High Expression Of MTA2 In Ovarian Epithelial Cancer: Correlation With Tumorigenesis And Cancer Progression

Objectives:The aim of this study is to assess MTA2 expression and its clinical significance in ovarian epithelial cancer.MethodsThe MTA2 levels in 4 ovarian cell lines were analyzed using semi-quantitative RT-PCR and Western-blot. MTA2 expression was assessed in normal, borderline, benign, malignant epithelial ovarian tissues by using immunohistochemical technique.ResultsThe MTA2 expression level was significant higher in high invasive ovarian cell lines SW626 and A27 than that in the cell lines with low invasive potential, CAOV3 and OVCAR3 cells(P<0.01). Compared to normal and begin tissues, borderline ovarian tissues expressed higher level MTA2 (p<0.01), and the highest expression level of MTA2 was observed in malignant ovarian tissues (p<0.01). The increased expression of MTA2 was associated with aggressive behaviors of epithelial ovarian cancer according to the fact that the expression levels of MTA2 were correlated with late clinical stage, high histopathological grade and lymphoid metastasis.ConclusionsThis study provides the first evidence that MTA2 may play an important role in the tumorgenesis and progression of epithelial ovarian cancer. The overexpression of MTA2 may be a biomarker in progression of epithelial ovarian cancer. Objective:The aim of this study is to investigate the mechanism that MTA2 participates the progression of the epithelial ovarian cancer.Method:MTA2 and E-Cadherin protein expressions in 47 fresh frozen ovarian cancer specimens were detected by Western-blot. The antisense MTA2 eukaryotic expression vector(MTAas) was constructed and transfected into ovarian cancer A2780 by lipofectamine. After G418 selection for 4 weeks, the cell line with stable expression of the gene was obtained. Then the expression level of E-Cadherin and MTA2 were measured by western-blot. Cell proliferation was evaluated by MTT method. The anchorage-independent growth was assessed by soft agar assay. Wound healing assay was performed to evaluate the cell migration.Results:Samples from total 27 ovarian cancer patients were divided into two groups by E-Cadherin expression level-detectable (positive) or undetectable (negative). E-Cadherin-negative tumors presented higher level MTA2 expression than that of E-Cadherin-positive tumors (P<0.001). Antisense RNA reduced 81.4% expression of MTA2 protein, while increased the E-Cadherin expression from undetectable to detectable level in A2780 cell. The downregulation of MTA2 inhibited cancer cell migration and reduced soft agar colony formation. In addition, the transfection of antisense MTA2 inhibited the cell proliferation of the A2780 cell.Conclusion:MTA2 overexpression contributes the aggressive phenotypes of the ovarian cancer cell possibly through the downregulation of E-Cadherin. Objective:The aim of the current study was to assess the value of extensive surgical staging including pelvic and para-aortic lymphadenectomy and the impact of the adjuctive therapies, the hormone therapy and the radiotherapy, on patients with stageⅠendometrial cancer(EC).Methods:In the retrospective trial,67 patients were enrolled including 37 stageⅠpatients,8 stage patients, 14 stageⅢpatients and 8 stage IV patients. Patients in stageⅠwere divided into three groupsⅡbased on the surgical operation underwent, including subradical/radical hysterectomy salpingo-oophorectomy plus retroperitoneal lymphadenectomy or sampling(group A), subradical/radical hysterectomy salpingo-oophorectomy (group B),hysterectomy salpingo-oophorectomy plus retroperitoneal lymphadenectomy(group C).The overall 5-years survival rate were evaluated respectively. The benefits and side effects of the adjuvant therapies on the prognosis of stageⅠpatients were assessed. The recurrence site and the cause of death were discussed.Result:The 5-years survival rates of patients with EC by clinical stage were as follows:stageⅠ88.65%,stageⅡ86.67%,stageⅢ77.92% and stageⅣ28.57% with a overall survival rate of 78.99%. Grade 3 histology, adenosquamous or clear cells cancer, and more than one third invasion of the muscle or involving in lymphnodes related to low survival rate and were, therefore, defined as high risk factors for EC. In stageⅠ, patients with high risk factors had 20% lower 5-year survival rate than patients without high risk factors. Three surgical strategies represented the extent of operation did not affect the patients survival. However,12.28%(7/57) clinical stageⅠpatients were proved to be stageⅢby group A surgery. Hormone therapy significantly increased the 5-year survival rate of stageⅠpatients. In contrast, the radiotherapy had no effect on stageⅠpatients' survival. The recurrence rates of stageⅠ, stageⅡand stageⅢwere 2.7%(1/37),12.5%(1/8) and 7.2%(1/14), respectively. Recurrent sites were 60% inside and 40% outside the pelvic. The death rates of EC were StageⅠ0.81%%(4/37), StageⅡ12.5%(1/8), StageⅢ21.43%(3/14) and StageⅣ50%(4/8). The causes of death were 84.62%(11/13) of cancer specific death, 7.69%(1/13)of radiotheraputic ileuses and 7.69%(1/13) of cardiovascular diseases. The hormone therapy had fewer side effects than other therapies.Conclusion:The enlargement of operation field and retroperitoneal lymphadenectomy or sampling provided the accurate surgical stage but no therapeutical benefits for StageⅠpatients. The adjuvant therapies should be given to StageⅠpatients after operation due to the improvement of StageⅠpatients' prognosis by hormone therapy. The radiotherapy needs further evaluation by increasing the population size of the study. ADAMTS13 cleaves von Willebrand factor (vWF) between Tyr1605 and Met1606 residues at the central A2 subunit. The aminoterminus of AD AMTS 13 protease appears to be sufficient to bind and cleave vWF under static and denatured condition. However, the role of the carboxylterminus of AD AMTS 13 in substrate recognition remains controversial. Present study demonstrates that ADAMTS13 cleaves vWF in a rotation speed- and protease concentration-dependent manner on a mini vortexer. Removal of the CUB domains (delCUB) or truncation after the spacer domain (MDTCS) significantly impairs its ability to cleave vWF under the same condition. ADAMTS13 and delCUB (but not MDTCS) bind vWF under flow with dissociation constants (Kd) of about 50 nM and about 274 nM, respectively. The isolated CUB domains are neither sufficient to bind vWF detectably nor capable of inhibiting proteolytic cleavage of vWF by AD AMTS 13 under flow. Addition of the TSP1 5-8 (T5-8CUB) or TSP1 2-8 repeats (T2-8CUB) to the CUB domains restores the binding affinity toward vWF and the inhibitory effect on cleavage of vWF by ADAMTS13 under flow. These data demonstrate directly and quantitatively that the cooperative activity between the middle carboxyl-terminal TSP1 repeats and the distal carboxylterminal CUB domains may be crucial for recognition and cleavage of vWF under flow. Objective:The aim of this study is to demonstrate that bivalirudin acts as an anti-platelet agent in PCI patients by preventing activation of PARs on the platelet surface.Methods:Total 22 patients undergoing elective percutaneous coronary intervention (PCI) were recruited into the study. Bivalirudin plasma levels at 6 sequential time point were measured by LC/MS/SM method. Platelet aggregation in platelet rich plasma was performed at baseline and 30 min after the initiation of bivalirudin or the end of PCI if the procedure was completed earlier than 30 min in patients' platelet rich plasma (PRP) or normal healthy donors'PRP spiked with bivalirudin. PAR1 expression levels on the platelet surface was evaluated at the same time point with aggregation assay by detecting the binding of the antibody Span 12 that only recognizes intact PAR1 on Flow Cytometry System.Results:The stable plasma level of bivalirudin was were 2.7±0.5μM which correlated with marked inhibition of thrombin-induced platelet aggregation and significantly inhibited cleavage of PAR1, but not affected platelet aggregation in response to PAR1 specific agonist, SFLLRN, and PAR4 agonist, AYPGKF. Bivalirudin markedly right-shifted the thrombin aggregation curves by 26-fold to a mean thrombin EC50 of 256±87 nM in the patients undergoing PCI. Bivalirudin displayed a linear concentration dependent inhibition of thrombin-induced platelet aggregation if spiked into the healthy donors' PRP. The thrombin-platelet aggregometry dose-response curve of bivalirudin determined for a range of bivalirudin concentrations in whole blood gave a predicted mean plasma bivalirudin level of 3.1μM which deviated by only 13% from the actual measured mean steady-state value of 2.7μM in the patient samples. Termination of the bivalirudin infusion resulted in rapid clearance of the drug with a half-life of 29.3 minutes.Conclusions:Bivalirudin effectively suppresses thrombin-dependent platelet activation via inhibition of PAR1 cleavage. Platelet aggregation might be potentially used to evaluate the effective concentration of bivalirudin and platelet status. The short half life of bivalirudin contributes its less bleeding in patients undergoing PCI. Background:The aim of this study is to evaluate the effect of bivalirudin on platelet procoagulant activity and thrombin generation in patients undergoing percutaneous intervention (PCI).Methods:Total 22 patients undergoing PCI were recruited into the study. Plasma thrombin-antithrombinⅢ(TAT) levels of patients at 3 sequential time points(before,30 min after initiation of bivalirudin or the end of PCI if the procedure was completed within 30 min and 2h after the stop of bivalirudin) were measured by ELISA. Aggregations in platelet rich plasma (PRP) from patients at first two time point or from healthy donors were performed. Platelet procoagulant activity was determined by detecting the binding of AnnexivⅤby flow cytometry. Thrombin generation in PRP from healthy donors was measured by using its substrate S2238 in vitro.Results:Bivalirudin infusion reduced systemic thrombin levels by more than 50% during PCI. Unexpectedly, bivalirudin also significantly inhibited collagen-platelet aggregation by 11%. Collagen induced a conversion of the platelet surface to a procoagulant state in a thrombin-dependent manner which was blocked by bivalirudin infusion in patients undergoing PCI or by being spiked into healthy donors' PRP. Bivalirudin inhibited in vitro thrombin generation in PRP caused by typeⅠcollagen.Conclusions:Bivalirudin effectively inhibits collagen-induced platelet procoagulant activity as well as systemic thrombin levels in patients undergoing PCI.