Qiwei Yang

Colorectal cancer (CRC) metastasis remains a clinically intractable challenge, where the partial epithelial-mesenchymal transition (partial EMT) state is a key driver of metastatic plasticity and treatment resistance. The calcium-activated chloride channel ANO1 (TMEM16A) is frequently overexpressed in epithelial tumors, yet its mechanism in regulating metastasis and partial EMT remains undefined. In this study, we identify ANO1 as a key stabilizer of the partial EMT state. ANO1 was upregulated in CRC tissues and correlated with poor patient survival. Functionally, ANO1 knockdown robustly suppressed CRC cell proliferation, migration, and invasion, and potently induced early apoptosis. Proteomic profiling linked ANO1 depletion to the downregulation of cytoskeletal and adhesion networks. Given the centrality of these processes to EMT, we specifically examined key EMT-associated molecules. Western blot results showed that ANO1 silencing reduced expression of mesenchymal markers (N-cadherin, vimentin, α-SMA) and EMT transcription factors (Zeb1, Snail, Twist), while also downregulating the epithelial marker E-cadherin. Immunofluorescence revealed that ANO1 depletion disrupted a spatially organized hybrid subcellular architecture, and flow cytometry demonstrated that ANO1 knockdown selectively depleted the E-cadherin⁺/vimentin⁺ partial EMT subpopulation. Collectively, these data indicated that ANO1 depletion actively dismantles the organized architecture of the partial EMT state rather than driving a canonical phenotypic reversal. Transcriptomic and integrated multi‑omics analyses further revealed that ANO1 loss induced widespread uncoupling between transcriptional and proteomic regulation, with the mTOR and MAPK pathways enriched among "transcript‑only" changes. Consistently, ANO1 silencing reduced phosphorylation of PI3K, AKT, mTOR, and ERK without altering total protein levels, and pharmacological inhibition of either pathway phenocopied the dissolution of the partial EMT state. In summary, our work delineates a novel mechanism wherein ANO1, acting as an upstream hub, licenses metastatic progression by co‑activating PI3K/AKT/mTOR and ERK signaling to stabilize a plastic, survival‑competent partial EMT state, thereby nominating it as a promising therapeutic target for intercepting CRC dissemination.

The therapeutic efficacy of extracellular vesicles (EVs) depends on the status of their donor cells. Cellular senescence of bone marrow mesenchymal stem ce

The close spatial and temporal connection between osteogenesis and angiogenesis around type H vasculature is referred as “osteogenesis-angiogenesis coupling”, which is one of the basic mechanisms of osteogenesis. Endothelial cells (ECs), bone marrow mesenchymal stem cells (BMSCs), and their specific lineage constitute important cluster that participate in the regulation of osteogenesis and angiogenesis in bone microenvironment. However, the regulatory mechanism of osteogenesis-angiogenesis coupling under the condition of bone healing has not been unveiled. In this study, we demonstrated that the exosome derived from ECs (EC-exo) is an initiator of type H blood vessels formation, and EC-exo acts as a mediator in orchestrating osteogenesis-angiogenesis coupling by enhancing BMSC osteogenic differentiation and EC angiogenesis both in monolayer and stereoscopic co-culture system of primary human cells. The transcriptome array indicated that zinc finger and BTB domain containing 16 (ZBTB16) is a key gene in EC-exo-mediated osteogenesis, and ZBTB16 is indispensable in EC-exo-initiated osteogenesis-angiogenesis coupling. Mechanistically, EC-exo up-regulated the expression of ZBTB16 in BMSCs, thereby promoting osteoprogenitor phenotype transformation; the osteoprogenitors further promote ECs which constitute type H vessel (H-ECs) generation by activating HIF-1α pathway; and the H-ECs conversely promotes osteogenic differentiation of BMSCs. The crosstalk between BMSCs and ECs triggered by EC-exo constitutes a positive feedback loop that enhances osteogenesis-angiogenesis coupling. This study demonstrates that EC-exo can become an effective therapeutic tool to promote bone regeneration and repair.

Introduction: Brain tissue is extremely sensitive to hypoxia/reoxygenation (H/R) injury, which can easily cause irreversible damage to neurons. H/R injury can induce neuronal apoptosis through glutamate-mediated excitotoxicity. N-methyl-d-aspartate receptor (NMDAR) is one of the main receptors of excitatory glutamate, and blocking NMDAR protects brain tissue from ischemic and hypoxic injury. However, NMDAR hypofunction can also cause psychotic symptoms or cognitive impairment. There is still a lack of systematic research on the changes in the proteome and transcriptome in neuronal cells under conditions of NMDAR hypofunction and H/R injury.Methods: We compared the changes in the proteome, transcriptome and lncRNA expression levels in neurons after NMDAR knockdown and H/R by isobaric tags for relative and absolute quantitation (iTRAQ) and RNA sequencing (RNA-Seq).Results: The results showed that the proteins Rps9, Rpl18 and Rpl15 and the lncRNAs XLOC_161072 and XLOC_065271 were significantly downregulated after NMDAR knockdown but upregulated after H/R; in contrast, the mRNAs Bank1 and Pcp4l1 and the lncRNAs XLOC_159404 and XLOC_031922 were significantly upregulated after NMDAR knockdown but downregulated after H/R.Discussion: In this study, we demonstrated the characterization of protein, mRNA, and lncRNA expression profiles in neurons following NMDAR knockdown and H/R injury. These molecules are involved in multiple biological functions and signaling pathways, and their roles in neurons lacking NMDAR and subjected to H/R injury deserve further study. Additionally, we found that lncRNAs respond fastest to hypoxic stimulation and that Gapdh is not suitable as a reference protein for NMDAR-reduced neuron-related experiments.