Matrix metalloproteinase‑1 and microRNA‑486‑5p in urinary exosomes can be used to detect early lung cancer: A preliminary report

Abstract

The present study describes a novel molecular-genetic method suitable for lung cancer (LC) screening in the work-place and at community health centers. Using urinary-isolated exosomes from 35 patients with LC and 40 healthy volunteers, the expression ratio of MMP-1/CD63, and the relative expression levels of both microRNA (miRNA)-21 and miRNA-486-5p were measured. MMP-1/CD63 expression ratio was significantly higher in patients with LC than in the healthy controls {1.342 [95% confidence interval (CI): 0.890-1.974] vs. 0.600 (0.490-0.900); P<0.0001}. The relative expression of miRNA-486-5p in male healthy controls was significantly different from that in female healthy controls, whereas there was no significant difference in miRNA-21. Receiver operating characteristic curve (ROC) analysis of MMP-1/CD63 showed 92.5% sensitivity and 54.3% specificity, whereas miRNA-486-5p showed 85% sensitivity and 70.8% specificity for men, and 70.0% sensitivity and 72.7% specificity for women. The logistic regression model used to evaluate the association of LC with the combination of MMP-1/CD63 and miRNA-486-5p revealed that the area under the ROC curve was 0.954 (95% CI: 0.908-1.000), and the model had 89% sensitivity and 88% specificity after adjusting for age, sex and smoking status. These data suggested that the combined analysis of MMP-1/CD63 and miRNA-486-5p in urinary exosomes may be used to detect patients with early-stage LC in the work-place and at community health centers, although confirmational studies are warranted.

Keywords: MMP-1; lung cancer screening; miRNA-486-5p; urinary exosome.

Figure 1.

Comparison of the expression ratio of MMP-1/CD63 in 35 patients with LC and 40 healthy controls (20 males and 20 females). (A) Expression ratio of MMP1/CD63 in 35 patients with LC and 40 healthy controls (all). (B) Expression ratio of MMP1/CD63 in 24 male patients with LC and 20 male healthy controls. (C) Expression ratio of MMP1/CD63 in 11 female patients with LC and 20 female healthy controls. (D) Expression Ratio of MMP-1/CD63 in the patients with LC based on cancer stages (all). (E) Expression ratio of MMP1/CD63 in 24 male patients with LC based on cancer stages. (F) Expression ratio of MMP1/CD63 in 11 female patients with LC based on cancer stages. (G) Expression ratio of MMP1/CD63 in all patients with LC based on smoking habits (all). (H) Expression ratio of MMP1/CD63 in 24 male patients with LC based on smoking habits. (I) Expression ratio of MMP1/CD63 in 11 female patients with LC based on smoking habits. Median and 95% confidence intervals for the results were indicated by bars in the plots. LC, lung cancer; MMP-1, matrix metalloproteinase-1; ns, not significant.

Figure 2.

Comparison of relative expression of miRNA-21 in 35 patients with LC and 40 healthy controls (20 males and 20 females). (A) Relative expression of miRNA-21 in 35 patients with LC and 40 healthy controls (all). (B) Relative expression of miRNA-21 in 24 male patients with LC and 20 male healthy controls. (C) Relative expression of miRNA-21 in 11 female patients with LC and 20 female healthy controls. (D) Relative expression of miRNA-21 in the patients with LC based on cancer stages (all). (E) Relative expression of miRNA-21 in 24 male patients with LC based on cancer stages. (F) Relative expression of miRNA-21 in 11 female patients with LC based on cancer stages. (G) Relative expression of miRNA-21 in all patients with LC based on smoking habits (all). (H) Relative expression of miRNA-21 in 24 male patients with LC based on smoking habits. (I) Relative expression of miRNA-21 in 11 female patients with LC based on smoking habits. Median and 95% confidence intervals for the results were indicated by bars in the plots. LC, lung cancer; miRNA, microRNA; ns, not significant.

Figure 3.

Comparison of the Relative Expression of miRNA-486-5p in 35 patients with LC and 40 healthy controls (20 males and 20 females). (A) Relative expression of miRNA-486-5p in 35 patients with LC and 40 healthy controls (all). (B) Relative expression of miRNA-486-5p in 24 male patients with LC and 20 male healthy controls. (C) Relative expression of miRNA-486-5p in 11 female patients with LC and 20 female healthy controls. (D) Relative expression of miRNA-486-5p in 40 patients with LC based on cancer stages (all). (E) Relative expression of miRNA-486-5p in 24 male patients with LC based on cancer stages. (F) Relative expression of miRNA-486-5p in 11 female patients with LC based on cancer stages. (G) Relative expression of miRNA-486-5p in all patients with LC based on smoking habits (all). (H) Relative expression of miRNA-486-5p in 24 male patients with LC based on smoking habits. (I) Relative expression of miRNA-486-5p in 11 female patients with LC based on smoking habits. Median and 95% confidence intervals for the results were indicated by bars in the plots. LC, lung cancer; miRNA, microRNA; ns, not significant.

Figure 4.

ROC analysis. (A) ROC curves of the expression ratio of MMP-1/CD63 in urinary exosomes from 35 patients with LC and 40 healthy controls. (B) ROC curves of relative expression of miRNA-486-5p in urinary exosomes from 24 male patients with LC and 20 male healthy controls. (C) ROC curves of relative expression of miRNA-486-5p in urinary exosomes from 11 female patients with LC and 20 female healthy controls. mir-486; microRNA-486-5p. (D) ROC analysis for the usefulness of both biomarkers in LC screening via logistic regression analysis. AUC was 0.954 (95% CI: 0.908–1.000). AUC, area under the ROC curve; miRNA, microRNA; MMP-1, matrix metalloproteinase-1; LC, lung cancer; ROC, receiver operating characteristic.