The Function And Mechanism Of Plasma Exosomes To Promote Malignant Progression Of Lung Adenocarcinoma In Patients With Obstructive Sleep Apnea Syndrome

Objectives: The aim of this study is to detect the expression of mi RNA-33b-3p in plasma exosomes of patients with obstructive sleep apnea hypopnea.To explore the role of exosome mi RNA-33b-3p/AXL in affecting the proliferation and migration of lung adenocarcinoma cells.Method:(1)Separate exosomes from peripheral blood of OSA patients and healthy subjects,Further characterization was confirmed by Nanosight nanoparticle tracking analysis technology and transmission electron microscopy.At the same time,the expression of specific exosomal proteins CD63,CD9 and Hsp70 was detected by Western blot.Further,q PCR was used to detect the difference in expression of exosome mi RNA-33b-3p between the two groups.(2)The exosomes are co-incubated with PKH26-labeled exosomes and lung adenocarcinoma A549 cell lines to identify exosomes that could be taken up by A549 cells.Meanwhile,A549 cells are transfected with fluorescently labeled mi RNA-33b-3p,and the exosomes containing the specific mi RNA are incubated with common A549 cells.(3)The A549 cells are incubated with the exosome of the OSA patient group and the healthy human group,and the corresponding mi RNA-33b-3p levels and cell proliferation and migration ability were compared.(4)To verify the role of mi RNA-33b-3p in affecting the proliferation and migration of A549 cells,mi RNA-33b-3p mimics and their mi RNA-33b-3p mimics are transfected into A549 cells,respectively,and compare their proliferation and migration ability.(5)In order to verify that AXL is a mi RNA-33b-3p-acting protein and play a role in the malignant increase of A549 cells induced by mi RNA-33b-3p,luciferase reporter gene assay and cell experiments were performed.Transfection of mi RNA-33b-3p inhibitor in A549 cells attenuated its regulatory function,interfer with the expression of AXL protein,and explore the role of target cell proliferation and migration in different situations to further validate the role of AXL in the pathogenesis of mi RNA-33b-3p affecting the malignant progression of A549 cells.Result:(1)Exosomes are isolated from peripheral blood plasma of OSA patients and healthy subjects,and the diameter was concentrated in nm.CD63,CD9,and Hsp70 expression were observed in WB assay.q PCR detection showed that mi RNA-33b-3p was less expressed in OSA patients than in healthy subjects.(2)Exosomal mi RNAs can be uptaked by A549 cells along with exosomes.(3)Peripheral blood exosomes in OSA patients can promote the proliferative and migration ability of the target cells together with healthy subjects.(4)mi RNA-33b-3p attenuates the proliferation and migration of A549 cells.(5)AXL protein is a direct acting protein of mi RNA-33b-3p to attenuate the malignant progression of A549 cells.Conclusion: Compared with healthy individuals,the expression of mi RNA-33b-3p in peripheral blood exosomes of OSA patients is reduced.The exosome-mi RNA-33b-3p/AXL pathway is one of the mechanisms for OSA patients to promote the proliferation and migration of lung adenocarcinoma cells.