Hepatocellular carcinoma (HCC) is a common malignant tumor worldwide. The specific role of Sirtuin3 (SIRT3), a member of the Sirtuin family, in HCC, particularly regarding drug sensitivity, has not been fully elucidated. This study aimed to elucidate the mechanism by which SIRT3 modulates drug sensitivity in HCC. Expression prediction of target genes, survival analysis, and candidate protein screening were conducted using bioinformatics databases. After transfection of different plasmids in HCC cells, the expression of target genes was detected using RT-qPCR and Western blot. CCK-8 assay was used to assess the effect of SIRT3 overexpression on the proliferation ability of HCC cells. Additionally, changes in ferroptosis levels in HCC cells were reflected by detecting ferroptosis-related indicators (including ROS, MDA, GSH, and Fe2+ levels). SIRT3 was downregulated in HCC and its overexpression promoted sensitivity of HCC cells to oxaliplatin. Mechanistically, SIRT3 induced deacetylation modification and ubiquitination modification of GRP78, leading to a decrease in its stability and subsequent degradation of GRP78 protein. This process limited the correction of misfolded proteins during endoplasmic reticulum stress (ERS). The inhibition of the downstream transcription factor of ER stress ATF4 activation leads to the suppression of the anti-ferroptosis gene NRF2 transcription, thereby promoting ferroptosis in HCC cells and enhancing their sensitivity to oxaliplatin. In summary, SIRT3 enhances the sensitivity of HCC cells to oxaliplatin by promoting ferroptosis through the inhibition of GRP78-mediated ERS and regulation of the ATF4/NRF2 axis.

Liver metastasis is a frequent complication in colorectal cancer (CRC), significantly impacting patient prognosis. This study aims to develop a machine learning-based prediction model for metachronous liver metastasis (MLM) in CRC patients, facilitating early diagnosis and intervention to potentially improve treatment outcomes and survival rates. A retrospective analysis was conducted on 620 consecutive patients who underwent radical colorectal cancer resection at the First People’s Hospital of Changzhou during the study period and met the predefined inclusion and exclusion criteria. MLM status was determined according to postoperative follow-up outcomes rather than used as a sampling criterion. Among the final eligible primary cohort, 373 patients had no observed liver metastasis during follow-up, whereas 247 patients were diagnosed with metachronous liver metastasis more than 6 months after radical CRC resection. Patients were split into training (non-MLM = 258, MLM = 176) and internal validation (non-MLM = 258, MLM = 71) cohorts, with an external cohort of 52 non-MLM and 30 MLM patients from the Seventh People’s Hospital of Changzhou. Missing values were imputed using KNN. Based on the features selected by Logistic Regression (LR) and LASSO, five machine learning models (LR, RF, LightGBM, XGBoost, and SVM) were developed. Model performance was comprehensively evaluated using AUROC, DCA, accuracy, sensitivity, specificity, and F1 score, with SHAP illustrating feature influence. The best model was validated internally and externally. Eight independent risk factors (Age, Tumor Embolus, Size, T stage, N stage, Tumor Differentiation, RDW, AST) were included as features in the model. The LR model demonstrated the best performance, with SHAP indicating T stage as the most influential factor. A dynamic online nomogram was constructed to visualize the LR model. ROC analysis showed good discriminative performance of the final LR model, and calibration analysis suggested acceptable agreement between predicted and observed MLM risk. DCA and CIC analyses suggested potential clinical net benefit and clinical impact within the prespecified clinically relevant threshold probability range. In this study, LR model was selected as the final model for predicting the risk of metachronous liver metastasis after radical CRC resection. The dynamic nomogram provides an interpretable and accessible tool for visualizing individualized risk estimates. Given the retrospective design and current validation limitations, the model should be regarded as a supplementary tool to support postoperative risk stratification and follow-up planning rather than as definitive evidence for clinical decision-making. Further prospective and multicenter validation is warranted before routine clinical implementation.

Robust liver regeneration counteracts and facilitates recovery from liver injuries. The underlying epigenetic mechanisms, however, are not fully understood. Here we investigated the role of suppressor of variegation 3-9 homolog 1 (Suv39h1), a histone H3K9 methyltransferase, in liver regeneration. Suv39h1 expression was repressed by DNMT1 during liver regeneration. Systemic or hepatocyte-specific deletion of Suv39h1 in mice enhanced liver regeneration and post-surgery survival following partial hepatectomy. RNA sequencing revealed high-mobility group protein B2 (HMGB2) as a target for Suv39h1. Suv39h1 downregulation in proliferating hepatocytes allowed E2F1 to activate HMGB2 transcription. Consistently, HMGB2 knockdown attenuated proliferation of hepatocytes in response to HGF treatment and suppressed liver regeneration in mice. Integrated transcriptomic analysis indicated that HMGB2 may contribute to proliferation of hepatocytes by regulating a panel of proregenerative genes. Importantly, Suv39h1 inhibition by chaetocin boosted liver regeneration in mice. Finally, a significant correlation between Suv39h1, HMGB2 and proliferative markers was identified in patients with acute liver failure. In conclusion, our data uncover an unrecognized role for Suv39h1 in liver regeneration. Therefore, targeting Suv39h1 may be considered as a viable strategy to boost liver regeneration after injury.

BACKGROUND:Radiopharmaceutical therapy uses drugs with radioactive isotopes for targeted cancer treatment. However, its effectiveness in gastric cancer is often limited by poor tumor penetration, heterogeneous intratumoral distribution, and off-target radiation exposure. Ultrasound-guided microbubble-assisted delivery offers a noninvasive means to increase vascular permeability and enhance tumor-specific radiopharmaceutical uptake. METHODS:This study developed and evaluated an ultrasound-guided microbubble delivery system to enhance intratumoral radiopharmaceutical accumulation, therapeutic efficacy, and safety in gastric cancer. Ligand-functionalized, radiopharmaceutical-loaded microbubbles were administered intravenously in an orthotopic gastric tumor model. Tumors were localized by real-time ultrasound imaging and then exposed to focused ultrasound to induce controlled microbubble cavitation. Biodistribution was analyzed using positron emission tomography imaging. Tumor growth was monitored by serial ultrasound, and therapeutic response was assessed with histopathology and Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays. Radiation safety was determined by organ-specific biodosimetry. RESULTS:Microbubble activation significantly increased intratumoral uptake to 8.4-1.2%ID/g, compared with 3.0 - 0.6%ID/g for conventional delivery (p < 0.001). Tumor-to background ratios improved approximately 2.6-fold, and tumor volumes decreased by 58% relative to controls. Apoptotic indices reached 41.8 - 6.3% in treated tumors versus < 15% in controls; off-target organ uptake remained < 2.0% ID/g. CONCLUSIONS:Ultrasound-guided, microbubble-assisted radiopharmaceutical administration increased tumor-specific absorption, slowed tumor development, and decreased systemic exposure, indicating a promising treatment strategy for gastric cancer.

During chronic liver injury, danger signals released by damaged hepatocytes are known to sequentially promote the polarization of monocyte-derived macrophages (Mo-Macs) toward a pro-fibrotic phenotype and activate hepatic stellate cells (HSCs), ultimately leading to hepatic fibrosis (HF). Intracellular endoplasmic reticulum (ER) stress, tightly associated with dysregulated calcium ion (Ca2+) flux and excessive reactive oxygen species (ROS) production, is recognized as a key driver of this pro-fibrotic polarization process. Recently, metal-polyphenolic networks (MPNs) have garnered significant attention as a versatile class of nanomedicines for treating various diseases, with their potential for tailored design and functional modification, as well as the stimuli-responsive delivery of natural polyphenols and metal ions. In this study, exploiting the ability of magnesium ions (Mg2+) to suppress ER Ca2+ release and tannic acid (TA) to scavenge ROS, we fabricated Mg-TA MPN-based nanomedicines aimed at restoring ER homeostasis for the treatment of HF. The resulting PEGylated formulation, Mg-TA-PEG (MTP), exhibited excellent biocompatibility, pH-responsive dissociation, and efficient hepatic accumulation. Intravenous MTP administration significantly attenuated carbon tetrachloride induced HF in mice, as evidenced by reduced collagen deposition, normalized liver architecture, and improved metabolic function. Single-nucleus RNA sequencing analysis revealed that MTP inhibits the induction of Mo-Macs by damaged hepatocytes, alleviates ER stress in Mo-Macs, and shifts their polarization from a fibrosis-promoting phenotype to immuno-suppressing and regeneration-promoting phenotypes. This reprogramming further modulates paracrine signaling from Mo-Macs to HSCs, reverting HSCs from an activated to a quiescent state. Moreover, in vitro cell coculture experiments confirmed that MTP primarily targets Mo-Macs rather than directly inhibiting HSC activation, underscoring the central role of Mo-Macs in MTP mediated fibrosis resolution. Together, these findings elucidate the anti-fibrotic mechanism of MTP nanomedicines and highlight a promising strategy for reprogramming Mo-Mac phenotypes to treat HF.

Inflammatory disease is sustained not only by immune activation but also by the failure of activated cells to terminate effector programs and return tissues to homeostasis. This review evaluates N6-methyladenosine (m6A) as a post-transcriptional regulator of that persistence. Rather than cataloguing diseases or classifying individual m6A regulators as pro- or anti-inflammatory, we organize the evidence around a causal chain linking a regulator and target RNA to a site- or reader-dependent change in RNA fate, an immune-cell phenotype, and a disease outcome. Mechanistically developed evidence is concentrated in macrophages and T cells, including STAT1 stability and decay, SOCS turnover, SLC37A2 translation, and lineage-specific T-cell programs. Mechanisms in other innate and adaptive immune populations are emerging, but cell-subset resolution and disease-stage validation remain uneven. Across inflammatory signaling, immunometabolic, and cell-death pathways, the same m6A regulator can produce opposing outcomes through different target transcripts or readers. We therefore distinguish mechanistically resolved, functional/intermediate, and associative evidence while highlighting the limitations of global m6A assays, bulk-tissue profiling, and pathway-level inference. The translational value of m6A is likely to depend on target-RNA selection, cell-specific delivery, disease-stage timing, and preservation of protective immunity and tissue repair. An evidence-aware, transcript-centered framework may help move m6A research from descriptive association toward causal immune-state biology and clinically testable interventions.

Microbe-derived particles (MDPs), which include extracellular vesicles, outer membrane vesicles, inclusion bodies, polysaccharide particles, and virus-like particles, represent a rapidly expanding category of bioinspired nanomaterials. With their natural origin, intrinsic biocompatibility, and highly programmable functionality, MDPs serve as a versatile bridge between living microbial systems and engineered nanomaterials. This review systematically outlines the diverse classes, biosynthetic pathways, physicochemical properties, and biomedical applications of MDPs. We summarize recent progress in their use for drug delivery, vaccine design, immune modulation, and diagnostic imaging, emphasizing how bioengineering approaches enable precise control over their composition and targeting capabilities. Additionally, we examine advancing methodologies for scalable manufacturing and accurate characterization-such as omics-based profiling and advanced imaging techniques. Finally, we discuss current challenges and future prospects for integrating synthetic biology with materials science to develop next-generation intelligent biological platforms based on MDPs. This comprehensive overview aims to foster the rational design and translational deployment of MDP-based systems across biomedical, biotechnological, and materials science fields.

With the advancement of hydrogel technology, increasing attention has been drawn to hydrogel microspheres (HMs) due to their versatile biomedical applications. HMs play pivotal roles in biomedical applications, such as drug delivery, cell culture, regenerative medicine, wound healing, and tumor immunity. Composed of diverse biobased materials and fabricated through various preparation methods, HMs offer unique structural and functional advantages. This review focuses on the latest findings to provide a more comprehensive understanding of HMs for biomedical applications. Their therapeutic potential across multiple disease contexts is highlighted, and emerging trends and challenges are discussed. By consolidating current knowledge, this work aims to inspire further research and accelerate the clinical translation of HMs.

Background:After distal pancreatectomy (DP), clinically relevant postoperative pancreatic fistula (CR-POPF) is a critical complication that adversely affects the prognosis of patients. The present study identified the risk factors for CR-POPF occurrences, as well as developing a nomogram to predict their risk after DP. Methods:We retrospectively examined 300 medical records, obtaining the patients' preoperative clinical baseline data, laboratory indicators, preoperative computed tomography (CT) data, and intraoperative clinical information. We determined CR-POPF independent risk factors using univariate as well as multivariate logistic regression analyses. We created a risk nomogram based on these variables and used the bootstrap method for internal validation. Area under the curve (AUC) assessed the nomogram's predictive power. The nomogram's clinical value and viability were evaluated using decision curve analysis (DCA) as well as clinical impact curve (CIC). Results:CR-POPF developed in 84 of the 300 patients (28.0% incidence). CR-POPF was found to be independently risked by operation time (P=0.002), preoperative C-reactive protein (CRP) levels (P<0.001), CT (pancreas)-to-CT (psoas major) ratio (P<0.001), and pancreatic thickness (PT) at transection site (P<0.001). The nomogram's AUC was 0.901, which, along with the DCA, highlighted the nomogram's excellent performance, surpassing those of four alternative CR-POPF prediction models. The nomogram has an immense net clinical advantage, according to the CIC. Conclusions:The developed nomogram can be useful in identifying high-risk patients and formulating individualized perioperative plans to prevent the risk of CR-POPF formation in patients undergoing DP.

Copper (Cu)-dependent aggregation of lipoylated proteins in mitochondria triggers cuproptosis, a newly discovered modality of regulated cell death marked by impaired mitochondrial respiration. During the progression of hepatic fibrosis (HF), hepatocytes, responsible for Cu storage and metabolism, exhibit vigorous mitochondrial activity and are supposed to be cuproptosis-susceptible. However, the cellular landscape of cuproptosis in fibrotic livers remains unclear, and the therapeutic potential of cuproptosis suppression in resolving HF has yet to be explored. In this study, single-cell RNA sequencing data from mouse models and clinical samples from HF patients show that hepatocytes are the dominant cell population attacked by cuproptosis. These cuproptotic hepatocytes release pro-fibrotic signals that activate hepatic stellate cells (HSCs), inducing the formation of fibrotic scar. Tannic acid (TA), a polyphenolic-molecule cuproptosis inhibitor, can combat the changes in cuproptotic hepatocytes stimulated by elesclomol plus Cu supplement in vitro, including intracellular Cu (I) concentration, mitochondrial membrane potential, and expression pattern of cuproptosis regulators. In addition, the production of pro-fibrotic signals is inhibited by TA, and subsequently the activation of HSCs. Finally, TA is demonstrated to prevent the progression of HF in mouse models, as evidenced by the improvement of liver architecture and function. Proteomic analysis reveals that the disturbed lipid metabolism and cuproptosis regulator expression in fibrotic livers are both rescued by TA. Collectively, these findings highlight hepatocyte cuproptosis as a key driver of the fibrotic process, and its suppression holds great therapeutic promise for treating HF.

Ferritinophagy, the selective autophagic degradation of ferritin to release iron, is emerging as a critical regulator of iron homeostasis and a key player in the pathogenesis of various liver diseases. This review comprehensively examines the mechanisms, regulation, and multifaceted roles of ferritinophagy in liver health and disease. Ferritinophagy is intricately regulated by several factors, including Nuclear Receptor Coactivator 4 (NCOA4), Iron regulatory proteins and signaling pathways such as mTOR and AMPK. These regulatory mechanisms ensure proper iron utilization and prevent iron overload, which can induce oxidative stress and ferroptosis. In liver diseases, ferritinophagy exhibits dual roles. In liver fibrosis, promoting ferritinophagy in hepatic stellate cells (HSCs) can induce cell senescence and reduce fibrosis progression. However, in non-alcoholic fatty liver disease (NAFLD), chronic ferritinophagy may exacerbate liver injury through iron overload and oxidative stress. In hepatocellular carcinoma (HCC), ferritinophagy can be harnessed as a novel therapeutic strategy by inducing ferroptosis in cancer cells. Additionally, ferritinophagy is implicated in drug-induced liver injury and sepsis-associated liver damage, highlighting its broad impact on liver pathology. This review also explores the crosstalk between ferritinophagy and other selective autophagy pathways, such as mitophagy and lipophagy, which collectively influence cellular homeostasis and disease progression. Understanding these interactions is essential for developing comprehensive therapeutic strategies targeting multiple autophagy pathways. In summary, ferritinophagy is a complex and dynamic process with significant implications for liver diseases. This review provides an in-depth analysis of ferritinophagy’s regulatory mechanisms and its potential as a therapeutic target, emphasizing the need for further research to elucidate its role in liver health and disease.

Background:Placement of a support tube during pancreaticojejunal anastomosis has been shown to reduce the incidence of postoperative pancreatic fistula in patients with a non-dilated pancreatic duct. However, increasing reports of long-term complications, such as the presence of an indwelling support tube in the anastomosis, have raised concerns. Despite this, short-term complications, such as gastrointestinal perforation in the absence of tube displacement, remain relatively rare. Case Description:This report describes a 59-year-old female patient who underwent radical pancreaticoduodenectomy for pancreatic ductal adenocarcinoma (T2N0M0). On the 8th postoperative day, she developed back pain, followed by a sudden increase in body temperature, elevated inflammatory markers, and a significant increase in amylase levels in the drainage fluid. Enhanced abdominal computed tomography revealed that one side of the pancreatic duct support tube had penetrated the abdominal cavity, resulting in posterior peritoneal effusion. Emergency removal of the support tube and repair of the intestinal perforation were performed. The patient had an uneventful recovery after surgery and was discharged on day 20 following the second operation. Conclusions:In cases of postoperative abdominal or low back pain accompanied by signs of infection, complications related to the support tube should be considered. Further studies are needed to evaluate the necessity of placing a pancreatic duct support tube, as well as the timing for its removal after the completion of the anastomosis.

Background The anatomical relationship between the pancreatic tail and splenic vessels influences both pancreatic resection completeness and spleen preservation success in robotic spleen-preserving left-sided pancreatectomy (RSPLP). However, surgical strategies for managing the pancreatic tail during RSPLP remain unreported Methods Clinical data from 46 consecutive patients undergoing robot-assisted left-sided pancreatectomy with intended spleen preservation were analyzed. Pancreatic tails were classified into four anatomical types (I-IV) based on their relationship with splenic vessels or tumors. Results The RLP group (spleen preservation failure) exhibited a higher proportion of type IV anatomy than the RSPLP group (spleen preservation success) (76.92% vs. 18.18%, P<0.05). Type III/IV anatomy was associated with significantly lower spleen preservation rates (52.17% vs. 91.30%, P=0.003), larger tumor size [4.0 (3.5-5.8) vs. 2.8 (2.2-4.6) cm, P=0.026], greater intraoperative blood loss [134 (58-295) vs. 85 (45-180) mL, P=0.017], and longer operative time (257.3±62.29 vs. 220.78±53.05 min, P=0.038) compared to type I/II. Conclusion We proposed the preoperative classification of pancreatic tail type and the “pendulum separation” technique, and found that it was challenging to preserve the spleen in type III and IV pancreatic tails due to their complex anatomical relationship with the splenic vessel and tumor.

e16440 Background: This study aimed to investigate overall survival (OS) in patients with surgically resected pancreatic cancer and examined the distribution of intratumoral microbiomes across distinct sites within pancreatic tumors. Methods: Patients were selected from the Surveillance, Epidemiology, and End Results (SEER) registry (2000–2021 dataset). Kaplan-Meier survival analysis and multivariable Cox proportional hazards models were used to assess overall and site-specific survival outcomes. A retrospective analysis was initiated to verify prognostic factors related with survival in surgically resected pancreatic cancer patients. To compare differential profiles of intratumoral microbiota, 16S rRNA gene sequencing was conducted on tissue samples from 7 patients with pancreatic head cancer (PHC) and 7 with pancreatic body/tail cancer (PBTC). Correlations between microbiota and prognosis, as well as associated gene expression changes, were analyzed using data from The Cancer Genome Atlas (TCGA). The effect of microbiota on tumor metastasis was confirmed by a pulmonary metastasis model of mouse pancreatic cancer. Results: A total of 78,548 patients were identified. The OS of surgically resected patients with PBTC was significantly better than that of patients with PHC. Multivariate analysis showed that male, older age, pancreatic head location, M1 and stage IIB/III/IV were independent predictors of poorer survival. Retrospective analysis involved 162 patients of our center verified that OS of PBTC was significantly longer than PHC. Microbiota was detected in tumor tissue of pancreatic cancer patients. The microbial species diversity in PHC tissues was significantly higher than in PBTC tissues, with microbial compositions across six phylogenetic levels highlighting specific differences between PHC and PBTC patients. At the genus level, 17 bacterial genera were identified, among which Sphaerotilus and Dyella , significantly enriched in PBTC patients, were associated with improved survival, as indicated by TCGA survival data. Absence of these bacteria might contribute to immune deficiency, widespread activation of cancer-related pathways, and metabolic reprogramming. In vivo experiment proved that Sphaerotilus and Dyella could inhibit pulmonary metastasis of pancreatic cancer cells. Conclusions: Our study demonstrated that patients with pancreatic body/tail tumors had a notably longer median survival than those with tumors in the head region following surgical resection. Enrichment of Sphaerotilus and Dyella in PBTC was significantly associated with improved survival in pancreatic cancer patients. These findings suggest that intratumoral microbiota may play a role by modulating signaling-related genes and regulating immune status, highlighting its potential as a therapeutic target and prognostic marker of pancreatic cancer patients.

BACKGROUND:Radiation-induced liver injury (RILI) poses a significant challenge in abdomino-pelvic tumor radiotherapy, adversely impacting normal liver tissues. This study explores the role of Tmprss6, a gene encoding Matriptase2, in acute RILI development and its impact on hepatocyte apoptosis. METHODS:A RILI model was established using 30 Gy total liver irradiation in C57BL/6J mice. Expression of Tmprss6 and hepcidin post radiation was detected by immunohistochemistry staining, Western blot and quantitative real-time PCR. Tmprss6 knockout mice were established to demonstrate the contribution of Tmprss6 to RILI. The molecular response within the liver after Tmprss6 knockdown was investigated using Transcriptomics sequencing. RESULTS:Disruptions in the Matriptase2-hepcidin axis emerge post-total liver radiation, marked by decreased Matriptase2 levels and heightened hepcidin levels. Tmprss6 knockout exacerbates RILI, evident in reduced mouse survival, increased liver damage, and elevated serum levels of aminotransferases. Additionally, Tmprss6 deletion increases inflammatory cytokines and malondialdehyde level while diminishing liver antioxidant capacity. Gene expression profiling reveals shifts in inflammation, apoptosis, and p53 signaling pathways upon Tmprss6 deletion. In vitro experiments, utilizing the AKT inhibitor AZD5363, demonstrate its effectiveness in reversing impediments caused by Tmprss6 silencing post-radiation. AZD5363 effectively restores suppressed cell proliferation, mitigates heightened cell apoptosis, and counters the elevated p53 expression induced by Tmprss6 depletion. This underscores the partial mediation of Tmprss6's protective role through the PI3K-AKT signaling. CONCLUSIONS:This study unveils intricate mechanistic pathways activated by Tmprss6 silencing, amplifying p53 expression, facilitating hepatocyte apoptosis, and accelerating RILI progression, which providing nuanced insights into the multifaceted involvement of Tmprss6 in RILI.

The treatment outcomes of traditional patches for chronic soft tissue defects (CSTDs) are unsatisfactory in clinical, owing to the lack of intrinsic bioactivities to orchestrate the intricate regenerative process. To tackle this deficiency, nature-derived microneedles (NMs) composed of silk methacrylate and snail mucus are developed in this study. The resultant NMs have excellent mechanical strength and biological adhesiveness, ensuring suture-free but reliable fixation on implanted site. To enhance the intrinsic bioactivities, metal-polyphenolic networks (MPNs) coordinated from copper (Cu) and curcumin (Cur) are designed and encapsulated into NMs. Cu-Cur MPNs harness the anti-oxidative and anti-inflammatory properties of Cur with the pro-angiogenic properties of Cu, targeting different negative aspects in CSTDs repair. Furthermore, the pH-responsive disassembly of Cu-Cur MPNs can respond to the acidic microenvironment, allowing for burst-free and on-demand drug delivery. Both in-vitro and in-vivo experiments demonstrate that NMs with Cu-Cur MPNs encapsulation (Cu-Cur-NMs) can restore redox homeostasis, reduce inflammatory response, and promote blood vessel formation, thus remodeling the regenerative microenvironment to greatly improve the repair quality of CSTDs. Therefore, the combined advantages of microneedles-based patch system and MPNs-based nanotherapeutic agent are explored for the first time, and our proposed Cu-Cur-NMs represent a multifunctional and promising device for CSTDs repair.

622 Background: Cholangiocarcinoma (CCA) is a highly malignant biliary tumor characterized by frequent perineural invasion (PNI), which is associated with a poor prognosis; however, the underlying mechanisms remain unclear. G-protein-coupled receptor 120 (GPR120) has been implicated in the development of various tumors, while nerve growth factor (NGF) has a strong correlation with PNI. This study aims to elucidate the role of GPR120 in the production of NGF and to investigate the mechanisms by which PNI is induced in CCA. Methods: We conducted a retrospective review of medical records for 386 patients with cholangiocarcinoma (CCA), including both extrahepatic (ECC) and intrahepatic (ICC) types, who underwent curative resection at the Department of General Surgery, First Affiliated Hospital of Soochow University, between January 2015 and December 2021. NGF and GPR120 expression were assessed in 60 paraffin-embedded archived CCA tissue samples via immunohistochemical staining (IHC), with an additional 60 normal bile duct samples serving as controls. All tissues were sourced from the aforementioned 386 patients, none of whom received preoperative treatment. Tumors were staged according to AJCC classification. Western blot analysis was employed to evaluate NGF and GPR120 expression in CCA cell lines (QBC 939, RBE, HUCCT-1, and HCCC-9810). To assess the invasion and migration capabilities of CCA cell lines, QBC 939 and HCCC-9810 were treated with TUG-891 (a GPR120 agonist) and AH7614 (a GPR120 inhibitor). Results: Both GPR120 and NGF were found to be upregulated in CCA tissues, correlating with aggressive clinicopathological features. IHC staining revealed cytoplasmic localization of GPR120 and NGF, with significantly higher expression in cancerous tissues compared to adjacent noncancerous samples. Among the 386 patients, PNI was observed in 339 (87.8%). PNI correlated with tumor diameter, CA 19-9 levels, TNM stage, preoperative bilirubin levels, tumor differentiation, and preoperative drainage. Notably, GPR120 expression increased in poorly differentiated tumors and advanced clinical stages, indicating a clinical association with CCA progression. In vitro studies demonstrated that GPR120 promoted invasion and migration in the QBC 939 and HCCC-9810 cell lines. Western blot analysis confirmed elevated GPR120 levels in these lines, which also exhibited higher NGF expression. The GPR120 inhibitor AH7614 significantly reduced invasion in QBC 939 and HCCC-9810 cells, while the GPR120 agonist TUG-891 showed no effect on any of the CCA cell lines tested. Conclusions: GPR120 and NGF are implicated in perineural invasion (PNI) in CCA, with a notable correlation between GPR120 and NGF in the context of PNI occurrence.

Objective Hepatocellular carcinoma (HCC) is a lethal malignancy in the world. LncRNA THUMPD3-AS1 is implicated in tumorigenesis and progression in various tumors. Therefore, this study was applied to investigate the action of THUMPD3-AS1 in HCC by regulating microRNA (miR)-4465 and KPNA2. Methods The clinical specimens of HCC were collected to determine THUMPD3-AS1, KPNA2, miR-4465, E-cadherin, Vimentin, N-cadherin, ZEB1 and SNAIL levels. HCC cells were screened and transfected with sh-THUMPD3-AS1 or miR-4465 mimic to explore their roles in HCC cell phenotype and epithelial-mesenchymal transition (EMT)-related factors. The involvement of miR-4465 in THUMPD3-AS1-mediated HCC was proved. The relationship of THUMPD3-AS1, KPNA2 and miR-4465 was verified. Results Overexpressed THUMPD3-AS1 and KPNA2 and reduced miR-4465 were present in HCC clinical tissues. THUMPD3-AS1 bound to miR-4465 to target KPNA2. Silencing of THUMPD3-AS1 or restoration of miR-4465 repressed HCC cell phenotypes and EMT in vitro. Inhibition of miR-4465 mitigated the role of silenced THUMPD3-AS1 in HCC. Conclusion This study stresses that THUMPD3-AS1 induces EMT in HCC cells and ultimately promotes HCC cell growth and migration by competitively inhibiting miR-4465 expression and thus upregulating KPNA2.