Fig. 3: Comparative analysis of plasma-derived miRNAs in LUSC patients and healthy individuals.

a Heatmap and boxplot indicate DE miRNA expression (n = 28). b Correlation plot of DE miRNA expression levels. Similar variables are placed adjacently using correlation-based variable ordering. Darker colours and larger circles indicate stronger correlations. Blue indicates a positive correlation, while red represents a negative correlation. c Heatmap and boxplot show DE miRNA expression (n = 28) in the validation cohort. Data is displayed as means ± SD; statistical evaluation using Student’s t-test and significant are miRNAs with p < 0.05. d Heatmap of DE miRNA expression in early disease stage (n = 13). e Heatmap of DE miRNA expression in late disease stage (n = 19). f Venn diagram depicting overlapped DE miRNA (n = 6) between early-stage vs. healthy (n = 13) and late-stage vs. healthy (n = 19), as well as stage-specific miRNAs in the early-stage (n = 7) and late-stage (n = 13). g ROC analysis reveals the best combination panel of DE miRNAs with the highest sensitivity (SE) and specificity (SP), as well as the best area under the curve (AUC) for the LUSC cohort. h Violin plot shows the probability density of the two compared sample groups (LUSC vs. healthy). i Pie chart shows the percentages of false predictions (false positives, FPs; false negatives, FNs) and true predictions (true positives, TPs; true negatives, TNs). j Table displays the best miRNA combination panel according to the highest AUC, SE, SP, and optimal cut-off in both LUSC and validation cohorts as determined by the CombiRoc analysis.