Therapeutic targeting of ROCK reverses EMT in lung cancer cells by impeding TAZ in Hippo signalling pathway.

RRM2 promotes lung adenocarcinoma progression and is associated with ferroptosis-inducer sensitivity through the NRF2/GPX4 signaling axis.

BACKGROUND Lung adenocarcinoma (LUAD) is a leading cause of cancer-related mortality, characterized by aggressive progression and therapy resistance. Ferroptosis, an iron-dependent form of regulated cell death, has emerged as a promising therapeutic avenue. However, the role of Ribonucleotide Reductase M2 (RRM2) in ferroptosis regulation and its relevance to LUAD progression remain incompletely understood. METHODS We integrated bulk transcriptomic, proteomic, WGCNA, and single-cell datasets to evaluate the clinical and biological relevance of RRM2 in LUAD. Functional validation was performed using RRM2 knockdown, ferroptosis-inducer sensitivity assays, ferroptosis-related biochemical assays, NRF2/GPX4 pathway analysis, rescue experiments, and xenograft models. RESULTS RRM2 was significantly upregulated in LUAD tissues and was associated with poor overall survival. Single-cell analysis localized high RRM2 expression to a proliferative tumor cell subpopulation enriched in cell cycle- and immune-related pathways. Functionally, RRM2 knockdown suppressed LUAD cell proliferation and tumor growth and was accompanied by increased ROS, lipid ROS, Fe²⁺, and MDA levels and decreased GSH levels. RRM2 depletion also increased ferroptosis-inducer sensitivity, with enhanced erastin and RSL3 sensitivity in A549 cells and clear RSL3 sensitization in PC9 cells. In parallel, RRM2 silencing was associated with reduced NRF2 and GPX4 expression, decreased NRF2 nuclear-to-cytosolic signal intensity, and increased ACSL4 expression. NRF2 overexpression partially restored GPX4 immunofluorescence intensity in RRM2-knockdown cells. Moreover, NRF2 overexpression or Ferr-1 treatment partially reversed the growth-suppressive effects induced by RRM2 deficiency in vitro and in vivo. CONCLUSION RRM2 is associated with LUAD progression, ferroptosis-inducer sensitivity, and ferroptosis-related phenotypes, potentially through modulation of the NRF2/GPX4 axis. These findings support RRM2 as a candidate prognostic biomarker and a potential therapeutic target in LUAD, while the precise molecular relationship between RRM2 and the NRF2/GPX4 axis warrants further investigation.

MICAL2 as a key effector of β-catenin signaling driving melanoma progression and therapeutic resistance.

Potential multi-target inhibition of the PI3K-Akt pathway by Fraxetin suppresses colorectal cancer and restores 5-FU sensitivity in 5-FU-resistant CRC cells: An integrated study combining network pharmacology, molecular simulation, and in vitro validation.

Colorectal cancer (CRC) is a malignancy with high global incidence and significant therapeutic challenges, where chemotherapy resistance is a key factor leading to treatment failure. Aberrant activation of the PI3K-Akt signaling pathway plays a central role in CRC progression and drug resistance. Fraxetin, a major active coumarin component of the traditional Chinese medicine Cortex Fraxini, possesses various biological activities, but its anti-CRC effects and underlying mechanisms remain unclear. This study aimed to systematically elucidate the mechanism of action of Fraxetin against CRC, particularly its regulation of the PI3K-Akt pathway and potential to reverse drug resistance, by integrating network pharmacology, computational simulation, and in vitro experiments. Network pharmacology screening identified 108 common targets of Fraxetin and CRC. Protein-protein interaction (PPI) analysis pinpointed 10 core targets, including TNF, AKT1, and EGFR. KEGG enrichment analysis suggested the PI3K-Akt pathway as one of the primary pathways involved. Molecular docking and dynamics simulations confirmed that Fraxetin could stably bind to core targets such as EGFR and ERBB2. In vitro experiments demonstrated that Fraxetin dose-dependently inhibited the proliferation of HCT116 and HT-29 cells, induced reactive oxygen species (ROS) generation, and significantly downregulated the phosphorylation levels of key PI3K-Akt pathway proteins, p-PI3K and p-Akt. Furthermore, Fraxetin combined with 5-fluorouracil (5-FU) or irinotecan exhibited synergistic antiproliferative effects and significantly restored the sensitivity of 5-FU-resistant cells to chemotherapeutic agents. This study systematically demonstrate that Fraxetin exerts multi-faceted effects against colorectal cancer, including anti-cancer activity, synergy with chemotherapy, and restoration of 5-FU sensitivity in 5-FU-resistant CRC cells in vitro, through multi-target inhibition of the PI3K-Akt signaling pathway.

Pharmacological inhibition of BUB1 suppresses UCEC progression by modulating the PI3K-AKT pathway and reversing anoikis resistance and immune evasion: a multi-omics and experimental study

Uterine corpus endometrial carcinoma (UCEC) is a prevalent gynecological malignancy with poor prognosis and limited therapeutic strategies. The mitotic checkpoint kinase BUB1 has emerged as a potential regulator of tumor progression, but its clinical and functional roles in UCEC remain unclear. Transcriptomic datasets from TCGA and GEO were analyzed to evaluate BUB1 expression, survival impact, pathway enrichment, methylation, and immune infiltration. Protein–protein interactions and drug sensitivity analyses were performed using bioinformatics pipelines. Functional validation was conducted in HEC-1-B cells treated with the BUB1 inhibitor 2OH-BNPP1 using MTT, wound healing, and qPCR assays. BUB1 was significantly upregulated in UCEC tumors compared with normal endometrium, associated with advanced disease stage and reduced overall survival, confirming its role as an unfavorable prognostic biomarker. Enrichment analyses linked BUB1 to cell cycle regulation, PI3K-AKT signaling, and anoikis resistance. BUB1 expression positively correlated with infiltration of macrophages, neutrophils, and CD4⁺ T cells. Pharmacological inhibition with 2OH-BNPP1 reduced cell viability, impaired migration, and suppressed BUB1 expression in a dose-dependent manner, supporting its oncogenic role in endometrial carcinoma. This study identifies BUB1 as a clinically relevant prognostic biomarker and therapeutic target in UCEC. Targeting BUB1 interrupts tumor-promoting processes including proliferation, survival, and migration, underscoring its translational potential for future therapeutic development. BUB1 is significantly overexpressed in UCEC and correlates with poor survival and advanced tumor stage. BUB1 promotes anoikis resistance and may drive metastatic potential in UCEC cells. Hypomethylation, SNV and CNVs contribute to the overexpression of BUB1 in UCEC tumors. BUB1 expression is positively associated with immunosuppressive cell infiltration. 2OH BNPP1 inhibits BUB1, thereby reducing the viability and migration of UCEC cells in vitro. BUB1 is significantly overexpressed in UCEC and correlates with poor survival and advanced tumor stage. BUB1 promotes anoikis resistance and may drive metastatic potential in UCEC cells. Hypomethylation, SNV and CNVs contribute to the overexpression of BUB1 in UCEC tumors. BUB1 expression is positively associated with immunosuppressive cell infiltration. 2OH BNPP1 inhibits BUB1, thereby reducing the viability and migration of UCEC cells in vitro.

Epigenetic Remodeling Through GSK343-induced EZH2 Inhibition Alters SMYD2/SMYD3 Expression and Promotes Antitumor Effects.

BACKGROUND Breast cancer is a heterogeneous disease in which epigenetic dysregulation plays a critical role in tumor progression, therapeutic resistance, and cellular plasticity. Among epigenetic regulators, EZH2, the catalytic subunit of the polycomb repressor complex 2 (PRC2), has emerged as a key oncogenic driver through its role in H3K27 trimethylation (H3K27me3)-mediated transcriptional repression. This study aimed to evaluate the anti-tumor effects of the EZH2 inhibitor GSK343 and to investigate its impact on the expression of additional epigenetic regulators, SMYD2 and SMYD3, in breast cancer models. MATERIAL AND METHODS Human breast cancer cell lines MDA-MB-231 (triple-negative) and MCF-7 (luminal) were treated with increasing concentrations of GSK343 (1-60 µM) for 24, 48, and 72 hours. Cell viability was assessed by MTT and Trypan Blue assays, while apoptosis was evaluated through caspase-3/7 activity. Gene expression levels of EZH2, SMYD2, and SMYD3 were quantified by RT-qPCR, and H3K27me3 levels were analyzed as a pharmacodynamic marker of EZH2 inhibition. RESULTS GSK343 induced a significant dose- and time-dependent reduction in cell viability and a corresponding increase in apoptotic activity, with more pronounced effects in MDA-MB-231 cells. Treatment also resulted in consistent downregulation of EZH2, SMYD2, and SMYD3, alongside a marked decrease in H3K27me3 levels, confirming effective epigenetic modulation. These findings demonstrate that EZH2 inhibition promotes coordinated epigenetic remodeling and disrupts key oncogenic pathways in breast cancer cells. The greater sensitivity observed in triple-negative cells highlights subtype-specific epigenetic dependencies and supports EZH2 and SMYD family members as promising therapeutic targets. CONCLUSION EZH2 inhibition reveals subtype-specific vulnerabilities and supports targeting epigenetic regulators as a promising therapeutic strategy in breast cancer. Additionally, these results suggest potential combinatorial strategies integrating EZH2 inhibition with other targeted or epigenetic therapies to enhance treatment efficacy, overcome resistance mechanisms, and improve clinical outcomes, particularly in aggressive subtypes such as triple-negative breast cancer. Further studies are warranted to validate findings.

Targeting TAK1 to overcome cisplatin resistance in lung adenocarcinoma by rewiring the NF-κB and p53 signaling.

Acquired cisplatin resistance limits the efficacy of chemotherapy in advanced non-small cell lung cancer (NSCLC). Identifying key resistance regulators is clinically important. Although TAK1 (transforming growth factor-beta-activated kinase 1) has been implicated in drug resistance, its role in cisplatin-resistant lung adenocarcinoma remains unclear. Bioinformatic analysis of Gene Expression Omnibus (GEO) data identified TAK1 as a resistance-related gene. TAK1 expression was elevated in resistant clinical samples and cell lines (A549R, PC9R), positively correlating with the upregulation of drug-efflux proteins P-glycoprotein (P-gp) and ATP-binding cassette subfamily G member 2 (ABCG2). The TAK1 inhibitor (5Z)-7-oxozeaenol (5Z7) synergized with cisplatin to suppress proliferation and epithelial-mesenchymal transition, induce apoptosis, cause G0/G1 arrest, and promote senescence in resistant cells. Genetic TAK1 inhibition produced similar trends. Mechanistically, 5Z7 inhibited the TAK1-IκB kinase (IKK) axis, blocking inhibitor of κBα (IκBα) phosphorylation and nuclear factor kappa-B (NF-κB) p65 nuclear translocation, while upregulating p53 and its downstream effectors p21/p16-effects that were reversible by an NF-κB activator or p53 inhibitor. In vivo, 5Z7 plus cisplatin suppressed resistant tumor growth without notable organ toxicity. This study establishes TAK1 as a key hub in cisplatin resistance and shows that 5Z7 reverses resistance by targeting TAK1 to coordinately regulate NF-κB/p53 signaling, providing a foundation for potential clinical translation.