The role of STRA6 induced by ONECUT2 drives malignant progression and radioresistance in rectal cancer through activating the Wnt/β-Catenin signaling pathway

GATA6-mediated transcriptional activation and METTL3/IGF2BP2-dependent post-transcriptional stabilization upregulate CDC20 to promote the malignant progression of colorectal cancer and immune escape

ZNF695 Promotes Colorectal Cancer Progression Through Transcriptional Activation of CBX8 and Subsequent Wnt/β-Catenin Signaling Activation.

In this investigation, we examined the functional mechanism of the transcription factor zinc finger protein 695 (ZNF695) and its target gene chromobox protein homolog 8 (CBX8) in colorectal cancer (CRC) migration and invasion. HCT-116 and LOVO cell lines were used to establish cell models with knocked-down ZNF695 and knocked-down or over-expressed CBX8. To comprehensively evaluate the functional contributions of ZNF695 and CBX8 to cellular phenotypes, we employed CCK-8, wound-healing, and Transwell assays to evaluate cell proliferation, migration, and invasion, respectively. To assess the impact of ZNF695 on tumor progression, we generated a xenograft model utilizing nude mice. A FLAG-ZNF695 expression plasmid was constructed, and ChIP-seq experiments were performed. By integrating mRNA sequencing data following ZNF695 knockdown with highly expressed genes in CRC from the TCGA database, CBX8 was identified as a putative downstream target of ZNF695. We employed a dual-luciferase reporter assay to validate the specific binding affinity of ZNF695 toward the CBX8 promoter region. To elucidate the specific biological cascades modulated by ZNF695 and CBX8, we conducted a comprehensive pathway enrichment analysis. Rescue experiments were conducted to determine whether the ZNF695/CBX8 regulatory axis upregulates the expression of the Wnt signaling pathway downstream targets, AXIN2 and CCND1. Both in vitro assays and in vivo models confirmed that silencing ZNF695 dramatically suppresses CRC cell proliferation, migration, and invasion, while concurrently impeding tumor progression. ChIP-seq coupled with dual-luciferase reporter assays substantiated the direct binding of ZNF695 to the CBX8 promoter. Furthermore, CBX8 depletion significantly attenuated the migratory and invasive phenotypes of CRC cells. Restoring CBX8 expression effectively rescued the migratory and invasive deficits in CRC cells induced by ZNF695 silencing. Re-expression of CBX8 in ZNF695-silenced cells restored Wnt/β-catenin signaling activity, accompanied by increased expression of AXIN2 and CCND1. ZNF695 promotes CRC progression by transcriptionally activating CBX8 and subsequently enhancing Wnt/β-catenin signaling, thereby promoting tumor cell proliferation, migration, and invasion.

HNRNPA0 Acts as a Master Upstream Regulator of NF-κB to Drive Oncogenic Phenotypes in Oral Squamous Cell Carcinoma

Background Oral squamous cell carcinoma (OSCC) remains a lethal malignancy with a 5-year survival rate below 50%, necessitating the identification of novel therapeutic targets. While the RNA-binding protein HNRNPA0 is implicated in various cancers, its precise role and mechanism in OSCC pathogenesis are undefined. Methods HNRNPA0 expression was assessed using TCGA data, clinical OSCC specimens (n = 10) and cell lines. Stable knockdown models were established in CAL-27 and SCC-15 cells via shRNA. Functional impacts on proliferation, metastasis, apoptosis and cell cycle were evaluated through CCK-8, wound healing, Transwell, flow cytometry and xenograft assays. RNA sequencing and rescue experiments with the NF-κB agonist PMA were employed to delineate the underlying mechanism. Results HNRNPA0 was significantly upregulated in OSCC tissues and cell lines, and its high expression predicted poor patient survival. HNRNPA0 knockdown potently suppressed tumour growth both in vitro and in vivo. It inhibited proliferation, migration, invasion, epithelial-mesenchymal transition (EMT) and cell cycle progression, while promoting apoptosis. Transcriptomic analysis revealed a profound impact on the NF-κB signalling pathway. Crucially, activation of this pathway by PMA substantially reversed all anti-tumour phenotypes caused by HNRNPA0 depletion, establishing NF-κB as the critical functional downstream effector. Conclusion Our study identifies HNRNPA0 as a key oncoprotein in OSCC, driving tumour progression primarily via activation of the NF-κB pathway. HNRNPA0 represents a promising prognostic biomarker and a novel therapeutic target for OSCC.

LncRNA RASAL2-AS1 serves as a potential biomarker for prognosis and regulates cellular processes via the miR-590-5p/PDCD5 axis and Wnt/β-catenin pathway in cervical cancer

The objective of this research is to determine the relative abundances and clinical significance of the long non-coding RNA (lncRNA) RASAL2-AS1 in cervical cancer. Additionally, this study aims to elucidate its molecular mechanism in influencing malignant progression through the regulation of the microRNA-590-5p (miR-590-5p)/PDCD5 axis and the Wnt/β-catenin signaling pathway. Gene expression of RASAL2-AS1 was assessed in cervical cancer tissues ( n  = 103) using quantitative real-time PCR (qRT-PCR). The impacts on cellular events, epithelial-mesenchymal transition (EMT), and key signaling pathways were evaluated using CCK-8, Transwell, and Western Blot techniques. The target relationships among RASAL2-AS1, miR-590-5p, and PDCD5 were verified via luciferase reporter assays. RASAL2-AS1 exhibited significantly reduced expression in cervical cancer. Its downregulation was markedly associated with advanced FIGO staging ( P  = 0.014), lymph node metastasis ( P  = 0.035), and poor prognosis, establishing it as an independent indicator of unfavorable outcomes for cervical cancer patients (HR = 0.201, 95% CI = 0.087–0.464, P  < 0.001). Functionally, the upregulation of RASAL2-AS1 inhibited cervical cancer cell growth and EMT. Mechanistically, RASAL2-AS1 acted as a molecular sponge to sequester miR-590-5p, thereby alleviating its inhibitory effect on PDCD5. Furthermore, RASAL2-AS1 obstructs the Wnt/β-catenin pathway through the miR-590-5p/PDCD5 axis. RASAL2-AS1 exerts a tumor-suppressive effect in cervical cancer by functioning as a competing endogenous RNA (ceRNA) that modulates the miR-590-5p/PDCD5 axis and regulates the Wnt/β-catenin pathway.

MiR-326 targets ETS1 to activate PI3K/Akt signaling and promote malignant phenotypes in gastric cancer cells

Abstract: Background: Gastric cancer (GC) remains a leading cause of cancer-related mortality worldwide and effective therapeutic targets remain limited. Objectives: This study aimed to investigate the expression and functional role of miR-326 in gastric cancer, identify its direct target gene and elucidate the underlying molecular mechanisms involving the PI3K/Akt signaling pathway and the regulation of inflammatory cytokines. Methods: miR-326 expression was detected by qRT-PCR in 56 paired GC tissues and adjacent normal tissues, as well as in GC cell lines (SGC-7901, MKN-45, AGS) and normal GES-1 cells. CCK-8 assays, flow cytometry, Transwell migration/invasion assays and ELISA were performed to assess cell proliferation, apoptosis, migration/invasion and cytokine secretion. Dual-luciferase reporter assay and Western blotting were used to validate the target gene and pathway activation. Results: miR-326 was significantly upregulated in GC tissues and cell lines. ETS1 was identified as a direct target of miR-326. Overexpression of miR-326 reduced ETS1 expression, activated the PI3K/Akt pathway (increased p-PI3K and p-AKT), promoted cell proliferation, migration and invasion, inhibited apoptosis and induced an imbalance of inflammatory cytokines (increased IL-6 and TNF-α; decreased IL-10 and IL-17). Co-overexpression of ETS1 or treatment with the PI3K inhibitor LY294002 reversed these effects. Conclusions: miR-326 promotes malignant phenotypes in gastric cancer cells by targeting ETS1, activating the PI3K/Akt signaling pathway, and inducing dysregulation of inflammatory cytokines. The miR-326/ETS1/PI3K/Akt axis may serve as a potential diagnostic biomarker and therapeutic target for gastric cancer.

KLF14 inhibits EMT in esophageal cancer cells through inhibition of the MISP-Wnt/β-catenin signaling axis.

BACKGROUND AND OBJECTIVES Esophageal squamous cell carcinoma (ESCC) dominates esophageal malignancies, and metastasis largely accounts for its dismal prognosis. Epithelial-mesenchymal transition (EMT) is a core driver of tumor metastasis. Mitotic spindle positioning protein (MISP) correlates strongly with EMT, yet its function in ESCC EMT remains uncharacterized. This study explored the role of MISP in ESCC EMT and its molecular regulatory mechanisms. METHODS An in vivo human ESCC cell-derived xenograft model was established by injecting ECA109 cells into the left dorsal flank of nude mice. Cell proliferation was assessed using CCK-8 and colony formation assays, while migration and invasion were evaluated via Transwell assays. The expression of key genes and proteins was analyzed by RT‒qPCR, Western blotting, and immunohistochemistry (IHC). The interaction between KLF14 and MISP was validated through chromatin immunoprecipitation (ChIP) and dual-luciferase reporter gene assays. RESULTS We found that MISP was highly expressed in ESCC samples and that its knockdown significantly suppressed the proliferation, colony formation, migration, and invasion of human ESCC cell lines (KYSE30 and ECA109) while concurrently inhibiting EMT progression (as evidenced by the upregulation of E-cadherin expression and downregulation of N-cadherin/vimentin expression). Furthermore, KLF14 bound to the MISP promoter, was downregulated in ESCC cells, and correlated negatively with MISP levels. Overexpression of KLF14 inhibited tumor growth in nude mouse xenografts and reduced MISP expression and EMT in KYSE30 and ECA109 cells. Additionally, KLF14 overexpression or MISP knockdown in these cells suppressed the expression of Wnt/β-catenin signaling-related proteins (Wnt3a, β-catenin, c-Myc, and cyclin D1). Notably, the EMT phenotype could be partially rescued by either MISP overexpression in KLF14-overexpressing cells or LiCl (a Wnt pathway activator) treatment in MISP-knockdown cells. CONCLUSION Our study demonstrates that KLF14 can inhibit ESCC EMT at least partially via downregulating MISP expression to block downstream Wnt/β-catenin signaling. These findings mechanistically elucidate the role of MISP in promoting ESCC EMT and suggest MISP as a potential therapeutic target for related malignancies.