| Objective1.To investigate the value of exosomal mi RNA-21,free mi RNA-21 and EGFR gene mutations and lung cancer-related tumor markers CEA,SCCA and CYFRA21-1 in NSCLC risk prediction.2.To analyse the correlation of exosomal mi RNA-21 expression with the pathological type,pathological stage,and TNM stage of NSCLC.Methods1.5m L EDTA anticoagulated Peripheral blood samples were collected from 30 cases of NSCLC patients(test group)and 30 healthy persons(control group).Extracted exosomes from all 60 serum samples.Transmission electron microscopy and Western blot were used to identify the exosomes.2.q RT-PCR was used to detect the expression of exosomal mi RNA-21 and free mi RNA-21 in 30 NSCLC serum samples and 30 healthy human serum samples.Liquid chip analysis system was used to detect the Contents of tumor markers CEA,SCCA,and CYFRA21-1 in 30 NSCLC serum samples and 30 healthy human serum samples.Digital PCR was used to detect the mutations of lung cancer driver gene EGFR in 30 NSCLC serum samples and 30 healthy human serum samples.3.SPSS22.0 statistical software was used to compare whether the above experimental results are statistically different in the experimental group and the control group,and to analyse the relationship between serum exosomeal mi RNA-21 and pathological type,pathological stage and TNM staging of NSCLC patients.The results of exosomal mi RNA-21,free mi RNA-21,and tumor markers were expressed as (?) ±s.The Rank sum test was used to compare the measurement data between the two groups,and the KW test was used to compare the measurement data between the two groups.The diagnostic efficiency was analyzed by using receiver operating characteristic(ROC)curves.Results1.The serum exosomes were successfully extracted and observed under transmission electron microscopy.The size of the exosomes obtained was approximately 40nm,which was consistent with the 30-100 nm reported in literatures.Western Blot results showed that the extracted exosomes were rich in surface markers CD61,CD81;2.The expression of exosomal mi RNA-21 in NSCLC group was 119.64 ± 98.35 and 39.10 ± 34.12 in healthy group.The difference was statistically significant(P<0.05).The ROC curve was used to calculate the diagnostic value of NSCLC risk,the area under the curve was 87.6%.While the expression of free mi RNA-21 in NSCLC group was 38.36 ± 24.29 and 29.74 ± 23.80 in healthy group.and there was no significant difference between two groups(P<0.05);3.The mutation rate of EGFR gene in NSCLC group was 43.3%.There was no mutation in the EGFR gene in healthy group.The use of EGFR gene mutations for NSCLC risk prediction has a specificity of 100% and a sensitivity of 43.3%;4.By plotting the ROC curve,the area under the curve for CEA,SCCA,and CYFRA21-1 was 52.1%,65.0%,and 60.9%,respectively.The diagnostic efficiency was low,but if the three indicators were jointly diagnosed,the area under the curve increased significantly,being 77.3%.But still lower than 86.7% of serum exosomal mi RNA-21;5.The expression levels of serum exosomal mi RNA-21 in NSCLC patients were 119.17 ± 61.36 in adenocarcinoma,154.91 ± 168.67 in squamous cell carcinoma,80.34 ± 36.20 in large cell carcinoma,there was no statistical difference(P>0.05);The expression levels in poorly differentiation was 146.86 ± 72.61,in highly differentiation was 112.19 ± 39.40,with statistically significant difference(P<0.05);the expression levels in different TNM stages were: Phase I 85.28 ± 68.30,Phase II 113.51 ± 57.78,Phase III 63.54 ± 23.59,Phase IV 12.94 ± 63.78,there was no statistical difference(P>0.05).Conclusion1.Exosomal mi RNA-21 has a higher value in NSCLC risk prediction;free mi RNA-21,EGFR gene mutations,and lung cancer-related tumor markers do not have predictive value for NSCLC risk;2.The expression of exosomal mi RNA-21 in patients with NSCLC is associated with pathological stage,but not with pathological type and TNM stage,Poorly differentiated cancer has higher expression of exosomal mi RNA-21. |
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