Adenocarcinoma, Follicular ; genetics ; metabolism ; pathology ; Biomarkers, Tumor ; genetics ; Diagnosis, Differential ; Humans ; Matrix Metalloproteinase 1 ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; Thyroid Neoplasms ; genetics ; metabolism ; pathology ; ras Proteins ; biosynthesis ; genetics

OBJECTIVETo investigate the effects of E-cadherin on the biologic behavior of SUM149, an inflammatory breast cancer cell line.

METHODSSUM149 cells were transfected with dominant-negative mutant E-cadherin expressing plasmid. The positive clones with higher expression of dominant-negative E-cadherin mutant were identified by RT-PCR and fluorescent flow cytometry method. The differences in cell growth, proliferation and invasion between positive clones and controls were compared.

RESULTSWhereas the proliferation of positive clones was of no change, compared with controls, the ability of invasion was decreased and the mRNA levels of MMP-1 and MMP-9 were downregulated. Gelatin zymography analysis also confirmed the decreasing expression of MMP-9 in the positive clones.

CONCLUSIONIn this cell line model, down-regulation of E-cadherin can inhibit the ability of invasion of this inflammatory breast cancer cell line.

Breast Neoplasms ; metabolism ; pathology ; Cadherins ; biosynthesis ; genetics ; physiology ; Cell Line, Tumor ; Down-Regulation ; Female ; Humans ; Matrix Metalloproteinase 1 ; biosynthesis ; genetics ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Mutation ; Neoplasm Invasiveness ; Plasmids ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

This paper is aimed to present a research on fusion protein of human tumor necrosis factor-alpha (hTNF-alpha), matrix metalloproteinase 1 (MMP1), and foldon sequence using the methord of gene engineering. We transformed the recombinant plasmid, which contains the DNA sequences of hTNF-alpha, MMP1, and foldon sequence, into Rosetta2, and successfully induced the fusion protein to express under given conditions by isopropyl beta-D-1-Thiogalactopyranoside (IPTG). Then we purified the expression product through a glutathione S-transferase (GST) resin and collected the interested protein. This research may lay the groundwork for scientific research and clinical application.
Base Sequence ; Escherichia coli ; genetics ; metabolism ; Humans ; Matrix Metalloproteinase 1 ; biosynthesis ; genetics ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

OBJECTIVETo compare the effects of matrix metalloproteinase (MMP) inhibitor doxycycline, losartan, and their combination on the expression of MMP-8, 13, tissue inhibitor of MMP-1, 2 (TIMP-1, 2), and collagen remodeling in the noninfarcted myocardium after acute myocardial infarction (AMI) in rats.

METHODSTwo hundred and fifty-four AMI rats, induced by left coronary ligation, were randomly assigned to the following groups: (1) AMI controls group (n = 64); (2) doxycycline group (30 mg x kg(-1) x d(-1), n = 63); (3) losartan group (10 mg x kg(-1) x d(-1), n = 62); (4) concomitant doxycycline and losartan group (30 and 10 mg x kg(-1) x d(-1) respectively, n = 65); and (5) Sham-operated rats (n = 30), which were randomly selected to serve as noninfarction controls. Each group was further divided into three subgroups of 1, 2, and 4 weeks that received treatment. After the completion of treatment, the rats were killed. The mRNA and protein expression of MMPs and TIMPs in the noninfarcted myocardium were quantified by RT-PCR and Western blot, respectively. The type I and type III collagen volume fraction (CVF) of the noninfarced myocardium were assessed immunohistochemically.

RESULTSNo significant difference existed in myocardial infarction sizes among the 12 subgroups of AMI controls and the three treatment groups (42%-48%, all P > 0.05). Compared with sham operated rats, the mRNA and protein expression of MMP-8 and 13 significantly increased by 39%-183% in all three subgroups of AMI controls (all P < 0.05), except both of their mRNA expressions in 2-week subgroups; the mRNA and protein levels of TIMP-1 increased only in 1-week subgroup of AMI controls by 104% and 67%, respectively (both P < 0.05); the mRNA of TIMP-2 increased in all 1, 2, and 4-week subgroups by 144%-232% (all P < 0.05), but its protein expression lagged and only enhanced in 2 and 4-week subgroups of AMI controls by 231% and 332%, respectively (both P < 0.05). Meanwhile, both type I and type III CVF of noninfarcted myocardium significantly increased in all three subgroups of AMI controls (type I CVF: 3.01%-5.64% vs 1.53%-1.67%, P < 0.01-0.001; type III CVF: 2.19%-4.42% vs 1.46%-1.59%, P < 0.05-0.001), with type I CVF being higher in 4-week than in 1 and 2-week subgroups (5.64% vs 3.01% and 3.02% respectively, all P < 0.05). Compared with AMI controls, all three kinds of treatment significantly reduced the increased mRNA and protein expressions of MMP-8, 13 and TIMP-1, 2 after AMI by 14%-60% (all P < 0.05), as well as type I/III CVF in their 2 and 4-week subgroups (type I CVF: 1.56%-2.38% vs 3.02%-5.64%, P < 0.05-0.001; type III CVF: 1.92%-2.65% vs 4.19%-4.42%, P < 0.05-0.01), except for doxycycline's effect on type III CVF in any of its three subgroups (all P > 0.05). Among the three treatment groups, significant differences existed in the above mentioned indicators only at some subgroup levels (all P < 0.05).

CONCLUSIONSLike losartan, doxycycline can also suppress the enhanced mRNA and protein expression of MMP-8, 13 and TIMP-1, 2, and reduce type I collagen deposition in the noninfarcted myocardium after AMI in rats. However, it has no effect on type III collagen deposition.

Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Animals ; Collagen Type I ; biosynthesis ; genetics ; Collagenases ; biosynthesis ; genetics ; Doxycycline ; pharmacology ; Drug Synergism ; Female ; Losartan ; pharmacology ; Matrix Metalloproteinase 13 ; Matrix Metalloproteinase 8 ; biosynthesis ; genetics ; Matrix Metalloproteinase Inhibitors ; Myocardial Infarction ; metabolism ; Myocardium ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; Tissue Inhibitor of Metalloproteinase-2 ; biosynthesis ; genetics ; Tissue Inhibitor of Metalloproteinases ; biosynthesis ; genetics

Actins ; biosynthesis ; genetics ; Adult ; Aged ; Female ; Hepatitis B, Chronic ; complications ; Humans ; Liver Cirrhosis ; blood ; enzymology ; virology ; Male ; Matrix Metalloproteinase 1 ; biosynthesis ; genetics ; Middle Aged ; Plasminogen Activator Inhibitor 1 ; blood ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; Urokinase-Type Plasminogen Activator ; blood

Animals ; Liver Cirrhosis ; enzymology ; Male ; Matrix Metalloproteinases, Secreted ; biosynthesis ; genetics ; Mice ; RNA, Messenger ; biosynthesis ; genetics ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics

In situ hybridization and immunocytochemical techniques were employed to examine the expression of matrix metalloproteinases-1 (MMP-1) and to identify the pattern of its distribution in rat pancreas. The results indicated that the signal of MMP-1 mRNA and MMP-1 positive immunoreaction were detected in some fiberoblasts around interlobular ducts and exocrine cell in margin acinus of some lobules, but the signal of MMP-1 mRNA and MMP-1 positive immunoreaction could not be detected in most of other acinus and islets of pancreas. It is concluded that the expression of MMP-1 in above cells of rat might play an important role in acinar proliferation and differentiation of rat pancreatic tissues.
Animals ; Cell Division ; Immunohistochemistry ; In Situ Hybridization ; Male ; Matrix Metalloproteinase 1 ; analysis ; biosynthesis ; genetics ; Pancreas ; enzymology ; RNA, Messenger ; analysis ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of Yangjing Zhongyu decoction (YZD) on metalloproteinase-9 (MMP-9) and its inhibitor-1 (TIMP-1) expression and sex hormone regulation in mid-luteal phase endometrium of women with cryptogenic infertility.

METHODSIn situ hybridization and reverse transcription-polymerase chain reaction method was used to detect MMP-9 and TIMP-1 mRNA, and radioimmunoassay was used to determine levels of serum estradiol (E2) and progesterone (P) synchronously, of 22 infertile women during mid-luteal phase.

RESULTSAfter treatment, the mid-luteal serum E2 and P level was 451.501 +/- 226.342 pmol/L and 46.502 +/- 19.948 nmol/L respectively, significantly higher than that before treatment (304.656 +/- 135.853 pmol/L and 33.782 +/- 15.459 nmol/L respectively), the difference was significant (P < 0.01). Staining of MMP-9 mRNA positive granules in cytoplasm and nuclei of adeno-epithelial cell mid-luteal phase endometrium deepened significantly, but the change in mesenchym was insignificant. The MMP-9 mRNA expression after treatment was 0.617 +/- 0.186 (grey level), significantly higher than the level before treatment (0.490 +/- 0.370), comparison between them showed significant difference (P < 0.05). Change of TIMP-1 mRNA expression in adeno-epithelial and mesenchym before and after treatment was insignificant (0.588 +/- 0.191 vs 0.621 +/- 0.146, P < 0.05). Correlation analysis showed that the quantitative difference of P value before and after treatment was positively correlated with the difference of MMP-9 mRNA before and after treatment (r = 0.682, P < 0.01).

CONCLUSIONYZD could soothen Gan and nourish Shen, raise the level of mid-luteal phase serum P, and further promote MMP-9 gene expression in endometrium to benefit the degradation of extracellular matrix of endometrium, and facilitate for blastocyst implantation.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Endometrium ; metabolism ; Estradiol ; blood ; Female ; Humans ; Infertility, Female ; drug therapy ; metabolism ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Phytotherapy ; Progesterone ; blood ; RNA, Messenger ; biosynthesis ; genetics ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics

OBJECTIVETo investigate the influence of baicalin on the IL-1beta induced pro-MMP-1 in HGF and the effects of baicalin on MMP-3 expression in periodontal ligament cells (PDLCs).

METHODSThe amount of secreted pro-MMP-1 and MMP-3 expression was detected by ELISA and cell immunochemistry.

RESULTS(1) The amount of secreted pro-MMP-1 (3.333 +/- 0.123) microg/L increased significantly following 1 microg/L of IL-1beta, compared with control group (1.960 +/- 0.180) microg/L. Addition of baicalin to cell culture medium for 1 hour following IL-1beta decreased pro-MMP-1 secretion in a dose-dependent manner in the range of 10 approximately 1,000 microg/L. (2) 1 microg/L IL-1beta could significantly stimulate the synthesis and secretion of MMP-3 in PDLCs. (3) The baicalin could not interfere the synthesis of MMP-3, but could inhibit the release of MMP-3 from PDLCs.

CONCLUSIONSBaicalin could inhibit the secretion of pro-MMP-1 and MMP-3 expression in IL-1beta induced HGF and PDLCs, which suggests that baicalin may play an important role in preventing and treating periodontal disease.

Collagenases ; biosynthesis ; genetics ; Enzyme Precursors ; biosynthesis ; genetics ; Fibroblasts ; enzymology ; pathology ; Flavonoids ; pharmacology ; Gingiva ; enzymology ; pathology ; Humans ; Interleukin-1 ; pharmacology ; Interleukin-1beta ; Matrix Metalloproteinase 1 ; Metalloendopeptidases ; biosynthesis ; genetics ; Peptide Fragments ; pharmacology ; Periodontal Ligament ; enzymology ; pathology ; Periodontitis ; enzymology ; pathology ; Scutellaria ; chemistry

OBJECTIVETo study the effect of laminarin polysaccharide (LP) on the activity of matrix metalloproteinase of photoaging skins.

METHODKunming SPF mice were prepared with back hair shaved, and randomly divided into the control group, the model group, the LP low does group (LP-L, 1 mg x kg(-1)), the LP high dose group (LP-H, 5 mg x kg(-1)) and the Vit E (100 mg x kg(-1)) group. They were abdominally injected with drugs twice on a daily basis. Except for the control group, all groups were exposed to ultraviolet rays for 1 hour every day, five times on a weekly basis, with accumulated exposure dose of UVB being 21.60 J x cm(-2) and accumulated exposure dose of UVA being 84.02 J x cm(-2). Eight weeks later, exposed back skins were collected to detect thickness of dermis by HE stain, content of hydroxyproline (Hyp) by chemical colorimetry, and serum MMP-1 and TIMP-1 content by ELISA. In addition, matrix metalloproteinase-1 (MMP-1) mRNA and relative content of tissue inhibitor of metalloproteinase-1 (TIMP1) mRNA was analyzed with Real-time PCR.

RESULTCompared with the model group, the LP-H group could significantly increase the thickness of dermis, skin Hyp content and serum TIMP-1 level, and decrease relative content of MMP-1 mRNA in skin and MMP-1 content in serum.

CONCLUSIONLP can regulate the metabolism of collagen photoaging skins by adjusting the activity of matrix metalloproteinase.

Animals ; Female ; Glucans ; Matrix Metalloproteinase 13 ; biosynthesis ; genetics ; metabolism ; Mice ; Plant Extracts ; chemistry ; pharmacology ; Plants, Medicinal ; chemistry ; Polysaccharides ; chemistry ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Skin Aging ; drug effects ; physiology ; radiation effects ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; genetics ; metabolism ; Ultraviolet Rays