| Objecctive: Lung cancer is leading cause of cancer-related death worldwide.Due to lack of early clinical manifestations and specific early screening methods,most patients are diagnosed as advanced.lung adenocarcinoma(LUAD)is an important component of lung cancer.In view of the deficiency of traditional tumor molecular markers,this study aimed to find out the serum exosome mi RNAs with potential diagnostic value for lung adenocarcinoma by the serum exosome mi RNAs microarray and Real-time fluorescence quantitative PCR in patients with LUAD,and explore its preliminary functional in LUAD.Mothods: Molecular markers and morphology of exosomes were performed by Western blot and transmission electron microscopy(TEM);Microarray technology was used to analyze differentially expressed mi RNAs in serum exosomes of LUAD patients;Five mi RNAs with high expression and Five mi RNAs with low expression were selected for preliminary verification by real-time fluorescent quantitative PCR;Expression levels of mi RNAs with highest and lowest were selected for further validation,and explored the correlation between mi RNAs expression levels and clinical parameters;Receiver operating characteristic curve(ROC)was used to assess the diagnostic efficacy of mi RNAs;Finally,A549 cell lines with low expression of mi R-7977 were constructed by mi RNA inhibitor transfection.The effects of mi R-7977 on proliferation,invasion and apoptosis of A549 were detected by flow cytometry,CCK8,and invasion assay.Results: Results of Western blot showed that the extracted substances expressed exosome molecular markers CD9 and CD63,their size and morphology were consistent with exosomes under TEM.Microarray analysis showed that 31 mi RNAs were up-regulated and 29 mi RNAs were down-regulated in serum exosomes of lung cancer patients.Subsequently,five up-regulated(mi R-451 a,mi R-7977,mi R-1260 a,mi R-7641,mi R-197-3p)and five down-regulated mi RNAs(mi R-133 b,mi R-1185-1-3p,mi R-4284,mi R-4463,mi R-98-3p)were preliminarily verified by real-time fluorescent quantitative PCR.The results showed that mi R-451 a,mi R-7977,mi R-7641 were up-regulated and mi R-98-3p,mi R-133 b were down-regulated in LUAD patients.Mi R-7977 and mi R-98-3p were selected to further expand the sample validation,results showed that compared with the control group mi R-7977 was higher expressed(p < 0.05)and mi R-98-3p was lower expressed(p < 0.05).The expression level of mi R-7977 was significantly associated with N stage and TNM stage(p < 0.05).The area under the ROC curve(AUC)of mi R-7977 and mi R-98-3p were 0.787(95% CI: 0.705-0.855)and 0.719(95% CI: 0.632-0.796),respectively.When combined,the area under the curve was 0.816(95% CI: 0.737-0.880),and the combined diagnostic efficiency was significantly better than the individual indicators.Compared to normal lung epithelial cells(Beas-2b)mi R-7977 was significantly higher expressed in LUAD cells(A549).A549 with low expression of mi R-7977 was constructed by mi R-7977 inhibitor transfection,the results showed that low expression of mi R-7977 significantly enhanced the proliferation and invasion and inhibited apoptosis in A549 cells.Conclusion: The expression of mi R-7977 was significantly high-expression in serum-derived exosomes of LUAD,and the expression of mi R-7977 was significantly associated with N stage and TNM stage.Similarly,the expression level of mi R-7977 in A549 was higher than that of Beas-2b.Decreasing the expression of mi R-7977 in A549 cells could promote the proliferation of A549 cells,inhibit its apoptosis and enhance its invasion ability.The serum-derived exosome mi R-7977 has potential as a molecular marker for the diagnosis of lung adenocarcinoma,and mi R-7977 may be a target for the treatment of lung adenocarcinoma. |
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