Angiogenesis related genes based prognostic model of glioma patients developed by multi-omics approach.

INTRODUCTION: Glioma, particularly glioblastoma (GBM), is a highly malignant brain tumor with poor prognosis despite current therapeutic approaches. The tumor microenvironment (TME), plays a crucial role in glioma progression by promoting invasion and drug resistance. Angiogenesis, the formation of new blood vessels, is a tightly regulated process involving endothelial cell activation, proliferation, and migration. In cancer, angiogenesis becomes dysregulated, leading to excessive blood vessel formation. METHODS: We enrolled bulk data of TCGA-LGG/GBM, CGGA-693, and CGGA-325 cohorts, scRNA data of GSE162631, GSE84465, and GSE138794 cohorts. Identification of malignant cells was conducted by "copycat" R package. The "AUCell" R package scored the activity of target gene set of each single cell. Consensus clustering was applied using the "ConsensusClusterPlus" R package, while tumor-infiltrating immune cells were determined using "IOBR" R package. To construct a prognostic model, we used LASSO and multiCOX algorithms based on the expression levels of the 15 hub genes, the efficacy of which was verified by KM and ROC analysis. RESULTS: We identified 4 different malignant cell subclusters in glioma and disclosed their distinct gene expression patterns and interactions within TME. We identified differentially expressed immune-related genes (DE-ARGs) in glioma and found 15 genes that were specifically expressed in the malignant glioma cell populations. Glioma cells with higher expression of these DE-ARGs were associated with gliogenesis, glial cell development, and vasculature development. We found that tumor-infiltrating monocytes were the main interacting cell type within glioma TME. Using the expression patterns of the 15 screened DE-ARGs, we categorized glioma samples into 2 molecular clusters with distinct immune features, suggesting a possible relationship between angiogenesis and immune activation and recruitment. We constructed a prognostic model based on the expression levels of the 15 DE-ARGs and evaluated its predictive ability for glioma patient outcomes, which displayed exceedingly high efficacy. CONCLUSION: We characterized different malignant cell subclusters in glioma and investigate their gene expression patterns and interactions within TME. We constructed a prognostic model based on the expression levels of the 15 DE-ARGs and evaluated its predictive ability for glioma patient outcomes, which displayed exceedingly high efficacy.

Parthenolide inhibits the proliferation and migration of cervical cancer cells via FAK/GSK3ß pathway.

PURPOSE: Cervical cancer (CC) ranks as the fourth most prevalent malignancy among women worldwide, necessitating effective therapeutic interventions to mitigate its detrimental impact on both physical and mental health. Parthenolide (PTL), a natural product of the sesquiterpene lactone derived from Feverfew leaves, has exhibited promising anti-tumor properties in previous studies; however, its precise effects and underlying molecular mechanisms in CC remain elusive. METHODS: In this work, we investigated the effect of PTL on the proliferation and migration of CC cells. Western blot analysis and Reverse transcription­quantitative PCR were used for mechanistic elucidation. RESULTS: Our findings indicated that PTL substantially inhibited the proliferation of HeLa and SiHa CC cell lines in a dose- and time-dependent manner. Moreover, PTL significantly suppressed the migration of CC cells by down-regulating the expression of vascular endothelial growth factor (VEGF), metastasis-associated protein 1 (MTA1), and transforming growth factor-ß1 (TGF-ß1). Mechanistically, PTL blocked the phosphorylation of focal adhesion kinase (FAK) and glycogen synthase kinase-3ß (GSK3ß) induced by epidermal growth factor (EGF). Further investigations revealed that PTL suppressed the proliferation of CC cells by inhibiting the EGF-mediated phosphorylation of the FAK/GSK3ß signaling pathway. CONCLUSION: Taken together, the present in vitro results suggest that PTL may inhibit the proliferation and migration of CC cells through down-regulating the FAK/GSK3ß signaling pathway, providing new insights for the application of PTL in the treatment of CC.

Enzyme-free ratiometric fluorescence and colorimetric dual-signal determination of glyphosate based on copper nanoclusters (ZIF/CuNCs) combined with blue carbon dots (bCDs).

A novel ratio fluorescent and colorimetric dual-signal sensing platform for detecting glyphosate based on blue carbon dots (bCDs) combined with ZIF/CuNCs nanomaterials that encapsulate copper nanoclusters (CuNCs) in a metal-organic framework (MOF). In principle, the immobilization of Cu2+ in ZIF/CuNCs results in complexation with imidazole in ZIF, leading to fluorescence quenching of ZIF/CuNCs, while the reference fluorophore bCDs remains unaffected. In addition, the colorimetric sensing strategy was based on the efficient peroxidase-like activity of bCDs binding to Cu2+, catalyzing H2O2 to generate OH. Under this condition, TMB could be oxidized to form blue oxTMB. However, when glyphosate was involved in the system, the fluorescence of ZIF/CuNCs was restored upon due to the strong chelation between Cu2+ and glyphosate, while the peroxidase-like activity of bCDs/Cu2+ decreased and resulted in the generation of fewer oxTMB, accompanied by a lighter blue color. The sensing platform was successfully applied to the determination of glyphosate in real samples of lake water and cabbage, demonstrating reliable and sensitive performance in practical applications.

Upregulation of miR-200a improves ureteral obstruction-induced renal fibrosis via GAB1/Wnt/ß-catenin signaling.

BACKGROUND: Renal fibrosis is a basic pathological change of almost all chronic kidney disorders. Epithelial-mesenchymal transition (EMT) and excessive extracellular matrix (ECM) accumulation play a crucial role in the process of fibrosis. METHODS: Western blot and qRT-PCR were accomplished to analyze the expression levels of target proteins and genes, respectively. The fibrotic levels in the renal tissues of rats were confirmed utilizing Masson staining. Expression of ECM-related α-SMA in the renal tissues was determined by immunohistochemistry assay. The combination of GRB2 associated binding protein 1 (GAB1) and miR-200a was ensured by starBase database and luciferase reporter assay. RESULTS: Our data uncovered that miR-200a was downregulated, but GAB1 was upregulated in the renal tissues of the rat experienced unilateral ureteral obstruction (UUO). Overexpression of miR-200a improved tissues fibrosis, suppressed GAB1 expression and ECM deposition, and inactivated Wnt/ß-catenin in UUO rats. Moreover, miR-200a expression was inhibited, while GAB1 expression was facilitated in the TGF-ß1-induced HK-2 cells. In TGF-ß1-induced HK-2 cells, miR-200a overexpression inhibited GAB1 expression, also declined ECM-related proteins and mesenchymal markers expression. Oppositely, miR-200a overexpression facilitated epithelial marker expression in the TGF-ß1-induced HK-2 cells. Next, the data revealed that miR-200a inhibited GAB1 expression through binding to the mRNA 3'-UTR of GAB1. Increasing of GAB1 reversed the regulation of miR-200a to GAB1 expression, Wnt/ß-catenin signaling activation, EMT and ECM accumulation. CONCLUSION: Overall, miR-200a increasing improved renal fibrosis through attenuating EMT and ECM accumulation by limiting Wnt/ß-catenin signaling via sponging GAB1, indicating miR-200a may be a promising objective for renal disease therapy.

Harmine exerts anxiolytic effects by regulating neuroinflammation and neuronal plasticity in the basolateral amygdala.

Increasing evidence indicates that an altered immune system is closely linked to the pathophysiology of anxiety disorders, and inhibition of neuroinflammation may represent an effective therapeutic strategy to treat anxiety disorders. Harmine, a beta-carboline alkaloid in various medicinal plants, has been widely reported to display anti-inflammatory and potentially anxiolytic effects. However, the exact underlying mechanisms are not fully understood. Our recent study has demonstrated that dysregulation of neuroplasticity in the basolateral amygdala (BLA) contributes to the pathological processes of inflammation-related anxiety. In this study, using a mouse model of anxiety challenged with Escherichia coli lipopolysaccharide (LPS), we found that harmine alleviated LPS-induced anxiety-like behaviors in mice. Mechanistically, harmine significantly prevented LPS-induced neuroinflammation by suppressing the expression of pro-inflammatory cytokines including IL-1ß and TNF-α. Meanwhile, ex vivo whole-cell slice electrophysiology combined with optogenetics showed that LPS-induced increase of medial prefrontal cortex (mPFC)-driven excitatory but not inhibitory synaptic transmission onto BLA projection neurons, thereby alleviating LPS-induced shift of excitatory/inhibitory balance towards excitation. In addition, harmine attenuated the increased intrinsic neuronal excitability of BLA PNs by reducing the medium after-hyperpolarization. In conclusion, our findings provide new evidence that harmine may exert its anxiolytic effect by downregulating LPS-induced neuroinflammation and restoring the changes in neuronal plasticity in BLA PNs.

Cancer-associated fibroblasts promote malignant phenotypes of prostate cancer cells via autophagy : Cancer-associated fibroblasts promote prostate cancer development.

Dysregulation of autophagy in cancer-associated fibroblasts (CAFs) has been demonstrated to play a role in malignant phenotypes of human tumors. We intended to investigate the function of CAFs autophagy in prostate cancer (PCa). Firstly, CAFs and normal fibroblasts (NFs) were isolated from cancerous and adjacent normal tissues of PCa patients, for the following experimental preparation. In comparison with NFs, CAFs expressed higher levels of the myofibroblast marker ?-smooth muscle actin (?-SMA) and the mesenchymal marker Vimentin. Besides, CAFs possessed a higher autophagic level than NFs. As for malignant phenotypes, PCa cells co-cultured with CAFs-CM showed greater proliferation, migration and invasion capabilities, while these outcomes were obviously abolished by autophagy inhibition with 3-Methyladenine (3-MA). Moreover, silencing of ATG5 in CAFs inhibited fibroblasts autophagic level and suppressed malignant phenotypes of PCa cells, while ATG5 overexpression in NFs exerted opposite effects. Depletion of ATG5 in CAFs inhibited the xenograft tumor growth and lung metastasis of PCa cells. Taken together, our data demonstrated the promotive effect of CAFs on PCa malignant phenotypes through ATG5-dependent autophagy, suggesting a novel mechanism for PCa progression.

Identification of Prognostic Markers of DNA Damage and Oxidative Stress in Diagnosing Papillary Renal Cell Carcinoma Based on High-Throughput Bioinformatics Screening.

Purpose: Papillary renal cell carcinoma (pRCC) is the second most common histological subtype of adult kidney tumors, with a poor prognosis due to limited understanding of the disease mechanism. Herein, we have performed high-throughput bioinformatic screening to explore and identify potential biomarkers of DNA damage and oxidative stress for pRCC. Methods: RNA sequencing data related to pRCC were downloaded from the TCGA database, and differentially expressed genes (DEG) were identified by a wide variety of clustering and classification algorithms, including self-organized maps (SOM), artificial neural networks (ANN), support vector machines (SVM), fuzzy logic, and hyphenated techniques such as neuro-fuzzy networks. Then DAVID and STRING online biological information tools were used to analyze functional enrichment of the regulatory networks of DEG and construct a protein-protein interaction (PPI) network, and then the Cytoscape software was used to identify hub genes. The importance of key genes was assessed by the analysis of the Kaplan-Meier survival curves using the R software. Lastly, we have analyzed the expression of hub genes of DNA damage and oxidative stress (BDKRB1, NMUR2, PMCH, and SAA1) in pRCC tissues and adjacent normal tissues, as well as the relationship between the expression of hub genes in pRCC tissues and pathological characteristics and prognosis of pRCC patients. Results: A total of 1,992 DEGs for pRCC were identified, with 1,142 upregulated ones and 850 downregulated ones. The DEGs were significantly enriched in activities including DNA damage and oxidative stress, chemical synaptic transmission, an integral component of the membrane, calcium ion binding, and neuroactive ligand-receptor interaction. cytoHubba in the Cytoscape software was used to determine the top 10 hub genes in the PPI network as BDKRB2, NMUR2, NMU, BDKRB1, LPAR5, KNG1, LPAR3, SAA1, MCHR1, PMCH, and NCAPH. Furthermore, the expression level of hub genes BDKRB1, NMUR2, PMCH, and SAA1 in pRCC tissues was significantly higher than that in the adjacent normal tissues. Meanwhile, the expression level of hub genes BDKRB1, NMUR2, PMCH, and SAA1 in pRCC tissues was significantly positively correlated with tumor stage, lymph node metastasis, and the histopathology grade of pRCC. In addition, high expression levels of hub genes BDKRB1, NMUR2, PMCH, and SAA1 were associated with a poor prognosis for patients with pRCC. Univariate and multivariate analyses showed that the expression of hub genes BDKRB1, NMUR2, PMCH, and SAA1 were independent risk factors for the prognosis of patients with pRCC. Conclusion: The results of this analysis suggested that BDKRB1, NMUR2, PMCH, and SAA1 might be potential prognostic biomarkers and novel therapeutic targets for pRCC.