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Abstract Background: Qualea grandiflora (QG) (Vochysiaceae), also known as "pau-ferro", "pau-terra" or "pau-de-tucano", is a very common deciduous tree in the Brazilian Cerrado used in traditional medicine to treat inflammations, ulcers, diarrhea, and infections. There are reports in the scientific literature that demonstrate the medicinal effects of the bark and leaf of the QG. However, studies involving this plant are rather imited. Aim of the study: To perform the phytochemical analysis of the QG hydroalcoholic extract (HAE) of leaves, and to investigate it effects on fibroblast and preosteoblasts. Methods: Phytochemical analysis was done by HPLC-DAD. Murine NIH/3T3 fibroblasts and MC3T3-E1 preosteoblasts cell lines (ATCC) were used for the experiments. Cell viability was assessed by the MTT colorimetric assay and the expression of MMP-14 and HIF-1α by immunofluorescence. Results and conclusion: The following compounds were identified by HPLC-DAD, such as quinic acid, ethyl galate, ellagic acid derivatives as O-methylellagic acid O-galloyl, O-methylellagic acid O-deoxyhexoside, galloyl derivatives, flavonol glycoside as kaempferol-O-deoxyhexoside, quercetin-O-deoxyhexoside, myricetin-O-deoxyhexoside and the pentacyclic triterpene arjunglucoside. Cell viability results demonstrated no cytotoxic effects in the studied concentrations. We found in QG HAE some compounds with therapeutic properties that can increase the expression of MMP-14 and HIF-1α, in fibroblasts and preosteoblasts. These data suggest that QG HAE has an action on these two molecules widely involved in physiological conditions, such as collagen remodeling, bone development and growth and pathological processes as HIF signaling in cancer metastasis.
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Objective:Study on the inhibitory effect of celecoxib on pancreatic cancer SW1990 cells growth.Methods:The experiment was divided into two parts.(1) SW1990 cells were divided into fluorouracil group (50 μ g/mL),celecoxib group(50 μ mol/L),the combined group (celecoxib 50 μ mol/L+fluorouracil 50 μ g/mL),blank group,after 24 h culture of each group using the MTT assay SW1990 cell growth inhibition rate was detected by flow cytometry cell cycle distribution ratio;(2) were used at different concentrations (12.5,25.0,50.0 and 100.0 μ mol/L) of celecoxib,while using different concentrations above celecoxib were combined with 50 μ g/mL of fluorouracil treated SW1990 cells were treated 24 h,using real-time fluorescence polymerase chain reaction(RT-PCR) to detect differential expression Survivn RNA and detect the cyclooxygenase-2 (COX-2) enzyme-linked immunosorbent assay(ELISA),matrix metalloproteinase-14(MMP-14) expression differences.Results:Celecoxib group,fluorouracil group was significantly higher than the rate of the combination group cytostatic blank group (P＜0.05),the inhibition rate of the combination group was significantly higher than the celecoxib group and fluorouracil group (P＜0.05),celecoxib cloth inhibition rate was significantly lower than the fluorouracil group(P＜0.05);significantly higher than the proportion of cells celecoxib group,fluorouracil group,joint group of G0/G1 phase control group (P＜0.05),celecoxib group was significantly higher than the proportion of cells fluorouracil group G0/G1 phase combination group(P＜0.05),significantly higher than the proportion of cells fluorouracil group G0/G1 phase celecoxib group (P＜0.05);with the increasing concentration of celecoxib,celecoxib group,celecoxib+Survivn RNA fluorouracil group,COX-2 and MMP-14 protein levels were decreased and group differences were between statistically significant(P＜0.05),Under the condition of the same concentration of celecoxib and fluorouracil plus celecoxib group RNA survivin,COX-2 and MMP-14 protein expression levels were significantly lower than that of celecoxib group,the difference has statistical significance (P＜0.05).Conclusion:Celecoxib on the growth of pancreatic cancer SW1990 cells was significantly inhibited in a dose dependent manner,and its mechanism may be related to reduced Survivn RNA,COX-2 and MMP-14 protein expression.
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Objective To preliminarily explore the effects of human microRNA-181a on migration of gastric cancer cells and its mechanism.Methods The expression of miRNA-181a-5p in gastric cancer cell line GC9811 and peritoneal high metastasis gastric cancer cell line GC9811-P were tested by quantitative real-time polymerase chain reaction (qRT-PCR ). GC9811 cell line was transfected byendogenous synthetic analog mimic and its homologous negative control of miRNA-181a-5p,which were considered as up-regulated group and its control group respectively;GC9811-P were transfected by miRNA inhibitor and its homologous negative control of miRNA-181a-5p,which were considered as down-regulated group and its control group respectively.After miRNA-181a-5p was up or down-regulated,cell proliferation,migration and apoptosis capabilities of gastric cells were detected by matrix thiazolyl tetrazollium (MTT)assay,cloning-forming assay,Transwell migration test,wound healing assays and apoptosis test.After miRNA-181a-5p was up or down regulated,the changes of matrix metalloproteinase (MMP)14 expression were determined by Western blot.Independent sample t test was performed for mean comparison between samples and chi square test was used for rate comparison.Results The results of qRT-PCR showed the relative expression quantity of miRNA-181a-5p in GC9811 was 1 .000 00 ± 0.021 26 and in GC9811-P was 3.175 61 ±0.106 76,and the difference was statistically significant (t =34.620,P <0.01 ).The results of MTT assay indicated that the cell proliferation rate of up-regulated group was higher than that of up-regulated control group,and that of down-regulated group was lower than down-regulated control group.The cloning-forming assay demonstrated that the number of clone forming and clone forming rate of up-regulated group,up-regulated control group,down-regulated group and down-regulated control group were 234.00±10.12 and 46.8%,93.00±9.61 and 18.6%,51 .00 ±7.96 and 10.2%,99.00±8.05 and 19.8%,respectively.The differences between up-regulated group,down-regulated group and their control group were statistically significant (t = 17.500,7.344,χ2 = 12.27, 9.51 ,all P <0.01).The results of Transwell migration experiment showed the number of cells migrated through membrane hole of up-regulated group was 164.00±19.31 ,and which was higher than that of up-regulated control group (87.00±23.04,t=4.436,P <0.05);that of down-regulated group was 157.00± 11 .50,and which was lower than that of down-regulated control group (234.00 ±12.12,t =7.982,P <0.05).The result of wound healing assays indicated that the rate of migration distance of up-regulated group,up-regulated control group,down-regulated group and down-regulated control group were 2.09 ± 0.18,1 .27 ±0.23,1 .15 ±0.15 and 1 .67 ±0.12,respectively.The differences between up-regulated group,down-regulated group and their control group were statistically significant (t =4.863 and 4.689, both P <0.05).The results of apoptosis experiments demonstrated that the apoptosis rate of up-regulated group,up-regulated control group,down-regulated group and down-regulated control group were (6.10± 1 .02 )%,(9.10 ± 2.13 )%,(12.70 ± 1 .23 )%,(8.70 ± 2.54 )%,respectively,and there was no statistically significant difference between up-regulated group,down-regulated group and their control group (both P *0.05).The results of Western blot showed that the grey value of up-regulated group was 561 .881 ±35 .740,which was higher than that of up-regulated control group (275 .784±23.520);that of down-regulated group was 579.565 ±37.950,which was lower than that of down-regulated control group (1 312.760±51 .270),and the differnces were statistically significant (t =11 .580 and 19.910,both P <0.01).Conclusion miRNA-181a-5p highly expresses in peritoneal high metastasis gastric cancer cell line GC9811-P and promoted the proliferation and migration of gastric cancer cell line GC9811 and GC9811-P with a tendency to suppress apoptosis.
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Objective To explore the expression of matrix metalloproteinase-14(MMP-14)protein in the human stomach carcinoma tissues and its correlation with carcinoma invasiveness and metastasis.Methods The MMP-14 protein expression was detected by immunohistochemistry in 59 cases of stomach carcinoma tissues (observation group)and 20 cases of normal stomach tissues (control group,the adjacent normal tissues from the tumor margin of 5 cm confirmed by pathology),and its correlation with the clinically pathological parameters was analyzed.The expression characteristics of MMP-14 among various TNM stages of stom-ach carcinoma were also analyzed.Results The positive rate of MMP-14 expression was 50.85%(30/59)in the observation group and 5.00% (1/20)in the control group,the positive rate of the observation group was significantly higher than that of the control group (P <0.01);the expression level of MMP-14 was correlated with the differentiation degree,regional lymph node metastasis degree,invasion depth,lymphatic invasion and TNM stage,which showing the statistical difference(P < 0.01);the expression of MMP-14 protein was up-regulated and showed the transferring trend from cytoplasm to cellular membrane along with the progres-sion of TNM stage.Conclusion The overexpression of MMP-14 protein exists in stomach carcinoma tissues,which contributing to the invasion and metastasis of stomach carcinoma cells.
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Objective To investigate the expression characteristics of Glypican3 ,MMP-9 and MMP-14 in primary hepatocellular carcinoma ,and to focus on their roles on the development ,progress and metastasis of tumor .Methods 102 cases of primary hepato-cellular carcinoma were taken as the observation group and 80 cases of normal liver tissues as the control group .The expressions of Glypican3 ,MMP-9 and MMP-14 were detected by the immunohistochemistry method and their expression difference in different clinicopathological characteristics and the correlation were investigated .Results The positive rates of Glypican3 ,MMP-9 and MMP-14 expressions in the observation group were significantly higher than those in the control group .The positive rates of Glypi-can3 ,MMP-9 and MMP-14 expressions were closely correlated with the tumor volume ,differentiation degree ,lymph node metasta-sis ,vessel infiltration ,membrane invasion and PCNA expression .The correlation analysis showed that the positive relationships were found between Glypican3 and MMP-9 ,between Glypican3 and MMP-14 and between MMP-9 and MMP-14 in the observation group(r=0 .48 ,P=0 .024 1 ;r=0 .46 ,P=0 .013 2;r=0 .43 .P=0 .031 3) .The survival analysis showed that expressions of Glypi-can3 ,MMP-9 and MMP-14 were correlated with the patient′s prognosis .Conclusion The higher-expressions of Glypican3 ,MMP-9 and MMP-14 may promote the occurrence and development of primary hepatocellular carcinoma .Detecting the postoperative expres-sions of Glypican3 ,MMP-9 and MMP-14 has certqain value to judge the prognosis in primary hepatocellular carcinoma .
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Objective To evaluate the expression and clinical significance of matrix metalloproteinase 14(MMP-14) in osteoar-thritis(OA) patients in synovium and synovial fluid of knee joint .Methods Semi-quantitative RT-PCR were used to detect the ex-pression of MMP-14 mRNA in 32 case of OA (experimental group)and 26 non-OA patients(control group)synovium ;protein level expression of MMP-14 in synovial fluid were analyzed by ELISA .Results The expression of MMP-14 mRNA in experimental group synovium of knee joint was significantly increased compared with control group (P<0 .05);but the protein level of MMP-14 was low in experimental group synovial fluid compared with control group (P<0 .05) .Conclusion The expression of MMP-14 is increased in OA patients synovium and maybe promote the progress of OA directly ;the protein level of MMP-14 is decreased in OA synovial fluid ,indicated the function of MMP-14 in OA synoviocyte indirectly .
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Objective To investigate the correlation between the expression levels of semaphorin3A and MMP14,and their subsequent prognostic significance in nonsmall cell lung cancer(NSCLC).Methods The expression of semaphorin3A and MMP14 protein levels was analyzed in 94 cases of NSCLC tissues and in 80 cases of normal lung tissues,using immunohistochemistry(IHC).Correlation and survival analysis were used to further investigate their association and prognostic value.The correlation analysis using Pearson test,and log-rank test for survival analysis.Results The NSCLC tissues exhibited a lower expression of semaphorin3A and a higher expression of MMP14 than in the control lung tissues.The downregulation of semaphorin3A and upregulation of MMP14 may promote pleural invasion,lymph node metastasis,vascular invasion and proliferating cell nuclear antigen expression.The expression of semaphorin3A was correlated with the maximum diameter of tumor.There was a negative correlation between the protein expression levels of semaphorin3A and MMP14 in NSCLC tissues.Conclusion The data suggest that lower expression of semaphorin3A and a higher expression of MMP14 may promote occurrence and development in NSCLC and that the combined detection of semaphorin3A and MMP14 protein may be a helpful tool in predicting the prognosis of NSCLC.
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PURPOSE: The purpose of this study was to compare and quantify the expression of C-reactive protein (CRP), matrix metalloproteinase (MMP)-14, and tissue inhibitor of metalloproteinases (TIMP)-2 in gingival tissues of patients with chronic periodontitis accompanied with inflammatory reaction related to alveolar bone resorption with or without type 2 diabetes mellitus (DM). METHODS: Twelve patients with type 2 DM and chronic periodontitis (group 3), twelve patients with chronic periodontitis (group 2), and twelve healthy individuals (group 1) were included in the study. Gingival tissue biopsies were collected from each patient and from healthy individuals at the time of periodontal surgery (including surgical crown lengthening) or tooth extraction. The concentrations of cytokines were determined by a western blot analysis. RESULTS: The expression levels of CRP and MMP-14 increased in group 2 and 3, and they were highest in group 3. The expressions of TIMP-2 also increased in group 2 and 3. CONCLUSIONS: This study demonstrated that the expression levels of CRP, MMP-14, and TIMP-2 might be inflammatory markers in periodontal inflamed tissue. It can be assumed that CRP, MMP-14, and TIMP-2 may be partly involved in the progression of periodontal inflammation associated to type 2 DM.
Subject(s)
Humans , Biopsy , Blotting, Western , Bone Resorption , C-Reactive Protein , Chronic Periodontitis , Crowns , Cytokines , Diabetes Mellitus, Type 2 , Inflammation , Matrix Metalloproteinase 14 , Tissue Inhibitor of Metalloproteinase-2 , Tissue Inhibitor of Metalloproteinases , Tooth ExtractionABSTRACT
BACKGROUND: CD44 is a cell surface receptor that has been implicated in tumor cell invasion and metastasis in a range of tumors of various organs, including breast, ovary, colon, lung, and brain. CD44 stimulates the invasive ability by interacting with matrix metalloproteinase 14 (MMP14). The expression of MMP14 on the cell surface is thought to trigger multiple proteinase cascades and to stimulate cell migration. METHODS: A total 54 astrocytoma patients were eligible for this study. We performed a retrospective clinicopathological review and CD44 and MMP14 immunohistochemistry. RESULTS: The expressions of CD44 and MMP14 were significantly correlated with the World Health Organization (WHO) grade. On univariate analysis, the WHO grade and the expression of CD44 were the significant prognostic factors affecting overall survival (OS) and disease progression free survival (DPFS). On the multivariate analysis by the Cox regression model, the only WHO grade was shown to be a significant independent prognostic factor for predicting the DPFS and OS. CONCLUSIONS: In this study, the CD44 and MMP14 expressions were related to the WHO grade of astrocytoma. The CD44 expression status was a prognostic factor for DPFS and OS on univariate analysis, but it was not an independent prognostic factor on the multivariate analysis.