Background Laryngeal squamous cell carcinoma (LSCC) is a common malignant tumor in otorhinolaryngology, with a high incidence rate and a tendency of lymph node metastasis. This study aims to systematically evaluate the expression of the transcription factor SOX4 in LSCC and its clinical relevance, clarify its impact on the biological behavior of tumors and potential molecular mechanisms, and provide a theoretical basis for diagnosis and treatment. Method Bioinformatics analysis was conducted based on the GEO dataset (GSE201777) to evaluate the expression of SOX4 and its clinical relevance; The expression of SOX4 in LSCC clinical tissues and cell lines was verified by qRT-PCR. SOX4 knockdown and overexpression cell models were constructed. Proliferation was evaluated by MTS and colony formation assays, migration and invasion were evaluated by scratch and Transwell assays, and apoptosis was detected by flow cytometry and TUNEL. The downstream effector molecules of SOX4 were screened by transcriptome sequencing and verified by functional recovery experiments. The association between SOX4 and tumor immune infiltration as well as drug sensitivity was explored by combining bioinformatics analysis. Ultimately, the effects of SOX4 on tumor growth and lymph node metastasis in vivo were evaluated in a xenograft mouse model. Result Bioinformatics and experimental data consistently showed that SOX4 was upregulated in LSCC and positively correlated with disease progression and lymph node metastasis. In vitro experiments have shown that knockdown of SOX4 significantly inhibits the proliferation, migration and invasion of LSCC cells, while overexpression of SOX4 promotes a malignant phenotype. Transcriptome and functional verification determined that PTBP2 is a downstream effector molecule of SOX4. The restored expression of PTBP2 can partially alleviate the inhibitory effect caused by SOX4 knockdown. Bioinformatics analysis further suggests that the SOX4/PTBP2 axis is associated with the tumor immune microenvironment and the sensitivity to several anti-cancer drugs. In vivo studies have shown that knockdown of SOX4 or administration of erlotinib significantly inhibited tumor growth and reduced the rate of lymph node metastasis. Conclusion SOX4 promotes the growth and lymph node metastasis of LSCC by regulating PTBP2. The SOX4-PTBP2 axis may become a potential diagnostic and therapeutic target for LSCC.

The full text of this preprint has been withdrawn by the authors while they make corrections to the work. Therefore, the authors do not wish this work to be cited as a reference. Questions should be directed to the corresponding author.

Diabetic peripheral neuropathy (DPN) is a common chronic complication of diabetes mellitus, and Schwann cell dysfunction contributes to its pathogenesis and progression. IGF2BP2 is a single-stranded RNA-binding protein that has recently been identified as an m6A reader; however, its role and regulatory effects in Schwann cells during DPN remain largely unknown. In this study, we demonstrated that high-glucose exposure significantly increased IGF2BP2 levels in RSC96 cells and in the sciatic nerves of both type 1 and type 2 diabetic mice, accompanied by reduced autophagy and neurotrophin expression. Consistently, IGF2BP2 expression was associated with metabolic and immune-related parameters in patients with type 2 diabetes. Functionally, IGF2BP2 overexpression inhibited, whereas its downregulation partially rescued, high glucose-induced suppression of autophagy and neurotrophin expression in vitro (RSC96 cells) and in vivo (IGF2BP2 knockout mice). Mechanistically, IGF2BP2 bound to EGR1 mRNA in an m6A modification-dependent manner, primarily recognizing an adenosine at position 1268, thereby enhancing EGR1 mRNA stability. Downregulation of EGR1 alleviated the inhibitory effects of IGF2BP2 overexpression on autophagy and neurotrophin expression in RSC96 cells. Conversely, EGR1 overexpression attenuated the protective effects of IGF2BP2 knockout on peripheral nerve function and neurotrophic signaling in diabetic mice. Overall, high-glucose stimulation upregulated IGF2BP2, which enhanced EGR1 mRNA stability in an m6A-dependent manner, thereby suppressing autophagy and neurotrophin expression in Schwann cells during DPN.

Febrile seizure is a common pediatric neurological emergency that may potentially increase the risks of epilepsy and neurodevelopmental disorders. Ginsenosides are the primary active component of ginseng, demonstrating notable neuroprotective effects. However, the effects and mechanisms of Ginsenosides in febrile seizures remain poorly understood. This study aims to investigate the effects and potential molecular targets of Ginsenosides in mitigating febrile seizures and further elucidate its underlying mechanism. Seizure behaviors and EEG recordings were conducted in mice to investigate the effect of Ginsenosides on febrile seizures. Direct targets of Ginsenosides were identified through Thermal Proteome Profiling (TPP), subsequently validated by Surface Plasmon Resonance (SPR), Cellular Thermal Shift Assay (CETSA), and molecular docking. The mechanisms of Ginsenosides targeting Dynamin-related protein 1 (Drp1) in inhibiting febrile seizures were elucidated through proteomic analysis, molecular biology techniques, mitochondrial function assessments, and metabolite profiling. These findings demonstrate that Ginsenosides significantly attenuated febrile seizure severity and reduced the incidence of generalized tonic-clonic seizures (GTCS), showing a superior safety and efficacy profile compared to Ilepcimide. Using the TPP method, we identified and validated Drp1 as a promising direct target for therapeutic intervention in febrile seizures. Mechanistically, Ginsenoside-mediated Drp1 inhibition restored mitochondrial calcium homeostasis, promoting ATP production and elevating region-specific cortical levels of the endogenous anti-seizure metabolite adenosine. Moreover, Ginsenosides upregulated adenosine A1 receptor expression and suppressed the cyclic Adenosine Monophosphate (cAMP) signaling pathway, ultimately exerting anti-seizure effects. In summary, this study reveals that Ginsenosides significantly inhibit febrile seizures by directly targeting Drp1, thereby increasing adenosine levels and suppressing cAMP signaling, effectively suppressing seizures. Our findings demonstrate the potential of Ginsenosides in febrile seizures prevention and highlight Drp1 as a promising therapeutic target, thereby providing novel strategies for identifying targets of bioactive compounds.

BackgroundLaryngeal squamous cell carcinoma (LSCC) is a common malignant tumor in otorhinolaryngology, with a high incidence rate and a tendency of lymph node metastasis. This study aims to systematically evaluate the expression of the transcription factor SOX4 in LSCC and its clinical relevance, clarify its impact on the biological behavior of tumors and potential molecular mechanisms, and provide a theoretical basis for diagnosis and treatment.MethodBioinformatics analysis was conducted based on the GEO dataset (GSE201777) to evaluate the expression of SOX4 and its clinical relevance; The expression of SOX4 in LSCC clinical tissues and cell lines was verified by qRT-PCR. SOX4 knockdown and overexpression cell models were constructed. Proliferation was evaluated by MTS and colony formation assays, migration and invasion were evaluated by scratch and Transwell assays, and apoptosis was detected by flow cytometry and TUNEL. The downstream effector molecules of SOX4 were screened by transcriptome sequencing and verified by functional recovery experiments. The association between SOX4 and tumor immune infiltration as well as drug sensitivity was explored by combining bioinformatics analysis. Ultimately, the effects of SOX4 on tumor growth and lymph node metastasis in vivo were evaluated in a xenograft mouse model.ResultBioinformatics and experimental data consistently showed that SOX4 was upregulated in LSCC and positively correlated with disease progression and lymph node metastasis. In vitro experiments have shown that knockdown of SOX4 significantly inhibits the proliferation, migration and invasion of LSCC cells, while overexpression of SOX4 promotes a malignant phenotype. Transcriptome and functional verification determined that PTBP2 is a downstream effector molecule of SOX4. The restored expression of PTBP2 can partially alleviate the inhibitory effect caused by SOX4 knockdown. Bioinformatics analysis further suggests that the SOX4/PTBP2 axis is associated with the tumor immune microenvironment and the sensitivity to several anti-cancer drugs. In vivo studies have shown that knockdown of SOX4 or administration of erlotinib significantly inhibited tumor growth and reduced the rate of lymph node metastasis.ConclusionSOX4 promotes the growth and lymph node metastasis of LSCC by regulating PTBP2. The SOX4-PTBP2 axis may become a potential diagnostic and therapeutic target for LSCC.

Schwann cells dysfunction is a key contributor to diabetic peripheral neuropathy (DPN), affecting both neurons and blood vessels. However, the precise mechanisms underlying high glucose-induced Schwann cells dysfunction are still not fully elucidated. In the present study, we investigated the expression, function and molecular mechanisms of SDF2L1 in Schwann cells using diabetic mice, SDF2L1 KO mice, rat Schwann cell (RSC96) and primary rat Schwann cell (PRSC). The RNA-seq of high glucose-treated RSC96 cells revealed an evident downregulation of SDF2L1 at both 48 and 72 h. The inhibition of high glucose on SDF2L1 expression was further confirmed at the levels of mRNA and protein in RSC96 and PRSC cells. Again, reduced SDF2L1 expression was also observed in the sciatic nerves of both type 1 and 2 diabetic mice. Functional exploration revealed that SDF2L1 knockdown in RSC96 cells suppressed the expression of LC3-II, P62, BDNF, NGF and IGF. In vivo SDF2L1 KO also decreased these proteins expression in the sciatic nerve of C57BL/6 J mice, along with the reduced nerve conduction velocity and action potential amplitude. Then, proteomics analyses and biological experiments demonstrated that SDF2L1 knockdown significantly decreased KPNA3 expression in RSC96 cells. Overexpression of KPNA3 ameliorated the decreases in LC3-II, P62, BDNF, NGF and IGF caused by SDF2L1 downregulation in vitro. Moreover, KPNA3 affected the nuclear import of transcription factors TFEB and CREB in RSC96 cells. Next, KPNA3 overexpression reversed SDF2L1 KO-reduced the nuclear aggregation of TFEB and CREB, and the expression of LC3, P62, BDNF and NGF in vivo. Collectively, these findings suggest that decreased SDF2L1 inhibits cell autophagy and neurotrophin expression by impeding the nuclear import of TFEB and CREB via KPNA3 downregulation in high glucose-treated Schwann cells.

Soy isoflavones have been reported to inhibit breast cancer (BC) progression; however, the underlying mechanism remains unclear. Previous studies have primarily focused on individual components of soy isoflavones. This study integrates network pharmacology, molecular docking, molecular dynamics simulations, experimental validation, and Mendelian randomization (MR) to systematically evaluate the effects and mechanisms of soy isoflavones on BC. A total of 190 potential anti-BC targets for six active soy isoflavone components were identified from multiple public databases. Enrichment analysis revealed that these targets play critical roles in regulating cellular oxidative stress and modulating drug sensitivity in BC. Ten hub targets were identified through protein-protein interaction (PPI) network analysis and topology screening: TP53, SRC, ESR1, EGFR, PIK3CA, HSP90AA1, PRKACA, HRAS, AKT1, and ITGB1. Molecular docking analysis demonstrated strong binding between these hub targets and the six soy isoflavone components, with PRKACA-daidzin (DA) and PRKACA-genistin (GE) exhibiting the strongest binding affinities. Molecular dynamics simulations further confirmed the stability of the binding interactions of these two complexes. Experimental validation indicated that DA and GE effectively inhibited BC progression, with their mechanism linked to the suppression of PRKACA expression. However, MR analyses did not find a causal relationship between the consumption of soy products and reduced BC risk. In conclusion, this study confirms the anti-BC potential of soy isoflavones and, for the first time, elucidates the anti-BC mechanism of soy isoflavones.

F-box and WD repeat domain-containing 7 (FBXW7) protein is known as one of the crucial components of the E3 ubiquitin ligase called the Skp1-Cullin1-F-box (SCF) complex, which regulates the degradation of a network of proteins via the ubiquitin-proteasome system. In our study, we investigated the latent impact of FBXW7 on renal tubular cells injury and its molecular mechanism in diabetic kidney disease (DKD). FBXW7 was upregulated in kidneys of diabetic mice and human renal proximal tubular cells exposed to high glucose. Again, the function of experiment found that overexpression of FBXW7 led to epithelial-mesenchymal transition (EMT) of HK2 cells, as indicated by decreased E-cadherin and increased α-smooth muscle actin (α-SMA). Knockdown of FBXW7 ameliorated high glucose-induced EMT of HK2 cells via downregulation of TGF-β1. Then, FBXW7 overexpression downregulated the stability of the KLF5 protein and promoted protein ubiquitination in normal glucose-cultured HK2 cells, which was significantly reversed by the addition of MG132, a specific proteasome inhibitor. Furthermore, overexpression of KLF5 effectively prevented FBXW7 upregulation-induced EMT in HK2 cells. Finally, chemical inhibitors or mTOR kinase dead vector to interfere the activity of mTOR effectively suppressed FBXW7 expression in HK2 cells treated with high glucose. Taken together, these above data suggest that mTOR signaling pathway-regulated FBXW7 mediates high glucose-induced EMT of renal tubular cells by affecting the stability of KLF5.

ObjectiveOur previous study found that Achaete-scute complex homolog 1 (ASCL1) is involved in classifying BC subtypes with different prognostic and pathological characteristics. However, the biological role of ASCL1 in BC still remains largely unexplored. This study aims to elucidate the function of ASCL1 in BC using bioinformatics analyses, as well as in vitro and in vivo experimental approaches.MethodsData from the TCGA, GEO, and Human Protein Atlas databases were utilized to evaluate ASCL1 expression in BC and its association with patient prognosis. Genetic alterations in ASCL1 were assessed through the COSMIC and cBioPortal databases, while the TIMER2.0 database provided insights into the relationship between ASCL1 expression and key gene mutations in BC. The GDSC database was used to examine correlations between ASCL1 levels and sensitivity to standard chemotherapeutic agents. Associations between ASCL1 expression and cytokines, immunomodulatory factors, MHC molecules, and receptors were analyzed using Pearson and Spearman correlation methods. The TIP database was employed to investigate the connection between ASCL1 expression and immunoreactivity scores, and six computational approaches were applied to evaluate immune cell infiltration. Functional assays were conducted on BC cell lines MCF-7 and MDA-MB-231, and nude mouse models were used for in vivo studies.ResultsASCL1 was found to be upregulated in BC and correlated with unfavorable prognosis and mutations in key oncogenes. Its expression was linked to immunomodulatory factors, immune cell infiltration, and immunoreactivity scores in the tumor microenvironment. Additionally, ASCL1 influenced tumor immune dynamics and chemosensitivity in BC. Overexpression of ASCL1 enhanced BC cell proliferation, migration and invasion, while its knockdown had the opposite effect. Notably, inhibition of ASCL1 increased BC cell sensitivity to paclitaxel both in vitro and in vivo. In addition, inhibition of ASCL1 activated ferroptosis in BC, including altered mitochondrial morphology, increased MDA and ROS levels, decreased GSH levels and reduced GSH/GSSG ratio. Mechanistically, inhibition of ASCL1 decreases the phosphorylation of CREB1, thus reducing the expression of GPX4. In summary, inhibition of ASCL1 increases paclitaxel sensitivity by activating ferroptosis via the CREB1/GPX4 axis.ConclusionsASCL1 exerts oncogenic effects in BC and represents a potential therapeutic target for intervention.

Diabetic kidney disease (DKD) is a prevalent complication associated with diabetes in which podocyte dysfunction significantly contributes to the development and progression of the condition. Ring finger protein 183 (RNF183) is an ER-localized, transmembrane ring finger protein with classical E3 ligase activity. However, whether RNF183 is involved in glomerular podocyte dysfunction, which is the mechanism of action of DKD, is still poorly understood. In this study, we first demonstrated that RNF183 expression in glomerular podocytes of patients with DKD decreased as the disease progressed. Additionally, our transcriptome sequencing analysis of kidney tissues from diabetic mice revealed a significant reduction in RNF183 expression within the kidney cortex. Similarly, the expression of RNF183 was significantly reduced both in the kidneys of diabetic mice and in human podocytes exposed to high glucose conditions. The downregulation of RNF183 resulted in a suppression of autophagic activity, an increase in apoptotic cell death, and reduced expression of cellular markers in HPC cells. We found that RNF183 was modified via N6-methyladenosine (m6A) RNA methylation. Meanwhile, treatment with meclofenamic acid 2 (MA2), an m6A demethylase inhibitor, resulted in the upregulation of RNF183 expression in HPC cells cultured in high glucose conditions. Furthermore, high glucose treatment decreased the transcription and protein levels in both the m6A writer methyltransferaselike3 (METTL3) and the m6A reader insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2). IGF2BP2 assisted with METTL3, which is jointly involved in the transcription of RNF183. Furthermore, we confirmed that RNF183 directly ubiquitinates M2 pyruvate kinase (PKM2) through co-immunoprecipitation (Co-IP) and liquid chromatography-mass spectrometry (LC-MS) experiments. The level of PKM2 ubiquitination was increased following RNF183 overexpression, leading to enhanced PKM2 protein degradation and subsequently alleviating high glucose-induced podocyte damage. The results of this study indicated that RNF183 was regulated via m6A methylation modification and that RNF183 expression was reduced in HPC cells treated with high glucose, which resulted in decreased PKM2 ubiquitination levels and subsequently aggravated podocyte injury. The findings suggest that RNF183 may serve as a potential therapeutic target for diabetic kidney injury, offering new insights into its role in the progression of DKD.

Dysfunction of Schwann cells, including cell apoptosis, autophagy inhibition, dedifferentiation, and pyroptosis, is a pivotal pathogenic factor in induced diabetic peripheral neuropathy (DPN). Histone deacetylases (HDACs) are an important family of proteins that epigenetically regulate gene transcription by affecting chromatin dynamics. Here, we explored the effect of HDAC1 on high glucose-cultured Schwann cells. HDAC1 expression was increased in diabetic mice and high glucose-cultured RSC96 cells, accompanied by cell apoptosis. High glucose also increased the mitochondrial pathway apoptosis-related Bax/Bcl-2 and cleaved caspase-9/caspase-9 ratios and decreased endoplasmic reticulum response-related GRP78, CHOP, and ATF4 expression in RSC96 cells (P < 0.05). Furthermore, overexpression of HDAC1 increased the ratios of Bax/Bcl-2, cleaved caspase-9/caspase-9, and cleaved caspase-3 and reduced the levels of GRP78, CHOP, and ATF4 in RSC96 cells (P < 0.05). In contrast, knockdown of HDAC1 inhibited high glucose-promoted mitochondrial pathway apoptosis and suppressed the endoplasmic reticulum response. Moreover, RNA sequencing revealed that U4 spliceosomal RNA was significantly reduced in HDAC1-overexpressing RSC96 cells. Silencing of U4 spliceosomal RNA led to an increase in Bax/Bcl-2 and cleaved caspase-9 and a decrease in CHOP and ATF4. Conversely, overexpression of U4 spliceosomal RNA blocked HDAC1-promoted mitochondrial pathway apoptosis and inhibited the endoplasmic reticulum response. In addition, alternative splicing analysis of HDAC1-overexpressing RSC96 cells showed that significantly differential intron retention (IR) of Rpl21, Cdc34, and Mtmr11 might be dominant downstream targets that mediate U4 deficiency-induced Schwann cell dysfunction. Taken together, these findings indicate that HDAC1 promotes mitochondrial pathway-mediated apoptosis and inhibits the endoplasmic reticulum stress response in high glucose-cultured Schwann cells by decreasing the U4 spliceosomal RNA/IR of Rpl21, Cdc34, and Mtmr11.

Objective The aim of this study was to identify the molecular subtypes of breast cancer based on chromatin regulator-related genes. Methods The RNA sequencing data of The Cancer Genome Atlas-Breast Cancer cohort were obtained from the official website, while the single-cell data were downloaded from the Gene Expression Omnibus database (GSE176078). Validation was performed using the Molecular Taxonomy of Breast Cancer International Consortium dataset. Furthermore, the immune characteristics, tumor stemness, heterogeneity, and clinical characteristics of these molecular subtypes were analyzed. The correlation between chromatin regulators and chemotherapy resistance was examined in vitro using the quantitative real-time polymerase chain reaction (qRT-PCR) and Cell Counting Kit-8 (CCK8) assays. Results This study identified three stable molecular subtypes with different prognostic and pathological features. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and protein-protein interaction analyses revealed that the differentially expressed genes were associated with disease processes, such as mitotic nuclear division, chromosome segregation, condensed chromosome, and specific chromosome region. The T stage and subtypes were correlated with the clinical features. Tumor heterogeneity (mutant-allele tumor heterogeneity, tumor mutational burden, purity, and homologous recombination deficiency) and tumor stemness (RNA expression-based stemness score, epigenetically regulated RNA expression-based stemness score, DNA methylation-based stemness score, and epigenetically regulated DNA methylation-based stemness score) significantly varied between the three subtypes. Furthermore, Western blotting, qRT-PCR, and CCK8 assays demonstrated that the expression of ASCL1 was positively correlated with chemotherapy resistance in breast cancer. Conclusion This study identified the subtypes of breast cancer based on chromatin regulators and analyzed their clinical features, gene mutation status, immunophenotype, and drug sensitivity. The results of this study provide effective strategies for assessing clinical prognosis and developing personalized treatment strategies.

OBJECTIVE:Rab11A is an important molecule for recycling endosomes and is closely related to the proliferation, invasion, and metastasis of tumors. This study investigated the prognostic and immune significance of Rab11A and validated its potential function and mechanism in breast cancer (BRCA). METHODS:RNA sequencing data for 33 tumors were downloaded from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression databases. Correlation analysis was used to evaluate the relationship between Rab11A expression and immune characteristics. Potential pathways were identified using the Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analysis. Immunohistochemical analysis, colony formation assay, bromodeoxyuridine incorporation assay, immunofluorescence, and Western blot were used to explore potential function and mechanism. RESULTS:Analysis of the TCGA database showed significant upregulation of Rab11A expression in a variety of cancers. Rab11A was up-regulated in 82.4% of BRCA. High Rab11A expression is associated with poor survival in cancer patients and is a predictor of poor prognosis. CIBERSORT analysis showed that Rab11A was negatively associated with almost all immune cycle activity scores pan-cancer. The results of the TCGA-BRCA cohort were further confirmed by using pathological samples from clinical BRCA patients. The results showed that Rab11A expression was correlated with estrogen receptor (ER) and progesterone receptor expression in BRCA (p < 0.05). Knockdown and overexpression of Rab11A affected the proliferation of BRCA cells. Further mechanistic studies revealed that down-regulation of ER alpha (ERα) and up-regulation of ER beta (ERβ) mediated Rab11A-induced inhibition of BRCA cell proliferation. CONCLUSION:Rab11A expression in pan-cancer is associated with poor prognosis and immune profile. In particular, in BRCA, Rab11A expression regulates cell proliferation by targeting ERα and ERβ. High Rab11A expression is tightly associated with immune characteristics, tumor microenvironment, and genetic mutations. These results provide a reference for exploring the role of Rab11A in pan-cancer and provide a new perspective for revealing potential therapeutic targets in BRCA.

Acute kidney injury (AKI) is a prevalent and potentially life-threatening complication characterized by a high incidence and mortality. A large number of studies have emphasized the role of ferroptosis in AKI. Moreover, FBXW7, a ubiquitin ligase, has been implicated in acute organ injury. Analysis of the GEO database (GSE98622) revealed increased FBXW7 mRNA levels in the kidney following ischemia‒reperfusion (IR). However, the role of FBXW7 in AKI has not been elucidated. Therefore, this study aimed to investigate the role of FBXW7 in IR-AKI and its underlying mechanisms. Here, we found that IR could induce AKI and increase FBXW7 expression, while the ferroptosis inhibitor Fer-1 alleviated AKI and decreased FBXW7 expression. Furthermore, we treated HK-2 cells with hypoxia for 12 h and reoxygenation for 4 h (H12R4) to simulate IR-AKI and investigated the impact of modulating FBXW7 expression on ferroptosis by employing ferroptosis-related agonists or inhibitors. Our findings revealed that H12R4 induced HK2 ferroptosis and increased the expression of FBXW7. FBXW7 overexpression in control cells exacerbated erastin-induced ferroptosis, and FBXW7 knockdown inhibited ferroptosis in H12R4-treated cells. Mechanistically, we confirmed that FBXW7 can bind to GPX4, a key molecule that inhibits ferroptosis. The half-life of the GPX4 protein decreased after FBXW7 overexpression, GPX4 ubiquitination increased after H12R4, and GPX4 degradation decreased after FBXW7 knockdown. In conclusion, our results indicated that FBXW7 plays an important role in the development of IR-AKI by promoting ferroptosis through the downregulation of GPX4 expression. This study provides new insight into FBXW7 as a potential target for treating AKI.

Background Evidence suggests that enolase-phosphatase 1 (ENOPH1) is involved in the progression of some certain types of cancers and acts as an oncogenic factor in tumor progression. The present study aimed to identify the central role of ENOPH1 in the progression of breast cancer (BC), a highly proliferative and aggressive disease. Methods and results ENOPH1 expression in BC tissues was explored based on the online resource and 40 paired fresh BC and para-carcinoma samples. Functional assays were performed to evaluate the biological effect of ENOPH1 on cell proliferation and migration in ENOPH1-silenced or overexpressing BC cell lines. Blockade of NF-κB by BAY11-7082 was performed to evaluate whether ENOPH1 exerted tumor-promoting properties via regulating the NF-κB signaling pathway. Results of the present study demonstrated that ENOPH1 expression was profoundly upregulated in BC tissues compared with adjacent breast tissues, and ENOPH1 expression was associated with cancer stage, node metastasis status, and overall survival. Functional assays demonstrated that ENOPH1 overexpression significantly accelerated BC cell proliferation, migration, and invasion, while genetic knockdown of ENOPH1 yielded the opposite effects. Mechanistically, ENOPH1 activated the NF-κB pathway, as evidenced by increased expression of NF-κB downstream genes and enhanced NF-κB p65 nuclear translocation. Furthermore, the oncogenic properties of ENOPH1 in proliferation, migration, and invasion were restrained following inhibition of the NF-κB signaling pathway. Conclusions These findings indicated the significance of ENOPH1 in promoting cell proliferation and invasion, mainly through activating the NF-κB pathway, suggesting that ENOPH1 might be an attractive prognostic factor and a potential target for BC therapy.

Diabetic kidney disease is a common complication of diabetes and remains the primary cause of end-stage kidney disease in the general population. Schisandrin B (Sch B) is an active ingredient in Schisandra chinensis. Our study illustrates that Sch B can mitigate renal tubular cell (RTC) epithelial–mesenchymal transition (EMT) and mitochondrial dysfunction in db/db mice, accompanied by the downregulation of TGF-β1 and the upregulation of PGC-1α. Similarly, Sch B demonstrated a protective effect by reducing the expression of TGF-β1, α-SMA, fibronectin, and Col I, meanwhile enhancing the expression of E-cadherin in human RTCs (HK2 cells) stimulated with high glucose. Moreover, under high glucose conditions, Sch B effectively increased mitochondrial membrane potential, lowered ROS production, and increased the ATP content in HK2 cells, accompanied by the upregulation of PGC-1α, TFAM, MFN1, and MFN2. Mechanistically, the RNA-seq results showed a significant increase in KCP mRNA levels in HK2 cells treated with Sch B in a high glucose culture. The influence of Sch B on KCP mRNA levels was confirmed by real-time PCR in high glucose-treated HK2 cells. Depletion of the KCP gene reversed the impact of Sch B on TGF-β1 and PGC-1α in HK2 cells with high glucose level exposure, whereas overexpression of the KCP gene blocked EMT and mitochondrial dysfunction. Furthermore, the PI3K/Akt pathway was inhibited and the AMPK pathway was activated in HK2 cells exposed to a high concentration of glucose after the Sch B treatment. Treatment with the PI3K/Akt pathway agonist insulin and the AMPK pathway antagonist compound C attenuated the Sch B-induced KCP expression in HK2 cells exposed to a high level of glucose. Finally, molecular autodock experiments illustrated that Sch B could bind to Akt and AMPK. In summary, our findings suggested that Sch B could alleviate RTC EMT and mitochondrial dysfunction by upregulating KCP via inhibiting the Akt pathway and activating the AMPK pathway in DKD.

Diabetic encephalopathy (DE) is one of the complications of diabetes mellitus with mild-to-moderate cognitive impairment. Trichostatin A (TSA) has been revealed to show protective effect on central nervous systems in Alzheimer’s disease (AD) and hypoxic–ischemic brain injury. However, the effect and molecular mechanism of TSA on cognitive function of DE are unknown. Here, we demonstrated that cognitive function was damaged in diabetic mice versus normal mice and treatment with TSA improved cognitive function in diabetic mice. Proteomic analysis of the hippocampus revealed 174 differentially expressed proteins in diabetic mice compared with normal mice. TSA treatment reversed the expression levels of 111 differentially expressed proteins grouped into functional clusters, including the longevity regulating pathway, the insulin signaling pathway, peroxisomes, protein processing in the endoplasmic reticulum, and ribosomes. Furthermore, protein–protein interaction network analysis of TSA-reversed proteins revealed that UBA52, CAT, RPL29, RPL35A, CANX, RPL37, and PRKAA2 were the main hub proteins. Multiple KEGG pathway-enriched CAT and PRKAA2 levels were significantly decreased in the hippocampus of diabetic mice versus normal mice, which was reversed by TSA administration. Finally, screening for potential similar or ancillary drugs for TSA treatment indicated that HDAC inhibitors ISOX, apicidin, and panobinostat were the most promising similar drugs, and the PI3K inhibitor GSK-1059615, the Aurora kinase inhibitor alisertib, and the nucleophosmin inhibitor avrainvillamide-analog-6 were the most promising ancillary drugs. In conclusion, our study revealed that CAT and PRKAA2 were the key proteins involved in the improvement of DE after TSA treatment. ISOX, apicidin, and panobinostat were promising similar drugs and that GSK-1059615, alisertib, and avrainvillamide-analog-6 were promising ancillary drugs to TSA in the treatment of DE.

Diabetic peripheral neuropathy (DPN) is the most common complication of diabetes mellitus and no effective therapy is approved. Here, lycorine, a natural alkaloid, was identified as a potential drug for DPN by the bioinformatics analysis of GEO datasets and Connectivity Map database. Lycorine administration improved peripheral nerve function and autophagy-associated proteins of diabetic mice. Again, in vitro high glucose-cultured rat Schwann cells (RSC96) showed enhanced autophagosome marker LC3-II with the treatment of lycorine. Additionally, beclin-1 and Atg3 were decreased in high glucose-stimulated RSC96 cells, which were reversed by lycorine treatment. Furthermore, DPN-associated differentially expressed genes (DEGs) from GEO datasets and lycorine-drug targets from PubChem and PharmMapper were visually analyzed and revealed that MMP9 was both DPN-associated DEGs and lycorine-drug target. Functional enrichment analysis of MMP9-relevant genes showed that cell energy metabolism was involved. Moreover, lycorine reduced high glucose-enhanced MMP9 expression in RSC96 cells. Overexpression of MMP9 attenuated lycorine-induced the expression of beclin-1, Atg3 and LC3-II in high glucose-cultured RSC96 cells. In addition, AMPK pathway activation was confirmed in lycorine-treated high glucose-cultured RSC96 cells. Then AMPK pathway inhibition attenuated lycorine-reduced MMP9 expression in high glucose-treated RSC96 cells. Molecular docking analysis revealed that lycorine bound the domain of AMPK containing Thr 172 site, which affected AMPK (Thr 172) phosphorylation. Finally, AMPK pathway activation and MMP9 downregulation were also revealed in the sciatic nerves of diabetic mice administrated with lycorine. Taken together, lycorine was advised to promote Schwann cell autophagy via AMPK pathway activation and MMP9 downregulation-induced LC3-II transformation in diabetic peripheral neuropathy.

Proliferative vitreoretinopathy (PVR) is a serious ophthalmic disease and characterized by the formation of proliferative membranes by retinal pigment epithelial (RPE) cells. In PVR, the contraction and traction of the fibrocellular membranes cause retinal detachment, which can cause reduction surgery for retinal detachment to fail. Fibroblast growth factor-2 (FGF-2) causes RPE cells to form extracellular matrix (ECM), promotes chemotaxis, mitosis, and positively promotes the disease process of PVR. Plumbagin (PLB) is a plant small molecule naphthoquinone compound. It has the functions in anti-tumor, anti-inflammatory, inhibit proliferation. We tried to investigate the possible effects of PLB on the biological behavior of ARPE-19 cells induced by FGF-2 and its underlying mechanisms. Our study confirmed that proliferation, migration, and invasion of ARPE-19 cells induced by FGF-2 (10 ng/ml) were significantly inhibited by PLB. PLB also significantly inhibits the expression of MMP-2/-9, collagen I Alpha 1 (Col1A1), collagen IV Alpha 1 (Col4A1), collagen VI Alpha 1 (Col6A1), and the phosphorylation of FGF receptor (FGFR)-1, FGFR-2, ERK, p38, JNK of FGF-2-induced ARPE-19 cells. In summary, PLB inhibits FGF-2-stimulated proliferation, migration, and invasion of ARPE-19 cells, which may take place through inhibiting the expression of MMP-2/-9, Col1A1, Col4A1, Col6A1, and the mitogen-activated protein kinase (MAPK) pathway. PLB may have a preventive effect on proliferation, migration, and invasion of FGF-2-induced ARPE-19 cells.