Objective To examine the effects of newly designed LY52 on the expression of matrix metalloproteinases and invasive ability of hepatocellular carcinoma HepG2 cells. Methods The effects of LY52 on the proliferations of HepG2 cells were detected by MTT assay. Gelatin zymography and Western blot were used to detect the effects of LY52 on matrix metalloproteinase-2 expression in the cell line. Transwell chamber assay was used to detect the effects of LY52 on the invasion of the cells. Results No obvious inhibitory or cytotoxicity effects of LY52 was found in lower concentrations (lt;200 μg/ml) of LY52. Gelatin zymography and Western blot showed that matrix metalloproteinase-2 expression were inhibited by LY52 in a dose-dependent manner in HepG2 cells. Furthermore, transwell chamber assay showed that LY52 could significantly inhibit the invasion of the cell line in a dose-dependent manner.Conclusion The results suggest that LY52 may inhibit the invasion of hepatocellular carcinoma cells by suppressing the matrix metalloproteinase-2 activity.
Objective To observe the effect of epidermal growth factor (EGF) on the proliferation, adhesion, invasiveness and the activation of nuclear factor-κB (NF-κB), matrix metalloproteinases (MMPs) expression and explore related mechanisms in pancreatic cancer cells. Methods Cell invasion assay, proliferation assay and adhesion assay were used to examine the proliferation, adhesion and invasiveness of pancreatic cancer cells, respectively. NF-κB activity was detected by electrophoretic mobility shift assay (EMSA), and MMPs protein and mRNA expressions were investigated by gelatin zymography, Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Results EGF increased the invasiveness of pancreatic cancer cell in a dose-dependent manner (P＜0.05), but did not affect cell proliferation or adhesion. The expressions of MMP-9 mRNA and protein significantly increased after induction by EGF and were highest when EGF concentration was 50 ng/ml, while there was no effect on the expressions of MMP-2 mRNA and protein. Furthermore, NF-κB activity increased with increased concentration of EGF in a concentration-dependent manner (P＜0.05). In addition, NF-κB activity and the expressions of MMP-9 mRNA and protein by pretreatment with both pyrrolidine dithiocarbamate (PDTC) and EGF decreased when compared that by pretreatment with EGF alone. The invasiveness of pancreatic cancer cell by pretreatment with both PDTC and EGF decreased when compared that by pretreatment with EGF alone and nothing (P＜0.05).Conclusion The findings indicate that the NF-κB-mediated MMP-9 induction is essential for EGF-induced invasiveness in pancreatic cancer cells, which can be inhibited by PDTC.
ObjectiveTo explore the inhibition action of valproic acid to inflammatory cells and smooth muscle cells then to find out that valproic acid (VPA) can repress rat thoracic aortic aneurysm or not.
MethodsThe model of rat thoracic aortic aneurysm was built through the method of soaking the adventitia of artery using porcine pancreatic elastase (PPE). The rats were divided into three groups:a normal saline blank control group (a C group), an adventitia soaked PPE group (a P group), and adventitia soaked PPE plus intraperitoneal injection by injecting intraperitioneal VPA 200 mg/kg for seven days (a PV group).The animals of the three groups were all using vascular ultrasound to detect blood vessel diameter. Animals were killed after operation to observe the general morphology of vascular aneuysm and do the immunohistochemial, morphological, protein analysis of interleukin 1 (IL-1), interleukin 6 (IL-6), smooth muscle 22 alpha (SM22α), matrix metallopeptidase 2 (MMP-2), MMP-9 and Western blot by drawing animals on the 14th day.
ResultsThe vessels diameter in the PV group was narrower than that in the P group (P value<0.05). HE staining, immunohistochemistry and Western blot displayed that the cells in the P group were in disorder arrangement and interstitial disorder while the cells in the PV group maintained better albumin layer. The protein expressions of IL-1, IL-6, MMP-2, and MMP-9 in the PV group decreased except that SM22α increased.
ConclusionVPA can inhibit phenothpic transforming of aneurysm inflammatory cells and smooth muscle cells, reduce the levels of cell proliferation, decrease the secretion of matrix metalloproteinases, and depress tumor growth of rat thoracic aorta.
ObjectiveTo summarize the biological function of extracellular matrix metalloproteinase inducer (EMMPRIN) in tumor progression, and its roles in clinical diagnosis and treatment in recent years.
MethodsLiteratures about the recent studies on molecular structure of EMMPRIN and biological function in tumor progression were reviewed according to the results searched from PubMed database.
ResultsEMMPRIN play important roles in the tumor progression, involved in inducing the degradation of extracellula matrix, promoting angiogenesis, inhibiting apoptosis, enhancing chemoresistance and so on.
ConclusionEMMPRIN could be a potential therapeutic target in turmor.
Purpose
To identify matrix metalloproteinase (MMP) in human vitreous samples of diabetic vitreoretinopathy (DR) and other ocular diseases (non-DR) and to probe the related factors of MMP expression．
Methods
Thirty-one diabetic and 17 non-diabetic vitreous samples (nine macular hole and eight epiretinal membrane patients) were examined． Samples were concentrated and subjected to substrate zymography to conduct a quantitative analysis of MMP-2,9 activity． The technology of Western blotting against anti-human MMP-2,9 was performed to identify MMP in vitreous samples．
Results
Vitreous samples both from DR patients and from non-DR patients showed a single band at the position of 72 kDa, correspondin g to MMP-2． Quantitative analysis revealed that diabetic vitreous showed higher MMP-2 activity than non-DR, although the difference was not significant.45.2% of DR patients showed MMP-9, but no expression in non-DR．Among DR samples, the positive ratio of MMP-9 in partial posterior vitreous detachment (PVD)(66．7%) was significantly higher than that of complete PVD (15．4%)． Western blotting study confirmed the expression of MMP-2 and MMP-9．
Conclusion
There is no obvious difference of MMP-2 activity between DR and non-DR． MMP-9 may be involved in the pathogenesis of diabetic vitreor etinopathy and the deterioration of proliferative change．
(Chin J Ocul Fundus Dis, 2001,17:195-197
Objective
To investigate the expression of E26 transformation-specific-1（E26ts-1），matrix metalloproteinase-1（MMP-1）and tissue inhibitor of matrix metalloproteinase-1 （TIMP-1） in choroidal melanoma and the correlation with the tumorprime;s infiltration and metastasis.
Methods
Immunohistochemistry was used to detect the expression of E26ts-1，MMP-1 and TIMP-1 in 78 cases of choroidal melanoma who were divided into shuttle-cells，paraepithelial-cells and mixed-cells type according to the configuration of tumor cells.The patients were followed up and their average existing time was calculated.The results were statistically computed with statistic SPSS 10.0 package.
Results
In the 78 cases，shuttle-cells type was found in 21，paraepithelial-cells type in 34，and mixed-cells type in 23. Expression of TIMP-1was low in uveal melanoma，while expression of E26ts-1 and MMP-1 was obviously found in the three types of choroidal melanoma;the sequence of expression intensity was shuttle-cells，mixed-cells and paraepithelial-cells type.Among 37 cases who had been followed up，the shuttle-cells type was in 18 with the average existing time of （78.33plusmn;24.69）months，the mixed-cells type was in 10 with the average existing time of（61.44plusmn;20.46）months，and the paraepithelial-cells type was in 9 with the average existing time of（36.76plusmn;12.19）months.The existing time was negative correlated with the intensity of expresion of E26ts-1 and MMP-1.
Conclusion
The high expression of E26ts-1 and MMP-1and low expression of TIMP-1may relate to the choroidal melanomaprime;s infiltration and metastasis.
(Chin J Ocul Fundus Dis, 2006, 22: 174-176)
Objective To investigate the feasibil ity of alendronate (ALN) in treating osteoarthritis (OA) by observing the effects of ALN on interleukin 1β (IL-1β) induced chondrocytes of rat in vitro. Methods The chondrocytes of knee articular surface from 15 SD rats (1-month-old, female or male, weighing 100-150 g) were cultured. The chondrocytes were observed by inverted phase contrast microscope and identified by toluidine blue staining and HE staining. The third passage chondrocytes were divided into 3 groups: the chondrocytes were cultured with DMEM for 5 days in group A, with 10 ng/mL IL-1β for 2 days and with DMEM for 3 days in group B, and with 10 ng/mL IL-1β for 2 days and with 1 × 10-6 mol/L ALN for 3 days in group C. Immunocytochemistry and real-time PCR were performed to determine the expression levels of collagen type II (Col II), matrix metalloproteinase 13 (MMP-13), and β-catenin. Results Toluidine blue staining proved that the cultured cells were chondrocytes. The integrated absorbency (IA) value of Col II in group C (10.290 7 ± 0.499 2) was lower than that in group A (15.377 0 ± 0.571 8) and higher than that in group B (5.463 2 ± 0.450 4), showing significant differences (P lt; 0.05). The IA value of MMP-13 in group C (3.068 6 ± 0.205 6) was significantly lower than that in group B (6.998 1 ± 0.329 7, P lt; 0.05), but there was no significant differenc when compared with group A (2.777 5 ± 0.199 6, P gt; 0.05). The IA value of β-catenin in group C (6.611 7 ± 0.381 8) was lower than that in group B (11.799 9 ± 0.348 7) and higher than that in group A (4.390 3 ± 0.551 9), showing significant differences (P lt; 0.05). The mRNA expression of Col II in group C was significantly higher than those in groups A and B (P lt; 0.05), the mRNA expression of MMP-13 in group C was significantly lower than that in group B (P lt; 0.05) but there was no significant difference when compared with group A (P gt; 0.05). The mRNA expression of β-catenin in group C was significantly lower than that in group B (P lt; 0.05) and higher than that in group A (P lt; 0.05). Conclusion ALN can protect rat chondrocyte from OA induced by IL-1β in vitro possibly by upregulating Col II and inhibiting the expression of MMP-13 and β-catenin in the chondrocytes.
Objective To explore the expressions of E-cadherin， matrix metalloproteinases-2 （MMP-2） protein，and matrix metalloproteinases-9 （MMP-9） protein in gastric cancer tissues， and to analyze the possible statistical relati-onship between the expressions of E-cadherin， MMP-2 protein， and MMP-9 protein， and clinicopathological features ofgastric cancer. Methods The ABC immunohistochemical staining was adopted to examine the expressions of E-cadherin，MMP-2 protein， and MMP-9 protein in 40 paraffin slices of gastric cancer （gastric cancer group）， with the adjacent tissue as the control group （adjacent tissue group）. The positive rates of 3 kinds of protein were compared between the2 groups， in addition， the statistical relationship between the expressions of the 3 kinds of protein and clinicopathological features of gastric cancer was examined respectively by SPSS 19.0 software. Results The expressions of E-cadherin， MMP-2 protein， and MMP-9 protein were all found in gastric cancer tissues and adjacent tissues. In gastric cancer tissue group， the expression of E-cadherin downregulated while the expressions of MMP-2 protein and MMP-9 protein upregulated in comparison to adjacent tissue group （P＜0.05）. The significant association was found between the expre-ssion of E-cadherin and the gastric cancer tissues of T3＋T4 stage， N1-N3 stage， and Ⅲ＋Ⅳ stage， which had lower positive expression rate （P＜0.05）. The expression of MMP-2 protein in gastric cancer tissues of M1 stage and Ⅲ＋Ⅳstage upregulated （P＜0.05）， and the expression of MMP-9 protein upregulated in gastric cancer tissues of T3＋T4 stage，Ⅲ＋Ⅳ stage， or lowlydifferentiated＋undifferentiated （P＜0.05）. No significant relationship was found in other clinical-pathological features and 3 kinds of protein except aforementioned significant relationship （P＞0.05）. Conclusions In the development progress of gastric cancer， the E-cadherin may get involved in the mechanism of tumor invasion and lymph node metastasis， MMP-2 protein may get involved in the mechanism of distant metastasis， and MMP-9 protein may get involved in the mechanism of differentiation and tumor invasion. The examination of those 3 kinds of markers may play an role in the judgment of tumor stage and estimation of prognosis in gastric cancer clinically.
Objective To observe the expression of matrix metalloproteinase（MMP-2, MMP-9 and vascular endothelial growth factor (VEGF) in retinoblastoma (RB) and its relationship with the differentiation and optic nerve infiltration of RB.Methods Forty paraffin specimens of pathological confirmed RB were studied. They were divided into differentiated group (15 cases) and undifferentiated group (25 cases) , optic nerve infiltration group（13 cases） and without optic nerve infiltration group（27 cases）. The expression of MMP-2, MMP-9 and VEGF were detected by immunohistochemistry, their relationships with the differentiation and optic nerve infiltration were also analyzed.Results The positive rate of MMP-2, MMP-9 and VEGF expression in 40 RB cases were 52.5％，57.5％ and 72.5% respectively．The expression of MMP-2, MMP-9 and VEGF in the undifferentiated group were significantly higher than those in the differentiated group (chi;2=9.037, 9.253, 8.095; P＜0.05). The expression of MMP-2, MMP-9 and VEGF in RB with optic nerve infiltration group were significantly higher than those in RB without optic nerve infiltration group (chi;2=11.045,10.243, 8.956；P＜0.05). The expression of MMP-2，MMP-9 had a positive correlation with the expression of VEGF in RB (r=0.126,0.314；P＜0.05). Conclusions MMP-2, MMP-9 and VEGF expressed in RB tumor tissues. The expression of MMP-2, MMP-9 has a positive correlation with the expression of VEGF. The levels of MMP-2, MMP-9 and VEGF expression are related to optic nerve infiltration of RB cells.
Objective
To detect the variation rule of different cellular components, extracellular matrix, matrix-metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases（TIMPs）in proliferative membranes in proliferative vitreoretinopathy (PVR) with different courses of disease, and to investigate the remodeling mechanism of PVR.
Methods
Sixteen surgically excised specimens of proliferative membranes from patients with rhegmatogenous retinal detachment combined with PVR with the course of disease of 2 months to 8 years were selected. The different cellular component of retinal pigment epithelial (RPE) cells and glial cells, component of extracellular matrix including fibronectin, laminin，and collagen types Ⅰ to Ⅳ, and matrix metalloproteinases (MMP2, MMP9) and TIMP1 in proliferative membrane were labeled by immunohistochemical method. The variati on of those labeled components in proliferative membrane in PVR duration and the correlation between these components and the course of PVR were analyzed.
Results
As the duration of PVR increased,the expression of RPE cells, fibronectin and MMP2 decreased (Plt;0.05）,while glial cells,collagen type Ⅰ and Ⅲ increased (Plt;0.05）.The positive staining of laminin and collagen type Ⅱ and Ⅳ were found, but the association with PVR duration was not detected. A negative correlation between PVR duration and RPE cells, MMP2, and fibronectin respectively and a positive correlation between PVR duration and glial cells, collagen Ⅰand Ⅲ respectively were detected. MMP2 positively related with variation of fibronect in. Positive staining of MMP9 and TIMP1 was recorded but did not change with the variation of the disease course.
Conclusion
During the formation and development of proliferative membrane in PVR, RPE cells, glial cells, fibronectin, collagen type Ⅰand Ⅲ and MMP2 take part in the remodeling of proliferative membrane.
(Chin J Ocul Fungdus Dis, 2006, 22:308-312)
