The Interaction Between Cx43,AKAP95 And Its Associated Proteins Affects Cell Cycle Progression

Lung cancer ranks first in the incidence and death of malignant tumor in China,and its incidence is still rising in recent years.The incidence and mortality of breast cancer are high in many cities in China,which has become the threat to women's health.Traditional chemotherapy has been used as an effective treatment for cancer,but its efficiency is low and its toxicity are serious.It has reached the plateau stage.Molecular targeted therapy is a new type of cancer treatment.Finding valuable molecular markers is very important for molecular targeted therapy.In this study,the proteins related to the progression of the lung cancer and breast cancer were studied,and the proteins related to the cell cycle process were further studied at the cellular level.The first part is the immunohistochemistry experiment.Objective:To assess the levels of exchange protein directly activated by cAMP(Epac1)and protein kinase C(PKC)in lung cancer and para-carcinoma tissues.In addition,correlation between the two proteins was assessed,and the associations of Epacl and PKC levels,respectively,with A-kinase anchor protein 95(AKAP95)and Connexin43(Cx43)amounts were examined.In breast cancer tissues,to investigate Epacl,phosphodiesterase 4(PDE4)and PKC expression,and the correlation of the aforementioned proteins expression level.In addition,the correlation of the expression of Epacl,PDE4 and PKC proteins with AKAP95 and Cx43 proteins was investigated.Methods:Protein expression levels were detected by the PV-9000 Two-step immunohistochemistry technique.Conclusion:1.Epacl protein may be involved in the incidence,development and metastasis of lung cancer.But in breast cancer tissue,Epacl protein may promote the progression of cancer,suggesting that the protein may have tissue specificity in the regulation of cancer process;2.PDE4 protein may promote the progression of breast cancer;3.In lung cancer tissue,the expression of Epacl and PKC protein,Epacl and Cx43 protein,and PKC and AKAP95 protein expression in lung cancer tissues are associated.It is suggested that Epacl may regulate the process of lung cancer cell cycle by regulating PKC and Cx43 protein,but no association expression of Epac1 and PKC protein,Epac1 and Cx43 protein is found in breast cancer tissues.It is further verified that the mechanism of Epacl regulating cancer progression is tissue specific.The second part is cell biology experiment.Objective:To explore the effect of binding of AKAP95 and Cx43 protein,AKAP95 and CyclinE2 protein on cell cycle progression.Methods:The BEAS-2B cell was selected as the research object.CCK-8 was used to detect the cell proliferation.Western Blot and Immunoprecipitation methods were used to detect the changes of AKAP95,Cx43 and CyclinE2 protein expression levels as well as the binding amount of AKAP95 and Cx43 protein,AKAP95 and CyclinE2 protein when platelet derived growth factor PDGF-BB is added to the cell,and further study the protein kinase which affected the binding amount.Conclusion:1.PDGF-BB can promote cell proliferation and the expression of AKAP95,Cx43,CyclinE2 protein in BEAS-2B cells.2.The combination of AKAP95 and Cx43 protein inhibits the proliferation of normal lung epithelial cells.The binding of AKAP95 and CyclinE2 proteins can promote the proliferation of normal lung epithelial cells,suggesting that the Cx43 and Cyclin E2 proteins may be competing to combine with AKAP95 proteins during the cell cycle process,and the process may be mediated by protein kinase Erkl/2.