BTK Regulates TAM Activation Through The Raf/ERK Signal Axis To Promote Lung Cancer Cell Migration And Invasion

Objective:The present study aims to investigate the impact of macrophage-expressed BTK on lung cancer cell migration and invasion and to elucidate the specific molecular mechanism underlying the promotion of lung cancer cell migration and invasion by BTK.Our findings aim to provide a novel idea and theoretical basis for the treatment of lung cancer targeted against BTK.Methods:The expression of BTK in various cancers was analyzed using a network database.The survival and prognosis of lung adenocarcinoma(LUAD)and lung squamous cell carcinoma(LUSC)with inconsistent expression with other solid tumors were analyzed.The cellular localization of BTK was observed by immunofluorescence staining of lung tissue.We successfully constructed sh BTK-THP-1 cell line by lentivirus transfection.Western blot and RT-PCR were used to detect the differences in BTK protein and m RNA levels,and the corresponding supernatant was collected as conditioned medium.The effects of BTK gene on the migration and invasion ability of lung cancer cells were evaluated using scratch test and Transwell test.The expression of chemokines(CCL2,CCL5)was detected by RT-q PCR.The effect of Raf/ERK/NF-κB signaling pathway on migration and invasion of lung cancer cells was determined by detecting the expression of p-Raf,p-ERK,p-NF-κB,and ERK,NF-κB inhibitor validation.Animal experiments were designed to detect the effect of BTK inhibitor(Ibrutinib)in a C57BL/6 mice in situ lung cancer animal model,and the effect of Ibrutinib on tumor was evaluated by tumor size and lung respiratory function.Results:Bioinformatics analysis revealed that the expression level of BTK gene was relatively low in LUAD patients,and it was positively correlated with the prognosis of LUAD patients.Tissue immunofluorescence staining showed that BTK was predominantly localized on macrophages in the tumor nest and adjacent to the tumor,and its expression increased with an increase in the number of macrophages around the tumor.The expression of BTK in macrophages significantly enhanced the migration and invasion ability of lung cancer cells and upregulated the expression of CCL2 and CCL5.In the subsequent mechanism study,BTK silencing was found to significantly inhibit the Raf/ERK signal axis in the MAPK signal pathway,while downstream p-NF-κB transcription factor and ERK inhibitor also inhibited the migration and invasion of lung cancer cells.In vivo,administration of Ibrutinib,an inhibitor of BTK,significantly suppressed the growth of lung cancer and improved lung function in mice.Conclusion:Our study revealed that BTK was highly expressed in tumor-associated macrophages of lung cancer and played a critical role in regulating the expression of CCL2 and CCL5 through the Raf/ERK/NF-κB pathway,ultimately promoting lung cancer cell migration and invasion.Importantly,we also demonstrated that Ibrutinib efficiently inhibited lung cancer growth and improved lung function in mice,indicating that BTK may be a promising target for the treatment of lung cancer.Figure [six] Table [fifteen] Reference [forty-seven]...