Objective To review the progress of a disintegrin and metalloproteinase with thrombospondin motif 4 (ADAMTS-4) and ADAMTS-5 in osteoarthritis. Methods Recent literature about the ADAMTS-4 and -5 in osteoarthritis was analyzed; the structure, function, inhibitors of the ADAMTS-4 and -5, and the relationship between the proteases and osteoarthritis were analyzed and summarized. Results ADAMTS-4 and -5 can reduce chondrocyte and extracellular matrix by degrading aggrecan and cartilage oligomeric matrix protein, which induced the pathogenesis of osteoarthritis. Conclusion ADAMTS-4 and -5 have been demonstrated to play important roles in osteoarthritis. It can better guide treatment and prevention of osteoarthritis to further study related mechanism of ADAMTS-4 and -5, and to promote the establishment of a clinical drug targets.
ObjectiveTo investigate the effect of ADAM33 gene silencing in VSMCs on the proliferation and lumen formation of airway vascular endothelial cells (VECs) in a co-culture system and the possible regulatory mechanism. MethodsThe Human aortic smooth muscle cells (HASMCs) and human pulmonary microvascular endothelial cells (HPMECs) were used to construct a cell co-culture system. ADAM33 gene expression was silenced by lentivirus transfection technique, and the subjects were divided into endothelial cell blank group, co-culture group, co-culture +shRNA negative control group, and co-culture + ADAM33-SHRNA group. The expressions of sADAM33, VEGFA,VEGER2, ang-1 and ang-2 in co-culture system were detected by ELISA. The proliferation and lumen formation of HPMECs were observed by CCK-8 and Transwell experiments. The protein expression of Tie2, PI3K, Akt, and mTOR key molecules in Tie2/PI3K/Akt/mTOR signaling pathway and the phosphorylation levels of AKT and mTOR were detected by Western-blotting method. Results① Compared with the co-culture group (0.851±0.036) and the co-culture + shRNA negative control group (0.828±0.047), the OD value of the co-culture + ADAM33shRNA group (0.699±0.038) was significantly decreased (P<0.05). ② Compared with the co-culture group (159.169±15.740) and the co-culture +shRNA negative control group (157.357±21.612), the tube length of the co-culture +ADAM33shRNA group (120.812±2.791) was also significantly decreased (P<0.05). ③ After ADAM33 gene expression of HASMCs was silted in co-culture system, the expression levels of VEGFA, VEGFR2, ang-1 and ang-2 were significantly decreased (P<0.05), while the expression levels of Tie2, PI3K, P-Akt and P-mtor were decreased (P<0.05). ConclusionsSilencing the expression of the ADAM33 gene could reduce the release of sADAM33 from the membrane of the airway VSMCs, regulate the proliferation and lumen formation of airway VECs by reducing the expression of VEGF/VEGFR and inhibiting the activities of the Tie2/PI3K/Akt/mTOR signaling pathways,and then participate in airway vascular remodeling in asthma.
ObjectiveTo investigate the association between polymorphism of V4,F+1 in ADAM33 (adisintegrin and metalloproteinase 33) gene and COPD in a northwestern Uighur population. MethodsA total of 100 Uighur COPD patients and 140 healthy volunteers were recruited in the study. Genotypes were determined by restriction fragment lengthpolymorphism(PCR-RFLP). All subjects had a epidemiological investigation including modified british medical research council(mMRC),COPD assessment test(CAT),and pulmonary function test. The 100 Uighur COPD patients were assessed by revised GOLD2011. ResultsAssessed by revised GOLD2011,the patients of A,B and C grade accounted for 22%,35% and 30%,respectively. There was no statistical significance in the distributions of the V4,F+1 alleles between the patients and the controls(P>0.05). There was no statistical significance between SNPs in ADAM33(V4 and F+1) with the decreased lung function and the grade of COPD(P>0.05). ConclusionThere was no association between polymorphism of V4,F+1 in ADAM33 gene and COPD in a northwestern Uighur population.
Objective To study relationship between integrins and carcinogenesis, development, treatment or prognosis of gastric cancer. Methods The literatures about integrins and gastric cancer in recent years were reviewed and analyzed. Results The current study found that the β1 subunit integrins and αν subline integrins are closely associated with the gastric cancer. The β1 subunit integrins are associated with the invasion and metastasis of the gastric cancer, the αν subline integrins are associated with the typing, grading, and staging of the gastric cancer, and the ανβ3, ανβ5 and ανβ6 are associated with the prognosis of the gastric cancer, further more, the ανβ6 could be used as an independent effective prognostic factor. Conclusions Integrins are associated with occurrence, development, treatment, and prognosis of gastric cancer. It′s mechanism such as signal transduction pathway is not completely clarified. With further in-depth research, it′s molecular mechanism would be gradually elucidated and provide new ideas and methods for diagnosis, treatment, and prognosis of gastric cancer.
Objective To investigate the role of calcium- and integrin-binding protein-1（CIB1） in oxidized lowdensity lipoprotein（OX-LDL） inhibiting migration of mouse macrophages. Methods To silence CIB1 express of mouse macrophages by RNA interference, then incubating mouse macrophages with OX-LDL, cell migration and cell spreading of mouse macrophages were analyzed. Results At 24-72h after macrophages transfected CIB1 siRNA, the express of CIB1 protein was restrained obviously. To silence CIB1 express could increase migration and spreading of mouse macrophages significantly. Conclusions CIB1 plays the important role in intracellular modulating mechanism of OX-LDL inhibiting mouse macrophages migration.
Objective To investigate the expression of ADAM9 in breast cancer and its clinical significance. Methods The expressions of ADAM9 in normal breast tissues and breast cancer tissues were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry, and whose relationship with clinicopathologic features was analyzed. Results The expression of ADAM9 mRNA increased in the breast cancer tissues, but which was not detected in the normal breast tissues. The expression of ADAM9 protein in the breast cancer tissues was significantly higher than that in the normal breast tissues (Plt;0.05), and which in the metastatic lymph nodes was significantly higher than that in the negative lymph nodes or corresponding primary lesions (Plt;0.05). The expression of ADAM9 in the breast cancer tissues was correlated with the lymph node metastasis and histological grade (Plt;0.05). Conclusion ADAM9 is overexpressed in the breast cancer tissues, which might involve in the pathological progression of breast cancer.
OBJECTIVE: To investigate the selection and identification of human keratinocyte stem cells(KSC) in vitro. METHODS: According to the characteristics of KSC which can adhere to extracellular matrix very fast, we selected 3 groups of different time(5 minutes, 20 minutes and 60 minutes) and unselected as control group. And the cells were identified by monoclone antibody of beta 1-integrin and cytokeratin 19 (Ck19), then the image analysis was done. Furthermore we analyzed the cultured cells with flow cytometer(FCM) and observed the ultrastructure of the cell by transmission electron microscope(TEM). RESULTS: The cell clones formed in all groups after 10 to 14 days, while the cells of 5 minute group grew more slowly than those of the other groups, however, the clones of this group were bigger. The expression of beta 1-integrin and Ck19 were found in all groups. The positive rate of beta 1-integrin was significant difference between 5 minute group and the other groups (P lt; 0.05). And the expression of Ck19 was no significant difference between 5 minute group and 20 minute group(P gt; 0.05), and between 60 minute group and control group. But significant difference was observed between the former and the later groups(P lt; 0.05). The result of FCM showed that most cells of the 5 minute group lied in G1 period of cell cycle, which was different from those of the other groups. At the same time, the cells of 5 minute group were smaller and contained fewer organelles than those of the other groups. CONCLUSION: The above results demonstrate that the cells of 5 minute group have a slow cell cycle, characteristics of immaturity, and behaving like clonogenic cells in vitro. The cells have the general anticipated properties for KSC. So the KSC can be selected by rapid attachment to extracellular matrix and identified by monoclone antibody of beta 1-integrin and Ck19.
ObjectiveTo investigate the expression of a disintegrin and metalloproteinase with thrombospondin typeⅠmotif (ADAMTS1) in colorectal cancer tissues, and to study the relationship with clinicopathological features and prognosis of it. MethodsExpression of ADAMTS1 was evaluated by immunohistochemistry (SP method) in 65 specimens, which obtained by resection from patients with colorectal cancer, including corresponding adjacent benign tissues. Chi-square test was used for analyzing the relationship between expression of ADAMTS1 and clinicopathological features of colorectal cancer tissues. Cox proportional hazard model was used to explore the relationship between expression of ADAMTS1, other clinicopathological parameters, and patients' survival situation. ResultsThe positive expression rate of ADAMTS1 was 40% (26/65) in the colorectal cancer tissues and 85% (55/65) in the adjacent benign tissues, which was significantly higher in adjacent benign tissues (χ2=27.546, P < 0.001). The positive expression rate of ADAMTS1 was significantly lower in the colorectal cancer tissues with lymph node metastasis than that of the colorectal cancer without lymph node metastasis (χ2=5.329, P=0.021). Results of survival analysis showed that median survival time were 27 months in the ADAMTS1-negative group and 70 months in the ADAMTS1-positive group respectively, and the survival situation was better in latter group (χ2=10.151, P=0.001). Results of multivariable prognostic analysis of Cox proportional hazard model showed that colorectal cancer withⅠ-Ⅱstage (RR=3.782, 95% CI:1.509-9.476, P=0.005), without lymph node metastasis (RR=3.107, 95% CI:1.186-8.138, P=0.021), and with positive-expression of ADAMTS1 (RR=2.020, 95% CI:1.071-3.809, P=0.030) had better survival situation. ConclusionsExpression of ADAMTS1 is down-regulated in colorectal cancer tissues and it is associated with lymph node metastasis. The prognosis of patients in ADAMTS1-positive group is better than that of ADAMTS1-negative group, suggesting that ADAMTS1 may be an independent prognostic factor in colorectal cancer.
ObjectiveTo summarize the relationship between integrins, tumor metabolism, and tumor cells with pancreatic stellate cells in the tumor microenvironment, in order to provide targets and ideas for the treatment of pancreatic ductal adenocarcinoma.MethodTo review the literatures on pancreatic stellate cells, integrins, and amino acid metabolism as therapeutic targets for pancreatic ductal adenocarcinoma in the domestic and overseas.ResultsThe drug research for pancreatic ductal adenocarcinoma was currently under vigorous development, but remain in the animal and clinical test stage. As a new therapeutic protein, ProAgio could inhibit the expression of integrin αvβ3, activation and secretion of pancreatic stellate cells, and alanine metabolism in the microenvironment of pancreatic ductal adenocarcinoma, so as to achieve the dual effects of anti-fibrosis and anti-tumor.ConclusionsThe roles of activated pancreatic stellate cells, ProAgio, integrin αvβ3, and alanine metabolism in pancreatic ductal adenocarcinoma have been partially elucidated, but the specific mechanism still needs further investigation and may become a completely new therapeutic target someday.
ObjectiveTo investigate the relationship between DDX46 genes and invasion and migration of esophageal squamous cell carcinoma cells. MethodsHuman esophageal squamous cell carcinoma cells TE-1 were transfected by fluorescent marker shRNA lentivirus (shDDX46 group), and an empty vector was transfected as a control (shCtrl group). The expression rate of green fluorescent protein under the microscope was used to evaluate the cell transfection efficiency. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blotting (WB) detected the knockdown efficiency of the target gene at the mRNA and protein expression levels. Wound healing, invasion assay and migration assay detected the changes of invasion and metastasis ability. Classical pathway analysis was used to explore signaling pathway changes and the possible mechanism of DDX46 in the invasion and metastasis was explored by detecting fibronectin expression. ResultsDDX46 gene at mRNA and protein levels was significantly inhibited after lentiviral transfection. Wound healing showed that after 8 h the cell mobility of TE-1 cells decreased significantly (P=0.001). Invasion assay showed that after 24 h the average cell metastasis rate of TE-1 cells was lower in the shDDX46 group than that in the shCtrl group (P<0.001). The cell metastasis rate in the shDDX46 group corresponding to observation points in the transwell assay was lower than that in the shCtrl group (P<0.001) after 24 h culture. The results of the classical pathway analysis showed that the integrin signaling pathway activity was inhibited, further exploration of the mechanism of action found that the expression of fibronectin associated with cell adhesion was decreased. ConclusionDDX46 gene is related to the invasion and migration ability of esophageal squamous cell carcinoma cells. Knockdown of DDX46 genes may reduce cell adhesion by downregulating the integrin pathway signaling.