The Nf-?B Signaling Pathway Activated By IL-1? Regulates The Sorting Of Lung Squamous Cell Carcinoma Exosome MiR-223-3p

Background Lung squamous cell carcinoma is a common pathological type of lung cancer,and its pathogenesis and treatment progress are obviously lagging behind.Chronic inflammation can lead to canceration,and the role of inflammation in the occurrence and development of lung cancer has been gradually recognized.More and more studies have shown that miRNAs can be an important link between inflammation and tumor,and participate in the occurrence and development of tumors.Extracellular vesicles are important mediators of intercellular communication in tumor microenvironment.Exosomes as an important class of extracellular vesicles,especially tumor derived exosomes have received more and more attention.Exosomes contain a large number of miRNAs,and the loading and secretion of exosomes miRNAs are affected by microenvironmental factors.Inflammation is a basic feature of tumor microenvironment,but there are few reports on the effect of inflammatory factors on tumor exosomes miRNAs.As an important inflammatory factor,the role of IL-1? in the selective loading and secretion of lung squamous cell carcinoma exosomes miRNAs is an important scientific question.Objective In this study,we aimed to explore the mechanism by which the inflammatory factor IL-1? regulates the expression of miR-223-3p in exosomes of lung squamous cell carcinoma,providing theoretical basis and potential targets for the anti-inflammatory treatment of lung squamous cell carcinoma.Methods(1)Plasma IL-1? levels were determined by flow cytometry in 20 patients with lung squamous cell carcinoma and 20 healthy controls.(2)q RT-PCR was used to detect the effect of IL-1? on the expression of miR-223-3p in and outside SK-MES-1 cells of lung squamous cell carcinoma.(3)Exosomes of lung squamous cell carcinoma SK-MES-1 cells(treated and untreated by IL-1?)were extracted,WB was used to detect exosome-related protein expression for preliminary identification of exosomes,and the expression levels of miR-223-3p in the exosomes of cells which treated or untreated by IL-1? were detected by q RT-PCR.(4)Expression of related proteins(p-I?B? I?B? and p-p65)and YBX1 in the NF-?B signaling pathway of SK-MES-1 cells treated with IL-1? was detected by Western blot and immunofluorescence.Protein expression of p-p65 and YBX1 in SK-MES-1 cells pretreated with NF-?B kinase inhibitor(I?B? kinase inhibitor,BAY 11-7082)was also detected.(5)q RT-PCR was used to detect the effect of IL-1? on the expression of miR-223-3p in and out of lung squamous cell carcinoma cells after down-regulation of YBX1.(6)Confocal microscopy was used to observe the distribution of p27kip1 in lung squamous cell carcinoma SK-MES-1 cells,and Western blot was used to detect the effect of IL-1? on the expression of p27kip1 in lung squamous cell carcinoma SK-MES-1 cells.Results(1)The median plasma IL-1? content in lung squamous cell carcinoma patients was 11.96(7.55,29.20)ng/L,higher than that in healthy controls 2.52(1.64,3.48)ng/L(P<0.001).(2)After IL-1? treatment of lung squamous cell carcinoma SK-MES-1 cells,the expression of Mir-233-3p in the cells was decreased,but the expression of Mir-223-3p in the superfluid was increased.(3)miR-223-3p expression was increased in culture exosomes of SK-MES-1 cell treated by IL-1?.(4)These were Increased expression of p-I?B?,decreased expression of I?B?,increased expression of p-p65, increased expression of nuclear p65,and increased expression of YBX-1 in the NF-?B signaling pathway after IL-1? treatment.The above phenomenon were inhibited after pretreatment with NF-?B signaling pathway inhibitors.(5)After the down-regulation of YBX1 protein expression,the effect of IL-1? on the expression of miR-223-3p inside and outside lung squamous cell carcinoma SK-MES-1 cells was inhibited.Conclusions IL-1? promotes the expression of YBX1 protein in lung squamous cell carcinoma cells by activating the NF-?B signaling pathway,which secretes the formation of intracellular miR-223-3p exosomes into the extracellular environment,resulting in decreased expression of intracellular miR-223-3p.The results of this study are of great significance to further study the mechanism of inflammation promoting tumor progression.