| Background:Breast cancer,which plays a major role in the incidence of malignant tumors,is one of the most prevalent cancers in female worldwide.With the development of economy and the change of lifestyle,the incidence of breast cancer is increasing year by year.The number of new cases and deaths of breast cancer in our country accounted for 12.2%and 9.6%of the world.About 90%of breast cancer patients died from distant metastases.Therefore,exploring the mechanism of distant metastasis and finding an effective molecular target of preventing of distant metastasis to improve the prognosis of patients with breast cancer is extremely important.As a non-steroidal anti-inflammatory drug,aspirin was used for antipyretic analgesia at first,but nowadays the purpose is mainly to prevent thrombosis and other diseases.In 1968,Gasic first proposed the potential role of aspirin in tumor prevention and treatment,and suggested that aspirin could prevent distant metastasis.Subsequent animal experiments,epidemiological investigation and clinical trials had confirmed that aspirin had a significant impact on the inhibiting the development of breast cancer,gastrointestinal cancer,prostate cancer,liver cancer and other neoplastic diseases,but its specific mechanism has not been clarified.Previous studies had found that the increased platelets could affect the prognosis of the tumors,meanwhile,activated platelets could secrete a variety of cytokines to make the tumor cells epithelial-mesenchymal transition through interacting with tumor cells,and thus increasing the ability of invasion and migration.Objective:To identify the effect of aspirin on epithelial-mesenchymal transition,migration and invasion in MCF-7 cells induced by activated platelets.Methods:1.Effects of aspirin on proliferation of MCF-7 cells(1)CCK-8 was used to determined the effects of different concentrations of aspirin on proliferation of MCF-7 cells.2.Effects of aspirin,activated platelets which were treated with arachidonic acid and different concentrations of aspirin pretreatment of arachidonic acid respectively on the epithelial mesenchymal transition in MCF-7 cells.Experimental grouping:a.control group:An equal volume of 1640 medium was added.b.low dose aspirin group:Adding the final concentration of 0.5mmol/1 aspirin.c.high dose aspirin group:Adding the final concentration of 2mmol/aspirin.d.platelet group:Adding the final concentration of arachidonic acid 20mmol/l,the final concentration of platelets was 150000/1.e.platelet group with low dose aspirin:0.5 mmol/1 aspirin,20 mmol/1 arachidonic acid and platelets were added.f.platelet group with high dose aspirin:Adding 2mmol/l aspirin,20mmol/l arachidonic acid and platelets.(2)The morphological changes of tumor cells were observed under inverted microscope according to the experimental grouping method.(3)Expression of E-cadherin and Vimentin,tumor cell epithelial mesenchymal transformation marker proteins,were detected by Western Blot.3.Effects of aspirin,activated platelets which were treated with arachidonic acid and different concentrations of aspirin pretreatment of arachidonic acid respectively on the ability of migration and invasion in MCF-7 cells.Results:1.0.5mmol/l-2mmol/l aspirin had no effect on the proliferation of MCF-7 cells,and the proliferation of MCF-7 cells was significantly suppressed when the concentration of aspirin was more than 5mmol/l.2.Activated platelets were co-cultured with MCF-7 cells could induce EMT,and adding activated platelets which were treated with aspirin pretreatment of arachidonic acid,aspirin could inhibit EMT in MCF-7 cells were mediated by platelets.3.Effects of aspirin,activated platelets and platelets were treated with aspirin on the migration and invasion ability in MCF-7 cells.(1)Cell scratch test showed that the closed score of the platelet group was significantly higher than the control group,meanwhile,the score of the platelet group with low and high dose aspirin were reduced obviously than the platelet group,but no significant difference compared with the control group.The migration and invasion capacity of MCF-7 cells were detected by transwell.The results suggested that the migration and invasion ability of the platelet group were significantly enhanced than the control group.The ability of migration and invasion in the platelet group with low and high dose aspirin were decreased than the platelet group,but no significant difference compared with the control group.Conclusion:Activated platelets were co-cultured with MCF-7 cells could induce epithelial-mesenchymal transition,and enhance the ability of migration and invasion.Aspirin could reduce platelet activation through suppressing cyclooxygenase activity and inhibit epithelial-mesenchymal transition,migration and invasion ability in MCF-7 cells induced by activated platelets. |
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