TIMP-2 expression in rats.

关键词： Leiomyoma Matrix metalloproteinase 2 tissue inhibitor of metalloproteinase-2 Lichong decoction

Effects of hypoxia,hyperoxia on the regulation of expression and activity of matrix metalloproteinase-2 in hepatic stellate cells

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World Journal of Gastroenterology 2001年第5期7卷 647-651页

作者： Ping-Sheng Chen~(1,2) Wei-Rong Zhai~1 Xiao-Mei Zhou~3 Jin-Sheng Zhang~1 Yue-E Zhang~1 Yu-Qin Ling~1 Ying-Hong Gu~1 1 Department of Pathology,School of Basic Medical Sciences,Fudan University,Shanghai 200032,China2 Ping-Sheng Chen now works in the Department of Pathology,School of Basic Medical Sciences the (former Nanjing Railway Medical College),Southeast University,Nanjing 210009,China3 Institute of Cancer Research,Shanghai 200032,China Department of Pathology School of Basic Medical Sciences Fudan University 138 Yixueyuan Road Shanghai 200032 China.

AIM: To study the effects of hypoxia, hyperoxia on the regulation of expression and activity of matrix metalloproteinase-2 (MMP-2) in hepatic stellate cells (HSC). METHODS: The expressions of MMP-2, tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and membrane type matrix metalloproteinase-1 (MT1-MMP) in cultured rat HSC were detected by immunocytochemistry (ICC) and in situ hybridization (ISH). The contents of MMP-2 and TIMP-2 in culture supernatant were detected with ELISA and the activity of MMP-2 in supernatant was revealed by zymography. RESULTS: In the situation of hypoxia for 12h, the expression of MMP-2 protein was enhanced (hypoxia group positive indexes: 5.7 +/- 2.0, n=10; control: 3.2 +/- 1.0, n = 7; P<0.05), while TIMP-2 protein was decreased in HSC (hypoxia group positive indexes: 2.5 +/- 0.7, n = 10; control: 3.6 +/- 1.0, n = 7; P < 0.05), and the activity (total A) of MMP-2 in supernatant declined obviously (hypoxia group: 7.334 +/- 1.922, n = 9; control: 17.277 +/- 7.424, n = 11; P < 0.01). Compared the varied duration of hypoxia, the changes of expressions including mRNA and protein level as well as activity of MMP-2 were most notable in 6h group. The highest value(A(hypoxia)-A(control)) of the protein and the most intense signal of mRNA were in the period of hypoxia for 6h, along with the lowest activity of MMP-2. In the situation of hyperoxia for 12h, the contents (A(450)) of MMP-2 and TIMP-2 in supernatant were both higher than those in the control, especially the TIMP-2 (hyperoxia group: 0.0499 +/- 0.0144, n = 16; control: 0.0219 +/- 0.0098, n = 14; P < 0.01), and so was the activity of MMP-2 (hyperoxia group: 5.252 +/- 0.771, n = 14; control: 4.304 +/- 1.083, n = 12; P < 0.05), and the expression of MT1-MMP was increased. CONCLUSION: HSC is sensitive to the oxygen, hypoxia enhances the expression of MMP-2 and the effect is more marked at the early stage; hyperoxia mainly raises the activity of MMP-2.

关键词： Animals Cell Division Cell Hypoxia Cells, Cultured Gelatinase A Gene Expression Regulation, Enzymologic Hepatocytes Hyperoxia Metalloendopeptidases RNA, Messenger Rats Rats, Sprague-Dawley Research Support, Non-U.S. Gov't tissue inhibitor of metalloproteinase-2

Effect of a nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester on invasion of human colorectal cancer cell line SL-174T

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World Journal of Gastroenterology 2005年第40期11卷 6385-6388页

作者： Li-Bo YU Xin-Shu Dong Wen-Zhou sun Dong-Lu Zhao Yue Yang Department of GeneralSurgery the Affiliated Tumor Hospital of Harbin MedicalUniversity Harbin 150040 Heilongiiang Province China Cancer Research Institute of HarbinMedical University Harbin 150040 Heilongjiang Province China

AIML To investigate the effect and mechanism of action of the nitric oxide synthase （NOS） inhibitor NG-nitro-L-arginine methyl ester （L-NAME） on invasion and metastasis of human colorectal cancer cell line SL-174T. METHODS： Human colorectal cancer cel4 line SL-174T was cultured and treated separately with four different dosages of L-NAME for 72 h, Nitric oxide （NO） production was measured with Griess reagent, The effect of L-NAME on invasion and migration of SL-174T cells were evaluated by using Transwell chambers attached with polycarbonate filters and reconstituted basement membrane （Matrigel）, RT-PCR was performed to determine the mRNA levels of matrix metalloproteinase-2 （MMP-2） and tissue inhibitor metalloproteinase-2 （TIMP-2）,RESULTS： L-NAME could significantly inhibit NO production of SL-174T in a dose-dependent manner. After being treated for 72 h with 0.2, 0.4, 0.8, and 1.0 mmol/L L- NAME, respectively, the ability of the L-NAME treated SL- 174T cells to invade the reconstituted basement membrane decreased significantly （t = 8.056, P〈0.05; t= 14.467, P〈0.01; t= 27.785, P〈0.01; and t= 29.405, P〈0.01, respectively） and the inhibition rates were 10.29%, 19.62%, 34.08%, and 42.23%, respectively. Moreover, L-NAME could inhibit migration of SL-174T cells, and the inhibition rates were 20.76%, 24.95%, 39.43%, and 46. 85% for L-NAME at 0.2, 0.4, 0.8, and 1.0 mmol/L, respectively （t = 15.116, P〈0.01）. In addition, after treatment with L-NAME, expression of MMP-2 mRNA was significantly decreased （t = 71.238, P〈0.01） and that of TIMP-2 mRNA was markedly increased （t = -13.020, P〈0.01）. CONCLUSION： L-NAME exerts anti-invasive and anti- metastatic effects on SL-174T cell line via downregulating MNP-2 mRNA expression and upregulating TIMP-2 mRNA expression.

关键词： Colorectal Neoplasms Neoplasm Invasiveness Cell Line, Tumor Cell Movement Dose-Response Relationship, Drug Female Gelatinase A Humans Male NG-Nitroarginine Methyl Ester Nitric Oxide Nitric Oxide Synthase inhibitors tissue inhibitor of metalloproteinase-2

Methodologic research on TIMP-1,TIMP-2 detection as a new diagnostic index for hepatic fibrosis and its significance

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World Journal of Gastroenterology 2002年第2期8卷 282-287页

作者： Oing-He Nie Yong-Oian Cheng Yu-Mei Xie Yong-Xing Zhou Bai-Xian Guang Yi-Zhan Cao,The Centre of Diagnosis and Treatment for Infectious Disease of Chinese PLA,Tangdu Hospital,Fourth Military Medical University,Xi’an 710038,Shanxi Province,China The Centre of Infectious Disease Diagnosis and Treatment of Chinese PLA Tangdu Hospital Fourth Military Medical University Xi'an 710038 Shaanxi Province China. nieqinghe@***

AIM: To set up a new method to detect tissue inhibitors of metalloproteinase-1 and -2(TIMP-1 and TIMP-2) in sera of patients with hepatic cirrhosis, and to investigate the expression and location of TIMP-1 and TIMP-2 in liver tissue of patients with hepatic cirrhosis, and the correlation between TIMPs in liver and those in sera so as to discuss whether TIMPs can be used as a diagnosis index of hepatic fibrosis. METHODS: The monoclonal antibodies (McAbs) of TIMP-1 and TIMP-2 were used to sensitize erythrocytes, and solid-phase absorption to sensitized erythrocytes (SPASE) was used to detect TIMP-1 and TIMP-2 in the sera of patients with hepatic cirrhosis. Meanwhile, with the method of in situ hybridization and immunohistochemistry, we studied the mRNA expression and antigen location of TIMP-1 and TIMP-2 in the livers of 40 hepatic cirrhosis patients with pathologic diagnosis. RESULTS: With SPASE, they were 16.4% higher in the acute hepatitis group, 33.3% higher in the chronic hepatitis group, and the positive rates were 73.6% and 61.2% respectively in sera of hepatic cirrhosis patients, which were remarkably higher than those in chronic hepatitis and acute hepatitis group (Ptissues, all of them showed positive expression of TIMP-1 and TIMP-2 mRNA detected with immunohistochemistry or in situ hybridization (positive rate was 100%). Expression of TIMPs in different degrees could be found in liver tissue with cirrhosis. TIMPs were located in cytoplasm of liver cells of patients with hepatic cirrhosis. There was a significant correlation between serum TIMPs level and liver TIMPs level. CONCLUSION: SPASE is a useful method to detect the TIMP-1 and TIMP-2 in sera of patients with hepatic cirrhosis, and TIMP-1 and TIMP-2 can be considered as a useful diagnostic index of hepatic fibrosis, especially TIMP-1.

关键词： Antibodies, Monoclonal Erythrocytes Humans Immunoassay In Situ Hybridization Liver Liver Cirrhosis Protease inhibitors Research Support, Non-U.S. Gov't tissue inhibitor of metalloproteinase-1 tissue inhibitor of metalloproteinase-2

Inhibition of Metastatic Progression of SSTR2 Gene Transfection Mediated by Adenovirus in Human Pancreatic Carcinoma Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2006年第1期26卷 68-71页

作者：冯延平黄涛高军常青秦仁义 Department of Pancreatic-biliary Surgery Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China

The inhibition of metastatic progression of Somatostatin receptor type 2 （SSTR2） gene transfection mediated by adenovirus in human pancreatic carcinoma cells and the mechanisms involved in this effect were studied. The full-length human SSTR2 cDNA was introduced into the pancreatic cancer cell line BXPC-3 by adenovirus-mediated transfection. Stable expression of mRNAs and protein of SSTR2 was detected by RT-PCR and Western-blot. The Matrigel-coated Transwell was used to detect the migratory and invasive ability of SSTR2-expressing cells, Adv-GFP control cells and mock control cells. Furthermore, the expression of matrix metalloproteinase-2 （MMP-2） and tissue inhibitor of metalloproteinase-2 （TIMP-2） was detected by RT-PCR in these cells. The stable expression of SSTR2 was detected in BXPC-3 transfected by Adv-GFP-SSTR2. A dramatic decrease of BXPC-3 expressing sst2 cells migrating through a Matrigel-coated filter was observed, as compared with Adv-GFP control and mock control cells （P〈0. 01）. Moreover, the expression of MMP-2 mRNA was significantly reduced in the SSTR2-expressing cells and converse- ly the expression of TIMP-2 mRNA was significantly increased in the SSTR2-expressing cells when compared with the Adv-GFP control and mock control （P〈0. 01）. The expression of reintroduced human SSTR2 gene in BXPC-3 cells by Adv-GFP-SSTR2 had the anti-migratory and anti-invasive effects, and the mechanisms involved in this effect may be due to the down-regulated expression of MMP-2 and up-regulated expression of TIMP-2.

关键词： pancreatic carcinoma adenovirus vector somatostatin receptor type 2, metalloproteinase-2 tissue inhibitor of metalloproteinase-2