Role of Biomarkers Identified in Breast Cancer, As Possible Therapeutic and Diagnostic Agents with A Combination of Molecular and In-Silico Approaches

Abstract

Breast cancer is a heterogeneous disease for which strong biomarkers are needed, contributing to the improvement of diagnosis, prognostic classification, and therapy. This study investigated the diagnostic, prognostic, and therapeutic potential of miR-21, mesothelin (MSLN), and BCL2L11 (BIM) with an integrated molecular and in-silico strategy. The gene expression, protein expression, and circulating levels of biomarkers were assessed in a case–control study design to correlate with clinicopathological features and survival. Statistical analyses were group comparisons, receiver operating characteristic (ROC) analysis, multivariable logistic regression, association testing with molecular subtype, and survival analysis using Cox proportional hazards and Kaplan–Meier techniques. miR-21 (2.24 ± 0.65 log2FC) and MSLN mRNA (1.96 ± 0.61 log2FC) were significantly higher expressed in BC patients than healthy controls (p < 0.001 for both), whereas BCL2L11 expression was significantly decreased compared with healthy controls (−0.71 ± 0.52 vs 0.02 ± 0.34 log2FC, p < 0001). Serum MSLN levels were significantly higher in patients than in healthy individuals (29.6 ± 10.8 vs 7.6 ± 2.3 ng/mL; p < 0.001). ROC analysis showed excellent diagnostic value of serum MSLN (AUC 0.97) and miR-21 (AUC 0.96). In an age-adjusted model, including serum MSLN (OR = 1.42, 95% CI: 1.27–1.61, p <.001) and miR-21 (OR =3.68; 95% CI:2.11–6.42; p <.001), while BCL2L11 exerted a protective effect against breast cancer risk (OR =0.34; 95%CI:0.21 –0.55; p<.001). High serum MSLN was correlated with poorer PFS (HR = 1.05, 95% CI: 1.02–1.08, p = .002), whereas increased expression of BCL2L11 was associated with better PFS (HR = 0.71, 95% CI: 0.55–0.91, p =.007) and in silico docking revealed several ligands for MSLN, BCL2L11,, and PI3Kα with high binding affinities (up to −10.11 kcal/mol) and acceptable ADMET profiles. Together, these findings establish miR-21 and MSLN as promising diagnostic and prognostic biomarkers and identify BCL2L11 as a protective apoptotic marker with potential therapeutic implications in breast cancer.