Matairesinol regulates TGF-β-induced EMT and TGF-β associated crosstalk with Wnt Signaling in PC3 prostate cancer cells: An omics-guided therapeutic insight.

Metastatic, castration-resistant prostate cancer (mPC) is driven by epithelial-to-mesenchymal transition (EMT), acquisition of stem-like traits, and resistance to apoptosis, processes orchestrated by TGF-β and reinforced by Wnt/β-catenin signallingsignaling. Effective agents that can simultaneously intercept these pathways remain scarce. Here, we investigated the dietary lignan Matairesinol (MAT) as a multitarget EMT antagonist in PC3 cells. RNA-seq followed by rigorous quality control (Phred >30; > 90 % unique mapping) identified 152 MAT-responsive transcripts (74 up-, 78 down-regulated). GeneCodis and Hallmark enrichment revealed MAT-mediated suppression of hypoxia response, NF-κB/TNF signaling, focal adhesion, and ECM-receptor interaction, alongside activation of p53 and senescence programs. Overlap with 24 cancer Hallmark gene sets yielded 56 prostate- cancer relevant DEGs, highlighting down-regulation of EMT-supportive pathways and up-regulation of tumor-suppressive networks. Functional validation in a TGF-β-induced EMT model showed that MAT (50, 100, and 150 μM) partially restored epithelial morphology, curtailed clonogenicity, and inhibited migration. JC-1 and ROS assays demonstrated reversal of EMT-associated apoptosis resistance via mitochondrial depolarization and oxidative-stress elevation. MAT also reduced mitochondrial biomass and dismantled actin stress fibers. Analysis by qRT-PCR and flow cytometry confirmed re-expression of E-cadherin, suppression of Vimentin/N-cadherin, and down-regulation of Snail, Twist, and Zeb1. Importantly, MAT lowered β-catenin while restoring GSK3β, inhibited downstream targets MYC and CCND1, shrank tumorspheres, and reduced CD44 expression. Collectively, MAT counters TGF-β/Wnt-driven EMT, stemness, and survival signaling at both transcriptomic and phenotypic levels. These findings nominate MAT as a promising scaffold for anti-metastatic therapy in advanced prostate cancer and warrant further in vivo evaluation.