Kang Liu

To evaluate the effects of a multi‐strain probiotic formula, Bifidobacterium animalis subsp. lactis BLa80, Lacticaseibacillus rhamnosus LRa05, and Bifidobacterium breve BBr60, on inflammation, metabolism, and the gut microbiota in patients with type 2 diabetes mellitus (T2DM). In a randomized, double‐blind, placebo‐controlled trial, 80 adults with T2DM received either the probiotic or a placebo in addition to standard hypoglycemic therapy for 12 weeks. We assessed inflammatory cytokines, glycemic indices, serum amino acids, bile acids (BAs), short‐chain fatty acids (SCFAs), and gut microbiota composition. Compared with the placebo group, the probiotic intervention led to a significant reduction in the levels of IL‐17 and TNF‐α ( p < 0.05 ), reduced serum concentrations of threonine, isoleucine, and arginine. Additionally, the BA profile was markedly altered, revealing 16 differential metabolites that were associated with pivotal metabolic pathways. SCFA analysis showed higher isobutyric and isovaleric acid after supplementation. The probiotic group also exhibited significant reductions in total glycated hemoglobin (GHb) and fasting plasma glucose (FPG, p < 0.05). Microbiome analyses indicated decreased alpha‐diversity and distinct beta‐diversity shifts, including increased Gemmatimonadota and reduced Clostridium abundance in the probiotic group. This multi‐strain probiotic modulated inflammatory responses, metabolic profiles, including BA metabolism and SCFAs, and gut microbiota composition in T2DM, supporting its potential as an adjunct to metabolic management. Trial Registration: ClinicalTrials.gov identifier: NCT06440486

Esophageal squamous cell carcinoma (ESCC) is a highly aggressive malignancy with poor prognosis and limited therapeutic options. N6-methyladenosine (m⁶A) RNA modification plays a role in tumorigenesis, but its contributions to ESCC and the regulation of cell adhesion molecules such as Nectin-4 are not fully elucidated. In this study, we investigated the role and the regulatory mechanisms of Nectin-4 in ESCC, particularly regarding the influence of m⁶A modification and its downstream metabolic effects. Our study demonstrates that methyltransferase-like protein 3 (METTL3) enhances Nectin-4 mRNA stability and expression through m6A methylation in ESCC, as validated by actinomycin D assays, MeRIP-qPCR, and dual-luciferase reporter assays. Both METTL3 and Nectin-4 are highly expressed in ESCC tissues and promote malignant phenotypes such as proliferation, migration, and invasion. Further analysis identifies pantothenate esterase 1 (VNN1) as a downstream target of Nectin-4, mediating the oncogenic effects of the METTL3/Nectin-4 axis and promoting the biosynthesis of pantothenic acid and coenzyme A, thus driving ESCC progression. By integrating transcriptomic data, this study elucidates a key pathogenic mechanism in which the METTL3/Nectin-4/VNN1 axis regulates metabolic reprogramming to promote ESCC development. These findings provide new insights into the molecular pathology of ESCC and offer potential biomarkers and therapeutic targets for early screening, prognosis, and precision treatment.

Background: Platinum chemotherapy, particularly cisplatin, has been the standard treatment for ovarian cancer. However, the development of resistance to cisplatin is a significant challenge during treatment. Circular RNAs (circRNAs) are a class of non-coding RNAs with a circular structure and have been implicated in regulating ferroptosis and chemoresistance. Despite the increasing recognition of circRNAs in cancer progression, the role of circASH1L in ferroptosis and cisplatin resistance in ovarian cancer remains poorly understood.Methods: RNA sequencing (RNA-seq) was utilized to identify differentially expressed circRNAs in ovarian cancer cells. Cell survival and invasion were assessed using CCK-8 and transwell assays, while apoptosis, cell cycle progression, and lipid peroxidation were analyzed by flow cytometry. Levels of GSH, MDA, and iron ions were measured using appropriate kits. qRT-PCR and Western blot analyses were performed to evaluate the expression of relevant RNAs and proteins. The clinical relevance of circASH1L/miR-515-5p/CDCA7 axis in ovarian cancer patients was analyzed using public datasets. Molecular interactions were confirmed through dual-luciferase reporter assays, RNA immunoprecipitation (RIP), and co-immunoprecipitation (Co-IP). In vivo, the effects of circASH1L on ferroptosis and chemoresistance were evaluated using a xenograft mouse model.Results: circASH1L expression was downregulated upon erastin treatment and significantly upregulated in cisplatin-resistant A2780/DDP and SKOV3/DDP cells. Silencing circASH1L reversed cisplatin resistance by reducing cell viability and invasion, while promoting apoptosis and ferroptosis. Mechanistically, circASH1L was found to act as a sponge for miR-515-5p, which in turn regulates the CDCA7/RRM2 axis. Rescue experiments demonstrated that inhibiting miR-515-5p or overexpressing CDCA7 blocked the effects of circASH1L silencing. Moreover, CDCA7 could interact with RRM2 and inhibit RRM2 degradation, which contributed to reducing cell cycle arrest and ferroptosis resistance. The clinical analysis showed circASH1L, and CDCA7/RRM2 expression was positive correlated with drug resistance and worse survival rate, while miR-515-5p expression was on the contrary. In vivo, silencing circASH1L enhanced cisplatin sensitivity by inducing ferroptosis.Conclusion: Our study demonstrates that silencing circASH1L alleviates cisplatin resistance in ovarian cancer cells. The underlying mechanism involves the upregulation of miR-515-5p, which targets the CDCA7/RRM2 axis, leading to cell cycle modulation and the induction of ferroptosis. Targeting the circASH1L/miR-515-5p/CDCA7 pathway offers new insights into the relationship between ferroptosis and chemoresistance, presenting a promising strategy to overcome chemoresistance in ovarian cancer.

Background: Esophageal squamous cell carcinoma (ESCC) is a major contributor to cancer-related deaths, driven by its invasive and metastatic nature. Circular RNAs (circRNAs) are increasingly recognized as regulators of cancer progression, primarily through miRNA sponging and interactions with RNA-binding proteins. Their dysregulation has been linked to the development of in various cancers. The present study aimed to investigate the potential involvement of circSLC22A3 in the pathogenesis of ESCC. Methods: CircSLC22A3 expression in ESCC tissues and cells was analyzed using transcriptome sequencing and RT-qPCR. Its circular structure was validated through Sanger sequencing, agarose gel electrophoresis, RNase R digestion, and random priming assays. Subcellular localization was determined by nucleoplasmic separation and fluorescence in situ hybridization (FISH). Clinical correlations were assessed via tissue microarrays. Functional roles of circSLC22A3 in ESCC progression were investigated through in vitro and in vivo assays. Downstream miR-19b-3p and target gene TRAK2 were screened by bioinformatics analysis and RT-qPCR, with binding confirmed via luciferase reporter assays. RNA pulldown combined with RNA immunoprecipitation (RIP) identified IGF2BP1 as a circSLC22A3-interacting protein. RNA-seq and RT-qPCR revealed ACSBG1 as a key downstream effector. IGF2BP1-mediated m⁶A modification of ACSBG1 was mapped by MeRIP-seq and RIP, with mRNA stability assessed via Actinomycin D assay. ACSBG1 expression and biological function in ESCC were confirmed by immunohistochemistry, RT-qPCR, and functional assays. Results: Significant downregulation of circSLC22A3 was observed in both ESCC tissues and cell lines. Overexpression of circSLC22A3 significantly reduced ESCC cells’ migration and invasion capabilities. Mechanistic investigation revealed that circSLC22A3 played a pivotal role in the invasion and metastasis of esophageal cancer through distinct pathways. On one hand, circSLC22A3 functioned as a miR-19b-3p sponge to augment trafficking kinesin protein 2 (TRAK2) expression, while, on the other hand, circSLC22A3 formed a protein-RNA complex with IGF2BP1, resulting in the degradation of acyl-CoA synthetase bubblegum family member 1 (ACSBG1) mRNA through the recognition of m⁶A modification, thereby suppressing invasion and metastasis of ESCC. Conclusions: The present study identified circSLC22A3 as a new tumor suppressor that inhibited ESCC progression through both the circSLC22A3/ miR-19b-3p/ TRAK2 and circSLC22A3/ IGF2BP1/ ACSBG1 axes.

The role of basonuclin 1 (BNC1), a zinc finger protein-specific transcription factor, in gastric cancer remains unclear. In this study, BNC1 was downregulated in gastric cancer and functioned as a tumor suppressor. Through integrative analyses of transcriptome sequencing and functional assays, C-C motif chemokine ligand 20 (CCL20) was identified as a direct downstream target of BNC1. Overexpression of BNC1 inhibited the proliferation, migration, and invasion of gastric cancer cells both in vitro and in vivo. Mechanistically, BNC1 suppresses CCL20 expression by binding to its promoter, leading to reduced activation of the JAK-STAT signaling pathway and promoting apoptosis in gastric cancer cells. These findings highlight the pivotal role of BNC1 in gastric cancer progression and suggest that targeting BNC1 and its downstream pathways could serve as a potential therapeutic strategy.

Esophageal squamous cell carcinoma (ESCC) is prone to metastasis and is a leading cause of mortality. The cytoskeleton is closely related to cell morphology and movement; however, little research has been conducted on ESCC metastasis. In this study, we found that the anchoring filament protein ladinin 1 (LAD1) specifically binds to LINC01305 for co‐regulating the level of modulating cortactin proteins (CTTN) and neuronal Wiskott–Aldrich syndrome protein (N‐WASP) phosphorylation, which mediates cytoskeletal reorganization and affects the metastasis of ESCC cells. Additionally, LINC01305 and LAD1 jointly promoted the epithelial–mesenchymal transition (EMT) process by activating the phosphoinositide‐3‐kinase–protein kinase B (PI3K/AKT) signaling pathway. Moreover, LINC01305 and LAD1 were related to the late clinical stage and lymph node metastasis of ESCC. Our study demonstrated that LINC01305 and LAD1 are major determinants of ESCC dissemination and revealed a novel molecular mechanism of cytoskeletal reorganization that controls ESCC metastasis.Trial Registration: N/A.

Junctional adhesion molecule-A (JAM-A) is an adhesion molecule that exists on the surface of certain types of cells, including white blood cells, endothelial cells, and dendritic cells. In this study, the cDNA sequences of JAM-A-Fc were chemically synthesized with optimization for mammalian expression. Afterward, we analyzed JAM-A protein expression through transient transfection in HEK293 cell lines. Mice were immunized with JAM-A-Fc protein, and hybridoma was prepared by fusing myeloma cells and mouse spleen cells. Antibodies were purified from the hybridoma supernatant and four monoclonal strains were obtained and numbered 61H9, 70E5, 71A8, and 74H3 via enzyme-linked immunosorbent assay screening. Immunofluorescence staining assay showed 61H9 was the most suitable cell line for mAb production due to its fluorescence signal being the strongest. Flow cytometric analysis proved that 61H9 possessed high affinity. Moreover, antagonism of JAM-A mAb could attenuate the proliferative, migrative, and invasive abilities of ESCC cells and significantly inhibit tumor growth in mice. By examining hematoxylin-eosin staining mice tumor tissues, we found inflammatory cells infiltrated lightly in the anti-JAM-A group. The expression of BCL-2 and IκBα in the anti-JAM-A group were decreased in mice tumor tissues compared to the control group. Ultimately, a method for preparing high-yield JAM-A-Fc protein was created and a high affinity mAb against JAM-A with an antitumor effect was prepared.

Background: A clinical challenge in esophageal squamous cell carcinoma (ESCC) remains the lack of applicable plasma biomarkers for screening and diagnosis. Circular RNAs (circRNAs) hold great potential as biomarkers for cancer. The study aims to explore a circRNA as a potential plasma biomarker for screening strategies and diagnostic approaches to ESCC. Methods: Upregulated circRNAs were identified through RNA sequencing, with circCYP24A1 being identified as the target circRNA. Fluorescence in situ hybridization was employed to detect the expression of circCYP24A1 in ESCC tissue microarrays, aiming to assess the expression of circCYP24A1 in a large population and its correlation with clinical indicators. Subsequently, qRT-PCR analysis was performed on plasma samples from both ESCC patients and healthy controls to evaluate the expression levels of circCYP24A1, exploring its potential as a biomarker. Finally, the functions of circCYP24A1 were validated through CCK-8 assay, wound healing assay, trans-well assays and western blot assays. Results: CircCYP24A1 demonstrated upregulation in both plasma and tissues, exhibiting correlations with lymph node metastasis, TNM staging, and prognosis in ESCC. The circCYP24A1 achieved a perfect area under the curve of 0.94 for the diagnosis of ESCC, and an area under the curve of 0.76 for the prediction of lymph node metastasis. Furthermore, functional loss assays revealed that circCYP24A1 effectively promotes the epithelial-mesenchymal transition and tumor metastasis in vitro. Conclusions: CircCYP24A1 emerges as a potential plasma diagnostic biomarker and a predictive factor for LNM for ESCC.

EsophageaL squamous cell carcinoma (ESCC) is one of the most common and lethal tumors, however, its underlying molecular mechanisms are not completely understood and new therapeutic targets are needed. Here, we found that the transcription factor basonuclin 1 (BNC1) was significantly upregulated and closely related to the differentiation and metastasis of ESCC. Furthermore, BNC1, LINC01305, and G‐protein pathway suppressor 1 (GPS1) had significant oncogenic roles in ESCC. In addition, in vivo experiments showed that knockdown of BNC1 indeed significantly inhibited the proliferation and metastasis of ESCC. We also revealed the molecular mechanism by which LINC01305 recruits BNC1 to the promoter of GPS1, and then GPS1 could mediate the JNK signaling pathway to promote the proliferation and metastases of ESCC. Taken together, we discovered the novel molecular mechanism by which LINC01305/BNC1 upregulates GPS1 expression to promote the development of ESCC, providing a new therapeutic target for ESCC.