Inhibition Of ANXA2 In Liver Cells Attenuates The Lung Cancer Liver Metastasis

ObjectiveCurrent clinical high incidence of lung cancer and liver metastases refractory makes targeted drug therapy,research transfer mechanism in this direction is particularly important,and liver metastases of lung cancer specimens in clinical studies is difficult to become a huge accessibility of the research process obstacle.Application of conventional transwell platform to build a lung adenocarcinoma cells A549,fibroblasts WI38,monocytemacro-phage THP-1 cells and human liver cells L-02 co-culture system,namely lung cancer liver metastasis model to solve the problem which is difficult to obtain clinical specimens,In the initial process of lung cancer liver metastasis,the key molecule ANXA2 in hepatocytes was screened by proteomics techniques and its role in lung cancer liver metastasis process is studied by wound healing assay and chemotaxis assay.This study elaborated a novel mechanism concerned about lung cancer liver metastasis,with a view to the application of proteomics and traditional platforms in metastasis,targeted therapy and tumor microenvironment of tumor biology.Method1.A transwell platform is built to duplicate the lung cancer liver metastases process,and its reliability is validated by detecting protein E-cad,Snaillin A549 cells and AFP in liver cells L-02 by Immunofluorescence and westernblot methods.We set up this model in order to obtain a large number of protein samples for subsequent analysis.2.Protein samples were analyzed by the usage of proteo-mics mass spectrometry(iTRAQ technology)?nd the differently expressed proteins in liver cells were screened out before and after lung cancer colonization.The candidate list of differentially express-ed protein was combined with the mRNA expression data,protein-protein interaction data,KEGG and GO annotation data,the importance of differentially expressed protein were analyzed from multiple perspectives at different levels,and ultimately we selected protein ANXA2 in liver cells for next step verification.3.Downregulation of ANXA2 in liver cells with shRNA and the metastasis capacity of lung cancer cells was evaluated by a wound healing assay and chemotaxis assay.4.Lung metastases attached an example of typical case analysis,diagnosis and treatment from the entire process and the current diagnosis,treatment discussions with a view to better illustrate the clinical application of the subject,deepen the current situation at the diagnosis and treatment of lung cancer and liver metastases still need research to understand.ResultIn this study,a lung cancer liver metastasis model was established to duplicate the initial process of the colonization of the lung cancer cells in liver by detecting cancer-associated proteins.ANXA2,a candidate protein was screened and the functional evaluation was performed with proteomics mass spectrometry.It was confirmed that after knockdown ANXA2 protein content in liver cells,the athletic ability of lung cancer cells and chemotactic ability of lung cancer cells to liver cells were decreased,which means the knockdown of ANXA2 in liver cells inhibit metastasis of lung cancer,which provides a new method for experimental and clinical study of lung metastases of other tumor target organs as well as the prevention and treatment of tumor invasion and metastasis in vitro.