The Role Of Macrophage Activated By Lung Cancer Derived Exosomes In Lung Cancer Progression

Objective: Lung cancer is one of the most common malignancies in the world.The incidence rate is high,the incidence is concealed,early diagnosis is insufficient and prone to metastasis,80% of patients are in the middle and late stage at the first diagnosis.Although the advancement of diagnostic methods,the improvement of surgical techniques,the application of radiotherapy and chemotherapy,and the application of targeted drugs have improved the survival rate of patients to a certain extent,the effect is not obvious,so it is necessary to conduct in-depth research on the biological behavior of lung cancer.Tumor cells can regulate the tumor microenvironment by releasing exosomes into the surrounding environment,with the aim of forming a microenvironment that is beneficial to tumor growth,metastasis and invasion.Macrophages,as the most abundant inflammatory cells in the tumor microenvironment,can be regulated by tumor cells and can exhibit diverse effects.Therefore,the purpose of this paper is to investigate the effect of lung cancer derived exosomes on macrophage polarization and the biological effects of macrophages on lung cancer after exosome treatment,providing a certain basic research further study for the biological behavior of tumors.Methods: The culture supernatant of lung cancer cells was collected,and the culture supernatant was concentrated by ultrafiltration.The exosomes were extracted by reagent method,and the morphology of exosomes was identified by transmission electron microscopy and nanometer particle size.The surface markers of exosomes were identified by Western blot.When THP-1 cells were cultured in good growth state,they were treated with 100 ng/ml PMA for 2 days,the cells were adherent,the macrophage marker CD68 was detected by real-time PCR,then the macrophage was detected Phagocytosis function;macrophages were co-cultured with exosomes(200?g/ml)for 24 h,then qPCR was used to detect macrophage phenotype molecules INOS and CD163,and chemoluminescence was used to detect IL-10,IL-6 and IL-8 expression;macrophage after exosome treatment were co-cultured with lung cancer cells by transwell chamber co-culture system,lung cancer after co-cultured were used cell invasion assay.MMP9,MMP2 and E-cadherin were detected by real-time PCR.Results: The small vesicles were diameter of about 130 nm by transmission electron microscopy and nano-particle size analysis.The high expression of CD9 and CD63 in the extracted exosomes was detected by Westen blot,indicating that we successfully isolated exosomes from the tumor culture supernatant;macrophages induced by PMA were highly expressed CD68 by real-time PCR,macrophage phagocytosis experiments showed that exosomes can be phagocytosed by macrophages,indicating that macrophages induced by PMA can be used for subsequent experiment.The macrophages after exosome treatment were detected by real-time PCR,the expression of INOS was decreased,the expression of CD163 was increased,and the macrophages after exosome treatment were detected by chemiluminescence,the secreted IL-10,IL-6 and IL-8 were all increased.cell invasion assays found that lung cancer cells co-cultured with macrophages after exosome treatment highly expressed MMP9,MMP2 and E-cadherin.Conclusion: 1.Lung cancer derived exosomes can induce macrophage polarization,not a single phenotype,but a mixed type of both M1 and M2 types,and more M2 types.2.Lung cancer can further enhance its invasion and metastasis ability by regulating macrophages in the microenvironment through the regulation of macrophages in the microenvironment.