4009 Background: The addition of immune checkpoint inhibitors (ICIs) to chemotherapy (chemo) in the neoadjuvant/adjuvant setting for gastric cancer (GC) has yielded mixed efficacy. Target population and treatment regimen remain to be optimized. This study compared the efficacy of serplulimab (anti-PD-1 antibody) plus chemo versus placebo plus chemo as neoadjuvant therapy and serplulimab versus chemo as adjuvant therapy for resectable GC. Methods: Patients with PD-L1 positive (PD-L1 CPS ≥ 5), histologically confirmed, untreated, resectable gastric or gastroesophageal junction (GEJ) adenocarcinoma were randomized 1:1 to receive 3 cycles of neoadjuvant intravenous (iv) serplulimab (4.5 mg/kg) or placebo, plus chemo (iv oxaliplatin, 130 mg/m 2 and oral S-1, 40-60 mg based on body surface area), followed by post-surgery adjuvant serplulimab monotherapy (up to 17 cycles) or adjuvant chemo (5 cycles), Q3W. The primary endpoint was investigator (INV)-assessed event-free survival (EFS). Efficacy superiority was first evaluated and established in patients with PD-L1 CPS ≥ 10 followed by that in the PD-L1 CPS ≥ 5 population. Results: Between November 26, 2019 and April 19, 2024, 588 patients were enrolled across 57 sites, with 292 randomized to the serplulimab group and 296 to the placebo group. As of the data cutoff date of August 19, 2025 with a median follow-up duration of 35.9 months, INV-assessed median EFS was significantly longer in the serplulimab group (n = 193) than in the placebo group (n = 217) in the PD-L1 CPS ≥ 10 population (not reached [NR], 95% CI 43.0–not estimable [NE] vs. 42.0 months, 95% CI 20.5–NE; HR 0.65, 95% CI 0.47–0.90; p = 0.0082), and in the PD-L1 CPS ≥ 5 population (NR, 95% CI 37.9–NE vs. 35.9 months, 95% CI 21.3–52.0; HR 0.73, 95% CI 0.56–0.94; p = 0.0152), meeting the protocol-specified superior criteria. Blinded independent central review-assessed median EFS was consistent with that of the investigator. Pathological complete response rate was higher in the serplulimab group than in the placebo group (21.6% vs 6.4%; odds ratio 3.95). Median OS was immature in both groups. Grade ≥ 3 treatment-related adverse events (TRAEs) were reported in 136 (46.6%) and 172 (58.5%) patients in the serplulimab and placebo groups, respectively. Discontinuation of any component of the study regimen due to TRAEs occurred in 19 (6.5%) and 31 (10.5%) patients in the respective groups. Conclusions: Neoadjuvant serplulimab plus chemo, followed by adjuvant serplulimab monotherapy, significantly prolonged EFS in patients with PD-L1-positive, resectable GC or GEJ adenocarcinoma. Improvements in other efficacy endpoints were also observed along with a favorable safety profile compared to neoadjuvant/adjuvant chemo. This world-first chemotherapy-free regimen (adjuvant) represents a promising treatment option for this indication. Clinical trial information: NCT04139135 .

Background: Triple-negative breast cancer (TNBC) features a unique molecular phenotype and high heterogeneity, posing significant challenges in clinical management and prognosis. Cuproptosis is a newly identified form of copper ion-dependent programmed cell death, but its role and molecular landscape in TNBC remain poorly understood. Methods: This study integrated bulk RNA sequencing and single-cell RNA sequencing data to systematically analyze the expression profiles and functions of cuproptosis-related genes (CRGs) in TNBC. Results: We first identified CRGs that were widely dysregulated in TNBC and classified patients into two cuproptosis regulatory patterns subtypes with distinct prognostic outcomes based on their expression profiles. Cluster 1 exhibited poorer overall survival, immunosuppression microenvironment characteristics, and active invasion-related pathways. Furthermore, we constructed and validated a prognostic risk signatures based on four key CRGs (SLC31A1, DBT, MTF1, DLST). This signature effectively distinguished high-risk from low-risk patients in both the training and independent validation cohorts, with the risk score serving as an independent prognostic factor. Combined with clinical characteristics, we developed a nomogram predicting TNBC survival rates, demonstrating excellent predictive accuracy. Tumor microenvironment analysis revealed that the high-risk group exhibited a unique immune cell infiltration profile and higher immune checkpoint molecule expression, suggesting potential increased sensitivity to immunotherapy. Drug sensitivity prediction revealed that high-risk patients might exhibit heightened responsiveness to multiple multiple chemotherapy and targeted therapy drugs. IHC experiments further validated the expression of key CRGs in TNBC tissues. Besides, through single-cell transcriptomic analysis, we identified markedly elevated copper death activity in TNBC tissues and pinpointed T cells as the key effector cell population. Cell communication analysis further uncovered an abnormally active communication network involving T cells, plasma cells, and fibroblasts within TNBC. Conclusion: This study comprehensively revealed cuproptosis heterogeneity in TNBC and the interaction with the tumor immune microenvironment, providing new insights and potential targets for TNBC prognosis prediction and personalized therapy.

The development of novel therapeutic strategies for advanced and metastatic hepatocellular carcinoma (HCC) remains an urgent clinical need. Despite suboptimal efficacy, the breakthrough of tyrosine kinase inhibitors in HCC treatment therapy underscores the advantage of targeted therapy. Therefore, innovative targeted therapies are urgently needed to enhance treatment efficacy, decrease recurrence rates, and improve patient survival outcomes. The forkhead box M1 (FOXM1) transcription factor serves as a master regulator of oncogenic signaling networks that drive cancer progression. Our study identified budding uninhibited by benzimidazoles 1 (BUB1) as a crucial downstream effector of FOXM1, with demonstrated direct protein-protein interaction. Moreover, FOXM1 directly bound to the GTAAACC motif at the -293 bp region of the BUB1 promoter and activated its transcription, thereby driving HCC cell proliferation. Mechanism studies have shown that the FOXM1/BUB1 axis regulated multiple oncogenic processes in HCC, including cell proliferation, DNA repair, G2/M cell cycle transition, stemness, invasion, and migration. Knockdown of BUB1 significantly sensitized HCC cells and xenograft tumors to the FOXM1 inhibitor FDI-6. Furthermore, combined pharmacological inhibition of FOXM1 (FDI-6, RCM-1, thiostrepton) and BUB1 (BAY-1816032) synergistically inhibited the proliferation of HCC cells and xenograft tumors. These findings establish FOXM1-mediated BUB1 upregulation as a key driver of HCC malignancy. Targeting the FOXM1/BUB1 axis represents a promising therapeutic strategy for the treatment of advanced and metastatic HCC, offering new opportunities for HCC therapy.

BACKGROUND:Perioperative chemo-immunotherapy shows variable outcomes in resectable gastric or gastro-oesophageal junction adenocarcinoma. We evaluated the efficacy and safety of neoadjuvant serplulimab with S-1 plus oxaliplatin (SOX) chemotherapy followed by adjuvant serplulimab versus neoadjuvant placebo plus SOX followed by adjuvant SOX in patients with PD-L1-positive, locally advanced, resectable gastric or gastro-oesophageal junction adenocarcinoma. METHODS:In this randomised, double-blind, multicentre phase 3 ASTRUM-006 study, patients were screened at 75 hospitals in China and Thailand. Eligible patients aged 18-70 years with PD-L1 combined positive score (CPS) ≥5, resectable gastric or gastro-oesophageal junction adenocarcinoma were randomly assigned 1:1 to neoadjuvant serplulimab or placebo (intravenous; 4·5 mg/kg) plus SOX chemotherapy (intravenous oxaliplatin 130 mg/m2 on day 1, and oral S1 40-60 mg twice daily on days 1-14) for three cycles (cycle length 21 days), followed by adjuvant serplulimab (up to 17 cycles; serplulimab group) or SOX (five cycles; placebo group). The primary endpoint was investigator-assessed event-free survival (defined as the time from randomisation to the occurrence of progressive disease or local or distant recurrence, other new malignancies, or death), with efficacy first evaluated in PD-L1 CPS ≥10, then in the intention-to-treat (CPS ≥5) population. This study is registered with ClinicalTrials.gov, NCT04139135, and is ongoing. FINDINGS:Between Nov 26, 2019, and April 19, 2024, 1646 patients were screened, of whom 588 patients (median age 61·0 years [IQR 55-66]; 124 [21%] female and 464 [79%] male) at 57 hospitals in China were randomly assigned to the serplulimab group (n=292) or placebo group (n=296). With a median follow-up duration of 42·7 months (IQR 24·3-53·6), median event-free survival was significantly longer with serplulimab than with placebo in the PD-L1 CPS ≥10 population (not reached [NR] vs 42·0 months; hazard ratio 0·65 [95% CI 0·47-0·90]; p=0·0082). With a median follow-up of 35·9 months (IQR 23·5-49·4), median event-free survival was also significantly longer with serplulimab than with placebo in the intention-to-treat population (NR vs 35·9 months; 0·73 [95% CI 0·56-0·94]; p=0·015). Grade 3 or worse treatment-related adverse events occurred in 136 (47%) patients in the serplulimab group and 172 (59%) patients in the placebo group; 19 (7%) and 31 (11%) patients in the respective groups discontinued treatment due to treatment-related adverse events. INTERPRETATION:Neoadjuvant serplulimab plus SOX followed by adjuvant serplulimab significantly improved event-free survival and demonstrated a better safety profile compared with neoadjuvant and adjuvant SOX in PD-L1-positive, resectable gastric or gastro-oesophageal junction adenocarcinoma. Extended follow-up for the overall survival data is warranted to confirm a survival advantage of this perioperative strategy with a chemotherapy-sparing adjuvant component for this indication. FUNDING:Shanghai Henlius Biotech.

INTRODUCTION:Assessing progress in cancer control at the national level is crucial, as it offers key insights into the overall effectiveness of the health system. Here, we aimed to describe 20-year trends in stage distribution, treatment patterns, and survival for gastric cancer in China, and to estimate stage- and treatment-specific 1-, 3-, 5-, and 10-year overall survival (OS) at the national level. METHODS:In this study, we included data from 40,158 patients diagnosed with gastric cancer between 1998 and 2022, derived from nine tertiary grade-A hospitals located across all four major regions of China. The characteristics, treatment modalities, and survival outcomes were analyzed across four predefined time periods: period 1 (1998-2005), period 2 (2006-2010), period 3 (2011-2015), and period 4 (2016-2022). Stage- and treatment-specific survival with 95 % confidence intervals (CIs) was estimated using the Kaplan-Meier method; differences were assessed by log-rank test. RESULTS:During the study period, there was an increasing proportion of patients with early stage gastric cancer from 9.8 % in 1998 to 19.5 % in 2022, while patients with metastatic disease remained stable. For early gastric cancer classified as cT1aN0, the proportion of patients undergoing endoscopic resection increased from 0.0 % in period 1-30.4 % in period 4. Among patients with locally advanced gastric cancer, the proportion receiving neoadjuvant therapy increased from 0.8 % to 17.3 %. Regarding survival outcomes, the 5-year OS rate of gastric cancer cohort increased from 38.2 % in period 1-46.1 % in period 4. Stage-specific 5-year OS rates were 90.0 % (stage I), 69.0 % (stage II), 37.0 % (stage III), and 13.7 % (stage IV). Compared with Japan, gastric cancer patients with stage IA in China exhibited higher 5-year OS rate (91.9 % vs. 89.6 %) but a much lower proportion (16.9 % vs. 43.6 %). Survival for stage IIIA-IIIC was poorer in China (48.5 % vs. 59.3 %, 34.7 % vs. 45.6 %, 21.8 % vs. 29.9 %; respectively). CONCLUSIONS:From 1998 to 2022, the proportion of patients diagnosed with early-stage gastric cancer in China gradually increased, and treatment strategies changed substantially over time, accompanied by a modest improvement in the 5-year OS rate. The disparities in stage distribution and survival outcomes between China and Japan highlight the need to further improve early detection of gastric cancer and to develop more effective treatment modalities for patients with stage III disease in China.

Gastric cancer is characterized by marked molecular heterogeneity, with TTN and TP53 mutations occurring in more than 50

BACKGROUND AND OBJECTIVE:Obesity and metabolic syndrome impose chronic lipotoxic stress on hepatocytes, contributing to macrophage reprogramming and fibrosis progression in steatotic liver disease. However, how soluble hepatocyte-derived factors regulate Kupffer cell-hepatic stellate cell (HSC) crosstalk remains unclear. This study investigated the role of hepatocyte NAT10-mediated N4-acetylcytidine (ac4C) RNA modification and explored potential ac4C-dependent targets involved in this process. METHODS:Hepa1-6 cells were exposed to palmitic acid with or without NAT10 knockdown, and nd key findings were validated in primary mouse hepatocytes. Hepatocyte-conditioned media were applied to primary Kupffer cells and a Kupffer-HSC coculture system. ac4C-RIP-qPCR and mRNA stability assays were performed. Recombinant TGFβ1 was used for rescue analysis. In vivo, hepatocyte-specific NAT10 knockdown was achieved using AAV8-TBG-shNat10 in a high-fat diet plus fructose model. RESULTS:NAT10 knockdown reduced TGFβ1 secretion from lipotoxic hepatocytes without affecting cell viability. Conditioned media induced Spp1 and Lgals3 expression in Kupffer cells, which was attenuated by NAT10 silencing. In the Kupffer-HSC coculture system, Kupffer cells primed by lipotoxic hepatocyte-conditioned media markedly increased HSC activation, as indicated by elevated α-SMA and Collagen I expression. This effect was diminished when Kupffer cells were exposed to media from NAT10-deficient hepatocytes, and recombinant TGFβ1 partially restored HSC activation. Tgfb1 showed reduced ac4C enrichment and mRNA stability following Nat10 knockdown, suggesting that it is a NAT10-responsive ac4C-associated transcript under lipotoxic conditions. In vivo, hepatocyte-specific NAT10 knockdown reduced hepatic TGFβ1 levels, decreased CLEC4F+LGALS3+ pro-fibrotic Kupffer cells, and alleviated liver fibrosis. CONCLUSIONS:Hepatocyte NAT10-mediated ac4C regulation contributes to pro-fibrotic Kupffer-HSC crosstalk in steatotic liver disease, at least partly through TGFβ1-associated soluble hepatocyte-derived signaling.

BackgroundThe tumor microenvironment (TME) of gastric adenocarcinoma is exceptionally heterogeneous, and epithelial-mesenchymal transition (EMT) is a central mechanism promoting local invasion and metastatic spread. Even so, how EMT-programmed cells are spatially arranged within tumor tissue, how they interact with neighboring immune populations, and which molecular nodes might be exploited therapeutically remain incompletely defined.MethodsWe built a large-scale, multi-layered analytical pipeline that combined single-cell transcriptomics (approximately 250,000 cells drawn from two independent patient cohorts), Visium-based spatial transcriptomics processed through Bayesian cell2location deconvolution, niche-level SpaTopic modeling, deep-learning histology analysis (ResNet50 feature extraction paired with CellProfiler-derived morphometrics), and ensemble survival modeling spanning over one hundred algorithmic combinations. Gaussian mixture modeling was used to define EMT-high cell states, the Scissor framework was applied to link fibroblast subsets to patient mortality, and a multi-tier filtering scheme was used to nominate druggable candidate genes. The top candidate, PVR/CD155, was interrogated experimentally by RT-qPCR, Western blotting, immunohistochemistry, and siRNA knockdown in AGS cells. We further evaluated PVR druggability by molecular docking, a 100-nanosecond molecular dynamics trajectory, and MM/GBSA binding-energy estimation using PP-121 as a candidate ligand.ResultsSub-clustering identified seven fibroblast subclusters: Fib_APOD, Fib_COL4A1, Fib_SLPI, Fib_COL1A1, Fib_CCL4, Fib_STMN1, and Fib_S100B. The SLPI-high subset preferentially localized to peritoneal metastases. Phenotype-guided Scissor mapping nominated a survival-associated fibroblast program enriched within COL4A1-expressing fibroblast states. PVR/CD155 was prioritized as an EMT-linked candidate gene; single-cell expression profiling showed that PVR was most frequently detected in endothelial, epithelial and fibroblast compartments, with low detection in lymphoid and plasma cells. PVR/CD155 was significantly upregulated in gastric cancer cell lines and tumor tissues. PVR/CD155 knockdown in AGS cells decreased proliferation, migration and invasion, supporting a tumor cell-intrinsic functional role rather than establishing PVR as a stromal immune biomarker. Molecular docking and dynamics simulations identified PP-121 as a candidate PVR-binding ligand for future biochemical validation.ConclusionThe proposed framework connects single-cell resolution with spatial and histological data, produces validated prognostic tools and nominates PVR/CD155 as a tumor cell-intrinsic EMT-associated target candidate for gastric cancer. Stromal or immune regulatory roles of PVR remain plausible but require direct compartment-specific and functional validation.

Background: Radiotherapy is crucial for managing esophageal squamous cell carcinoma (ESCC). This research explored the potential and mechanism of enhancing ESCC radiosensitivity through targeting phosphoglycerate kinase 1 (PGK1). Methods: After ESCC cells were exposed to X-rays and C-ions, hub genes were identified through proteomic analysis and bioinformatics. To elucidate PGK1’s function, small interfering RNAs and plasmids were used to silence and overexpress PGK1 in two human ESCC cell lines. Plate colony formation, cell counting kit 8, and 5-ethynyl-2′-deoxyuridine assays were conducted to detect cell proliferation after irradiation with different linear energy transfer rays (X-rays and carbon ions). Flow cytometry was used to assess radiation-induced perturbations in the cell cycle, apoptosis, reactive oxygen species (ROS), and mitochondrial membrane potential. Western blotting was performed to detect the protein expressions of protein kinase B (Akt), phosphorylated protein Kinase B (pAkt), mammalian target of rapamycin (mTOR), and phosphorylated mammalian target of rapamycin (pmTOR). Results: Proteomics and bioinformatics analyses revealed that PGK1 plays a key role in modulating ESCC radiosensitivity. Knockdown of PGK1 resulted in the suppression of cancer cell proliferation and viability, promoted apoptotic processes, and demonstrated a synergistic anti-tumor effect in conjunction with radiation. Conversely, overexpression of PGK1 promoted cancer cell growth and increased radiation resistance. This may be attributed to the accumulation of ROS and the inhibition of Akt/mTOR pathway following PGK1 inhibition. Conclusion: Targeting PGK1 may be an effective strategy to increase ESCC radiation sensitivity, offering a promising strategy for improving treatment outcomes.

BACKGROUND:This study aims to compare the efficacy between neoadjuvant immune checkpoint inhibitors (ICIs) plus chemotherapy vs . chemotherapy, and neoadjuvant triplet vs . doublet chemotherapeutic regimens in locally advanced gastric/esophagogastric junction cancer (LAGC). METHODS:We included LAGC patients from 47 hospitals in China's National Cancer Information Database (NCID) from January 2019 to December 2022. Using propensity score matching (PSM), we retrospectively analyzed the efficacy between neoadjuvant ICIs plus chemotherapy vs . chemotherapy alone, and neoadjuvant triplet vs . doublet chemotherapeutic regimens. The primary study result was the pathologic complete response (pCR) rate. The secondary study results were disease-free survival (DFS) and overall survival (OS). RESULTS:A total of 1205 LAGC patients were included. After PSM, the ICIs plus chemotherapy and the chemotherapy cohorts had 184 patients each, while the doublet and triplet chemotherapy cohorts had 246 patients each. The pCR rate (14.13% vs . 7.61%, χ2 = 4.039, P = 0.044), and the 2-year (77.60% vs . 61.02%, HR = 0.67, 95% con-fidence interval [CI] 0.43-0.98, P = 0.048) and 3-year (70.55% vs . 61.02%, HR = 0.58, 95% CI 0.32-0.93, P = 0.048) DFS rates in the ICIs plus chemotherapy cohort were improved compared to those in the chemotherapy cohort. No significant increase was observed in the OS rates at both 1 year and 2 years. The pCR rates, DFS rates at 1-3 years, and OS rates at 1-2 years did not differ significantly between the doublet and triplet cohorts, respectively. No differences were observed in postoperative complications between any of the group comparisons. CONCLUSIONS:Neoadjuvant ICIs plus chemotherapy improved the pCR rate and 2-3 years DFS rates of LAGC compared to chemotherapy alone, but whether short-term benefit could translate into long-term efficacy is unclear. The triplet regimen was not superior to the doublet regimen in terms of efficacy. The safety after surgery was similar between either ICIs plus chemotherapy and chemotherapy or the triplet and the doublet regimen.

BACKGROUND:NADPH oxidase 4 (NOX4) plays an important role in metabolic reprogramming, epithelial-mesenchymal transition (EMT), and other cellular processes by strictly regulating intracellular ROS generation. However, there is a lack of analysis on the role of NOX4 in the tumor immune microenvironment and predictive value for prognosis and immunotherapy response in various tumor types. METHODS:This study used data from the Cancer Genome Atlas (TCGA), Chinese Glioma Genome Atlas (CGGA), Tumor Immune Single-cell Hub (TISCH), Genotype-Tissue Expression (GTEx), cBioPortal, Tumor Immune Estimation Resource (TIMER 2.0), and ROC Plotter databases to analyze the expression, prognosis, biological function, immune cell infiltration, and genetic and epigenetic changes of NOX4 in various tumor types. Meanwhile, in vitro experiments were conducted to verify the role of NOX4 in the migration, invasion, proliferation, and apoptosis of glioblastoma (GBM). RESULTS:The findings indicated that NOX4 is upregulated in various types of cancer, accompanied by gene amplification, mutation, and deletion. The high expression of NOX4 is associated with poor prognosis in various cancers. Functional enrichment analysis showed that NOX4 is mainly enriched in immune and cancer progression pathways, such as angiogenesis, EMT, interferon response, inflammatory response. Immune cell infiltration analysis showed that high expression of NOX4 is associated with increased infiltration of cancer-associated fibroblasts (CAF) and macrophages. In vitro experiments showed that silencing NOX4 inhibits the proliferation, migration, and invasion of GBM and increases cell apoptosis. CONCLUSION:NOX4 can serve as a prognostic and immunotherapy response biomarker for various cancers and targeting NOX4 may be a feasible anti-tumor therapy and may have synergistic anti-tumor effects combined with immunotherapy.

ST6 β-galactoside α2,6-sialyltransferase 1 (ST6GAL1), a crucial enzyme for tumor-associated sialic acid modification, has been reported to positively correlate with colorectal cancer (CRC) tumorigenesis; however, the underlying mechanism remains unclear. To elucidate the protumor mechanisms of ST6GAL1, we performed transcriptomic and N-glycoproteomic analyses and in vitro assays. We found that ST6GAL1 was significantly upregulated in tumor samples than in matched normal samples by analyzing fresh clinical samples from public databases (mean mRNA expression level: tumor vs. normal samples = 0.002712:0.000966, P < 0.05, n = 22). The in vitro results revealed that ST6GAL1 overexpression promoted CRC cell proliferation, migration, and chemoresistance, which were significantly blocked by its knockdown. Transcriptomic data showed that many genes related to the four modules (proliferation/cell cycle, migration, motility, and epithelial-mesenchymal transition) were upregulated after ST6GAL1 overexpression but downregulated after ST6GAL1 knockdown. Furthermore, the N-glycoproteome data revealed that 25 substrates that were sialylated upon ST6GAL1 overexpression were related to protumor activity. Importantly, we found that knockdown of lectin galactoside-binding soluble 3-binding protein (LGALS3BP), a newly identified secreted substrate of ST6GAL1, significantly blocked the proliferation, invasion, and chemoresistance of CRC cells induced by ST6GAL1 overexpression. Treatment with sialidases (neuraminidases, NAs) also blocked the protumor activity of ST6GAL1. Thus, ST6GAL1-induced increased sialylation of substrates such as LGALS3BP and upregulation of protumor genes promote CRC tumorigenesis and chemoresistance, which provides important perspectives and new targets for the treatment of CRC.

Interferon gamma (IFNG) directly affects the antitumor immune response. ST6-β-galactoside α-2,6-sialyltransferase 1 (ST6GAL1) is also positively correlated with poor prognosis for colorectal cancer (CRC). we performed some works to exploreexplored the underlying mechanism to further understand immune escape in colorectal cancer (CRC). First, we used clinical samples to confirm the relationship between ST6GAL1 and CRC. Afterward, we constructed overexpression/knockdown cell lines and performed bulk RNA sequencing, kinase phosphorylation chip, mass spectrometry, and in vitro and in vivo assays to explore the mechanism regulated by ST6GAL1. Finally, we verified the mechanism by performing immunoprecipitation and immunofluorescence (IF) staining. ST6-β-galactoside α-2,6-sialyltransferase 1 (ST6GAL1) expression was negatively correlated with the response to neoadjuvant chemotherapy in patients with CRC and negatively correlated with the sensitivity to IFNG in CRC cell lines. We confirmed that ST6GAL1 overexpression inhibited the infiltration of effector T cells, the levels of IFNG produced by CD8+ T cells and CD4+ T cells decreased, and the levels of granzyme B produced by effector cells decreased in animal models. The results of in vitro assays and phosphorylation chip detection revealed that ST6GAL1 inhibited IFNG receptor 1 (IFNGR1) phosphorylation, thus decreasing the activation of the JAK1/STAT1 signaling pathway. Through immunoprecipitation assays and mass spectrometry, we found that ST6GAL1 can sialylate BICD cargo adaptor 2 (BICD2), thereby affecting the interaction between BICD2 and IFNGR1 to ultimately inhibit the phosphorylation of IFNGR1 and promote immune escape. Our results confirmed that ST6GAL1 decreases the sensitivity of tumor cells to IFNG. This study describes a novel mechanism by which ST6GAL1 promotes the immune escape and malignant progression of CRC.

Colorectal cancer (CRC), one of the most prevalent and lethal malignancies of digestive system, continues to impose a substantial burden on global health due to its high morbidity and mortality. Tumor microenvironment (TME) is a critical regulator for CRC progression and therapeutic response, but the in-depth understanding on the relationship of TME with CRC remains to be elucidated. In this study, we leveraged single-cell RNA sequencing (scRNA-seq) and bulk RNA-seq data to dissect the immune heterogeneity in CRC patients. The differential expression genes analysis, functional enrichment analysis, random forest analysis and the Least Absolute Shrinkage and Selection Operator method were used to construct a molecular immune prognostic model. The molecular model demonstrated robust performance in stratifying patients based on their immune microenvironment characteristics. The experimental results showed that TIMP1 was highly expressed in CRC. Knockdown of TIMP1 gene significantly inhibited RKO cell proliferation and invasion. By integrating scRNA-seq and bulk RNA-seq data, we developed a new prognostic model that effectively predicts clinical outcomes in patients with CRC and identifies TIMP1 as a promising prognostic biomarker for CRC.

BACKGROUND AND OBJECTIVE:Chromobox 2 (CBX2) plays a pivotal role in the malignant phenotypes of several cancers, which has been found to up-regulated in gastric cancer (GC). This study aimed to investigate the specific function and underlying mechanism of CBX2 in GC. METHODS:The potential role of CBX2 in GC was uncovered based on online bioinformatic analysis. The protein and mRNA expression levels were assessed by immunohistochemistry, qRT-PCR, and western blot. Using parental and the corresponding resistant GC cell lines, the effect and mechanism of CBX2 on 5-Fu resistance were explored by cell transfection technique, CCK-8 kit, colony formation assay, flow cytometry, the commercial kit, as well as co-IP. Xenograft tumor nude mice models were applied for in vivo assay. RESULTS:The expression of CBX2 was up-regulated in tumor tissues of GC patients and positively correlated with chemo-resistance, as well as that of EZH2. Knockdown of CBX2 resensitized 5-Fu resistant GC cells to 5-Fu while overexpressing CBX2 enhance GC cells against 5-Fu via the regulation of ferroptosis. In vivo experiments demonstrated that CBX2 overexpression enhanced 5-Fu sensitivity in tumors. Mechanically, CBX2 and EZH2 cooperatively suppressed ferroptosis in GC cells by inducing trimethylation of H3k27 (H3k27me3). Suppressing EZH2 blocked the inductive effect of CBX2 overexpression on 5-FU sensitivity of GC cells and reduced ferroptosis and H3k27me3 levels in CBX2 overexpressed GC cells. CONCLUSION:CBX2/EZH2 cooperatively confers 5-FU resistance in GC cells through suppressing ferroptosis via H3k27me3, which may lead to a promising therapeutic strategy for GC.

Phosphoglycerate kinase 1 (PGK1), a pivotal enzyme in the glycolysis pathway, contributes to tumor progression through diverse biological activities like cell metabolism, angiogenesis, proliferation, and epithelial-mesenchymal transformation (EMT). Although PGK1 has been intensively researched in specific cancer types, its overarching significance in pan-cancer contexts remains underexplored. This study leveraged various public database resources, including the Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx), Tumor Immune Estimation Resource (TIMER2.0), and cBioPortal, to analyze the gene expression, gene alteration characteristics, prognostic value, subcellular localization, biological function, immune characteristics, and drug sensitivity of PGK1 in 33 different cancer types. R software was used to visualize these data. Furthermore, the effects of PGK1 on the proliferation, apoptosis, migration, and invasion of esophageal squamous cell carcinoma (ESCC) cells were also examined in vitro using 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay, CCK-8 assay, Annexin V-FITC/PI assay, migration assay, and invasion assay. The findings suggested that PGK1 is upregulated in various cancer types and closely associated with poor prognosis. In terms of functional enrichment analysis, PGK1 primarily plays a role in glycolysis, hypoxia, EMT, and immune-related pathways. Furthermore, PGK1 is highly expressed in immune and malignant cells in the tumor microenvironment. Notably, PGK1 expression varied significantly among immune cells with distinct activation states. The results of experiments in vitro showed that PGK1 was significantly upregulated in ESCC cells, and its knockdown led to significant inhibition of proliferation, migration, and invasion while increasing cell apoptosis; conversely, overexpression promoted proliferation, migration, and invasion while reducing apoptosis. PGK1 can serve as a prognostic biomarker and therapy target for various cancers, and it may be a promising focal point of immunological studies.

Tumor-infiltrating B cells (TIBs) are the most important cell type involved in the immune response. TIBs display considerable intratumor heterogeneity due to genetic variation, epigenetic differences and transcriptional plasticity in the tumor microenvironment (TME). Owing to the unique anatomical location of CRC, the B cell subpopulation exhibits more extensive heterogeneity. Many studies have shown that TIBs have gradually become a key predictor of immunotherapy for malignant cancers, including CRC. TIBs have essential functions, including antigen presentation and antibody secretion, and they promote T-cell activation and myeloid chemotaxis. However, owing to the complex TME, TIBs not only promote the antitumor immune response but also inhibit the immune response. With the in-depth study of tumor-infiltrating T cells, tumor-associated myeloid cells and the interactions among these cells in the TME, the special role of immune cells in the TME has gradually become clear. However, the influence of TIBs in the TME and their interactions with nonimmune cells in the TME remain unclear. Here, we summarize the current progress in TIBs based on single-cell RNA sequencing in CRC in recent years, focusing on specific effector or regulatory characteristics of different B cell subclusters in the CRC TME.

Neutrophil elastase (NE) is a proteolytic enzyme released extracellular during the formation of neutrophil extracellular traps (NETs) through degranulation. In addition to participating in the body's inflammatory response, NE also plays an important role in cancer. It can promote tumor proliferation, migration, and invasion, induce epithelial-mesenchymal transition (EMT), and change the tumor microenvironment (TME) to promote tumor progression. Concurrently, NE promotes systemic treatment resistance by inducing EMT. However, it can also selectively kill cancer cells and attenuate tumor development. Sivelestat is a specific NE inhibitor that can be used in the perioperative period of esophageal cancer patients to reduce the incidence of postoperative complications after esophagectomy. In addition, the combination of sivelestat and trastuzumab can enhance the efficacy of human epidermal growth factor receptor 2(HER 2) positive breast cancer patients. Meanwhile, targeting the human antibody domains and fragments of NE is also a new way to treat cancer and inflammation-related diseases. This review provides valuable insights into the role of NE in cancer treatment. Additionally, we discuss the challenges associated with the clinical application of sivelestat. By shedding light on the promising potential of NE, this review contributes to the advancement of cancer treatment strategies.

Despite the exact biological role of HNF1 homolog A (HNF1A) in the regulatory mechanism of glioblastoma (GBM), the molecular mechanism, especially the downstream regulation as a transcription factor, remains to be further elucidated. Immunohistochemistry was used to detect the expression and clinical relevance of HNF1A in GBM patients. CCK8, TUNEL, and subcutaneous tumor formation in nude mice were used to evaluate the effect of HNF1A on GBM in vitro and in vivo. The correction between HNF1A and epidermal growth factor receptor pathway substrate 8 (EPS8) was illustrated by bioinformatics analysis and luciferase assay. Further mechanism was explored that the transcription factor HNF1A regulated the expression of EPS8 and downstream signaling pathways by directly binding to the promoter region of EPS8. Our comprehensive analysis of clinical samples in this study showed that upregulated expression of HNF1A was associated with poor survival in GBM patients. Further, we found that knockdown of HNF1A markedly suppressed the malignant phenotype of GBM cells in vivo and in vitro as well as promoted apoptosis of tumor cells, which was reversed by upregulation of HNF1A. Mechanistically, HNF1A could significantly activate PI3K/AKT signaling pathway by specifically binding to the promoter regions of EPS8. Moreover, overexpression of EPS8 was able to reverse the apoptosis of tumor cells caused by HNF1A knockdown, thereby exacerbating the GBM progression. Correctively, our study has clarified the explicit mechanism by which HNF1A promotes GBM malignancy and provides a new therapeutic target for further clinical application.

B7 -H3 is a common oncogene found in various cancer types. However, the molecular mechanisms underlying abnormal B7 -H3 expression and colorectal cancer (CRC) progression need to be extensively explored. B7 -H3 was upregulated in human CRC tissues and its abnormal expression was correlated with a poor prognosis in CRC patients. Notably, gain- and loss -of -function experiments revealed that B7 -H3 knockdown substantially inhibited cell proliferation, migration, and invasion in vitro, whereas exogenous B7 -H3 expression yielded contrasting results. In addition, silencing of B7 -H3 inhibited tumor growth in a xenograft mouse model. Mechanistically, our study demonstrated that the N6-methyladenosine (m6A) binding protein YTHDF1 augmented B7 -H3 expression in an m6A-dependent manner. Furthermore, rescue experiments demonstrated that reintroduction of B7 -H3 considerably abolished the inhibitory effects on cell proliferation and invasion induced by silencing YTHDF1. Our results suggest that the YTHDF1-m6A-B7-H3 axis is crucial for CRC development and progression and may represent a potential therapeutic target for CRC treatment.