Background: Non-small cell lung cancer (NSCLC) remains a major cause of cancer mortality. Isoginkgetin, a natural biflavonoid, has shown pharmacological potential, yet its antitumor mechanisms in NSCLC are not well defined. Purpose: This study investigated the antitumor activity of isoginkgetin in NSCLC and explored its molecular targets and underlying mechanisms. Study design: Cell-based assays and in vivo xenograft models combined with transcriptomic profiling, redox an-alyses, and in silico target identification/validation. Methods: Cell-based assays were performed to evaluate the effects of isoginkgetin on NSCLC cell growth, migration, apoptosis, and epithelial-mesenchymal transition (EMT). Transcriptome profiling and redox analyses were conducted to assess mechanistic pathways. Target identification was conducted through in silico screening, followed by drug-target binding validation and functional characterization, including further validation through Methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) overexpression and silencing. The antitumor efficacy of isoginkgetin was further examined in NSCLC xenograft models, and the safety was assessed through acute toxicity experiments. Results: Isoginkgetin suppressed proliferation and migration, induced apoptosis, and blocked EMT in NSCLC cells. It disrupted redox homeostasis by enhancing oxidative stress, mediated by elevated reactive oxygen species (ROS) accumulation and activating endoplasmic reticulum (ER) stress and mitochondrial apoptosis. MTHFD2 was identified as a direct target of isoginkgetin, which binds to MTHFD2 and suppresses its functional role, ultimately leading to NADPH depletion and oxidative stress. Silencing MTHFD2 reproduced the effects of iso-ginkgetin, while overexpression of MTHFD2 partly attenuated these effects. In vivo, isoginkgetin markedly reduced tumor growth without systemic toxicity. Conclusion: Isoginkgetin exerts potent antitumor effects in NSCLC by targeting MTHFD2 and inducing redox imbalance, supporting its promise as a novel therapeutic candidate.

Long noncoding RNAs (lncRNAs) play crucial roles in regulating chromatin dynamics and gene expression, and their dysregulation is closely linked to tumorigenesis. However, their specific functions in gastric cancer (GC) remain poorly understood. Here, we found that lncRNA solute carrier family 16 member 1 antisense RNA 1 (SLC16A1-AS1) was markedly overexpressed in GC through integrated RNA sequencing (RNA-seq) analysis and validation in clinical tissues. High SLC16A1-AS1 expression correlated with advanced cancer stage, greater invasion depth, and poorer patient prognosis. Functional assays showed that SLC16A1-AS1 overexpression promoted GC cell proliferation, whereas knockdown inhibited proliferation in vitro and in vivo. Mechanistically, SLC16A1-AS1 interacted with the 5-methylcytosine (m5C) methyltransferase NOL1/NOP2/ SUN (NSUN2), enhancing m5C modification of GRP78 mRNA, which stabilized the transcript and increased GRP78 protein levels. Rescue experiments demonstrated that GRP78 overexpression reversed the proliferation-inhibitory effect of SLC16A1-AS1 depletion. These findings reveal that SLC16A1-AS1 drives GC cell proliferation via NSUN2-mediated m5C modification of GRP78 mRNA, suggesting a potential target for GC diagnosis and therapy.

Acute lung injury (ALI) is characterized by dysregulated pulmonary inflammation, in which alveolar macrophages (AMs) play a crucial role. The m⁶A reader protein YTHDF2, known to facilitate mRNA degradation, is implicated in inflammation; however, its specific function in ALI pathogenesis remains unclear. Using myeloid-specific Ythdf2 knockout mice subjected to cecal ligation and puncture (CLP)-induced ALI, we demonstrate that Ythdf2 deficiency significantly attenuates lung injury, as evidenced by reduced histopathological damage, pulmonary edema, and inflammatory cell infiltration, along with lower levels of pro-inflammatory cytokines (IL-6, TNF-α) in both bronchoalveolar lavage fluid and serum. Moreover, Ythdf2 deletion was accompanied by a substantial upregulation of Hmox1 protein expression in both lung tissues and AMs, an observation consistent with the known role of Ythdf2 in pulmonary hypertension. Furthermore, global m⁶A methylation levels and the expression of key methyltransferases (Mettl3, Mettl14) were elevated during ALI, coinciding with increased Ythdf2 expression. This upregulation of m⁶A regulators (YTHDF2, METTL3, METTL14) was also confirmed in pulmonary macrophages from human septic lungs. In conclusion, our findings support the concept that the myeloid-specific deletion of Ythdf2 ameliorates lung injury, an effect closely associated with the upregulation of Hmox1, highlighting Ythdf2 as a potential therapeutic target for ALI.

Platinum-based drugs exhibit potent anti-tumor efficacy but are limited by low bioavailability, severe toxicity, and resistance. Current therapeutic strategies lack effective solutions due to the unclear molecular mechanisms. Autophagy, with its dual protective/destructive roles, offers potential to enhance platinum-based chemotherapy, yet its clinical translation for optimizing therapeutic outcomes remains underexplored. To address this challenge, we screened a Natural Compounds Library (NCL) to identify low-toxicity agents that synergize with Cisplatin (Cis). Among 285 autophagy-related candidates, Catharanthine (CA) emerged as a specific autophagy activator model molecule that reduced toxicity and synergistically suppressed gastric cancer (GC) when combined with Cis. Mechanistically, CA promoted organ protection via endoplasmic reticulum stress (ERS)/AMPKα-dependent autophagy activation. The CA-Cis combination induced tumor-suppressive effects, including ERS, autophagosome accumulation, and cytoskeletal impairment in cancer cells. Conversely, CA-mediated autophagy protected normal cells, as AMPKα knockdown abolished this protection, resulting in DNA damage and apoptosis. These results highlight the dual autophagic flux regulation: tumor cells undergo destructive autophagy, while normal cells experience protective autophagy, establishing a favorable therapeutic balance. We confirmed that the CA-Cis combination activates autophagy through the AMPKα-ULK1 pathway in both tumor and non-tumor tissues and differentially regulates phosphorylation at serine 757 of ULK1, this differentiation can dramatically modulate autophagy activity, thereby mediating context-dependent dual outcomes of autophagic protection and detrimental effects. These findings elucidate a mechanism whereby CA enhances platinum efficacy by remodeling of autophagic homeostasis in organisms, providing a theoretical basis for optimizing platinum-based regimens. Our study bridges autophagy's dual functionality with clinical strategy, proposing the combination of specific autophagy activators as a promising approach to overcome platinum resistance and toxicity in GC treatment.

Basal-type muscle-invasive bladder cancer (BMIBC) is characterized by aggressive metastasis and poor prognosis but lacks molecularly defined therapeutic targets. Through integrative analyses of clinical cohorts and BBN-induced mouse models, we identified KRT14 as a core oncogenic driver that orchestrates the KRT14-IGF2BP1 signaling axis to promote BMIBC progression and lung metastasis. Mechanistically, residues K294 and E295 within the KH2 domain of IGF2BP1 specifically recognize conserved residues D226 and E227 within the nuclear export signal of KRT14. This direct interaction facilitates IGF2BP1-mediated cytoplasmic trafficking and auto-stabilization of its own mRNA, establishing a positive feedback loop that amplifies IGF2BP1-targeted pro-invasive gene expression. Functional disruption of this axis suppressed primary tumor progression and lung metastasis in BMIBC models. Collectively, our findings define the KRT14-IGF2BP1 axis as a previously unrecognized, potentially targetable vulnerability in BMIBC, whose inhibition may limit aggressive disease progression and inform future therapeutic strategies.

Abstract Malignant tumors are major diseases that seriously threaten human health, and the unmet clinical demand for treatments in the field of oncology has been persistently large. The research and development of new antineoplastic drugs has become a powerful means to address this demand. The purpose of this guideline is to provide a systematic overview and summary of clinical research on antineoplastic drugs in terms of study format, trial staging, mechanism of action, ethical review, trial process, patient needs, and the evaluation of efficacy and adverse events. It provides practical suggestions and references to aid the fundamental role of clinical research on antineoplastic drugs, i.e., to address clinical needs and maximize patient benefits.

OBJECTIVE:Colorectal cancer (CRC) remains one of the leading causes of cancer-related death worldwide due to the lack of effective therapies. Here we explored the clinical basis and therapeutic promise of the integrin-focal adhesion kinase (FAK)-dependent pathway for CRC. METHODS AND RESULTS:Our bioinformatic and histological analyses showed that FAK was markedly upregulated at both mRNA and protein and signaling levels in the two CRC patient cohorts. Particularly, the portion of carcinomas carrying active FAK (Y397phosphorylation) increased by threefold from stage I to III/IV tumors or metastatic lesions. Consistent with this clinic landscape, FAK inhibition via knockdown or chemical inhibitors suppressed tumor cell growth largely in the subset of CRC cell lines with low MYC expression. In contrast, the FAK inhibition was less effective in the cell line pool with high MYC expression. The resistance to FAK targeting diminished upon a co-inhibition of BRD4 via BET inhibitors. It coincided with an induction of cell cycle arrest at G1-S and G2-M phases, elevated apoptosis and chemosensitivity (paclitaxel and oxaliplatin), and impaired stemness. Mechanistically, the BET inhibitor induced an EMT-like phenotype, tilting tumor cell dependence toward the integrin-FAK axis. Moreover, inhibiting FAK alone or in combination with SRC or BRD4 markedly suppressed cell motility and the YAP or MYC activation, and restored the expression of the long isoform BRD4. Also, co-genomic/genetic dysregulations of FAK and YAP1 or SRC strongly correlated with poor disease-free patient survival. CONCLUSION:Overall, our study highlights the potent pro-malignant role of the integrin-FAK axis in CRC, fueling its targeting as a single agent or synthetic lethal-based therapy.

This study was conducted to investigate the relationship between STK11 genetic alterations and the outcomes of patients with metastatic EGFR mutant lung cancer. Clinical characteristics and genomic data were downloaded from the cBioPortal database. The information of the case with STK11 mutation was collected from Jiangyin People’s Hospital. Univariate and multivariate analyses were performed to distinguish the prognostic differences. Outcomes were analyzed before and after propensity score matching (PSM). A patient with STK11 mutation was insensitive to osimertinib and had an extremely poor prognosis. Further analysis showed that STK11 mutations had a strong mutual exclusion with EGFR mutations. A total of 960 patients with metastatic EGFR mutant lung adenocarcinoma were enrolled in the prognostic analysis. STK11 alternation was a significant predictor of worse outcomes in univariate or multivariate analyses. After PSM, patients with STK11 alternations still exhibited poor prognoses. Cell culture experiments also showed that the loss of STK11 could contribute to the resistance of osimertinib. Functionally, STK11 mutation was positively associated with metabolic signaling pathways and immune infiltrates negatively. Through drug screening, trametinib was identified to sensitize osimertinib in the STK11-deficient cell. This study found that STK11 genetic alterations portend a worse prognosis for patients with metastatic EGFR mutant lung cancer and led to osimertinib resistance potentially. MEK inhibitors could sensitize osimertinib in the STK11-deficient cell.

The impact of Helicobacter pylori (H. pylori) status on gastric cancer survival remains unclear. In this study, we conducted a prognostic analysis of 488 gastric cancer patients and performed single-cell RNA sequencing (scRNA-seq) on 18,717 T cells from six tumor samples with varying H. pylori statuses. Our findings revealed that gastric cancer patients with H. pylori infection had significantly longer survival times compared to those with negative H. pylori status. After unsupervised re-clustering of T cells based on scRNA-seq data, we identified ten CD4+ and twelve CD8+ clusters. Among them, four CD8+ T cell clusters exhibited distinct distributions based on H. pylori infection status. One cluster, marked by CXCL13, showed high levels of IFNG and GZMB in H. pylori-infected patients, while another cluster, which expressed immune suppression related genes like AREG and PTGER2, was predominantly comprised of cells from non-infected patients. High PTGER2 expression was significantly associated with worse prognosis in patients with high CD8 expression. These insights advance our understanding of H. pylori’s influence on T cell responses in gastric cancer, aiding in treatment and prognostic strategies.

Survivors of early-onset colorectal cancer (EOCRC, i.e., diagnosed before age 50) are likely to experience recurrence after completing treatment. In this international, multi-centric, phase I-II-III EDRN biomarker study, we identified a panel of tumor-derived biomarkers of EOCRC recurrence. We then trained and independently validated a machine learning model (XGBoost) to predict 5-year recurrence-free and overall survival (RFS and OS) of patients with stage I-III EOCRC. Patients with “low-risk” EOCRC demonstrated statistically higher rates of 2-, 5-, and 10 year RFS in both the training cohort (51.0 vs. 92.4%; 34.4% vs. 92.4%; 25.8% vs. 92.4%, respectively; p < 0.0001) and the validation cohort (78.9% vs. 100.0%; 75.0% vs. 100.0%; 75.0% vs. 100.0%, respectively; p = 0.0019). We also report a significant reduction in both over-treatment and missed recurrences compared to current clinically available options. This tissue-based, machine learning-powered assay was prognostic of long-term RFS and OS outcomes after curative-intent treatment of EOCRC (ENCORE was first registered on ClinicalTrial.gov [ID: NCT06271980] on February 15th, 2024).

AbstractChemoresistance poses a significant clinical challenge in the treatment of gastric cancer (GC), while its underlying molecular mechanisms are still not fully understood. Post-translational protein modification and abnormal activation of nuclear factor-kappa B (NF-κB) are critical regulators of tumor chemoresistance. This study investigates the role of TNF receptors-associated factors 6 (TRAF6) in 5-Fluorouracil (5-FU) resistant GC. Utilizing short hairpin RNA (shRNA) to suppress TRAF6 expression in 5-FU resistant GC cells across both in vivo and in vitro models, we observed a marked reduction in cell proliferation and tumor growth. Low expression of TRAF6 inhibited nuclear translocation of NF-κB-p65, which was achieved by promoting the expression of Interferon regulatory factor 3 (IRF3). Importantly, TRAF6, an E3 ubiquitin ligase, bound to the IRF3-Δ (SR + IAD) (1-190aa) domain, inducing Lys70 ubiquitination of IRF3 to regulate its protein stability, with ubiquitin K48 residue playing a crucial role in this process. In conclusion, our study reveals the mechanism by which the TRAF6/IRF3 axis decreases GC’s cells sensitivity to 5-FU by promoting nuclear translocation of NF-κB-p65, offering valuable insights into overcoming chemoresistance in GC.

Objective:To provide the most up-to-date data on the burden of malignant mesothelioma (MM) and the projections through 2029 in China. Methods:Data on patients diagnosed with MM from China during 1990-2019 were obtained from the Global Burden of Disease (GBD) 2019 database, including annual cases and deaths data and age-standardized rates of incidence, mortality, and disability-adjusted life-years (DALYs) associated with MM among different age groups. Temporal trends during 1990-2019 were analyzed by the Joinpoint regression models using 95% confidence interval (CI), while the projections through 2029 were calculated by the Bayesian age-period-cohort model. Data on the production and consumption of asbestos in China were obtained from the United States Geological Survey on Mineral Commodity Summaries during 1996-2023. Results:We observed a significant elevation in incident new cases and deaths over the last 3 decades, increasing from 1193 in 1990 to 2815 in 2019 for incident cases and from 1134 in 1990 to 2773 in 2019 for death cases. We found a roughly 6% increase in the proportion of incident cases for those aged >70 years (30% in 2019 versus 24% in 1990), while for the proportion of deaths similar elevation for those aged >70 years was found. Additionally, men had significantly higher DALYs due to MM across age groups compared with women. Asbestos consumption in China dramatically dropped since 2012 and reached the bottom in 2017 with 230 kilotons. By 2029, the projected age-standardized rate for incidence and mortality is expected to reach 1.2 per million for both. Conclusion:We found, for the first time using GBD data on the Chinese population, that the burden of MM has been significantly increasing in China over the last three decades and will continue to increase in the upcoming decade, suggesting an urgent need for a complete ban on chrysotile asbestos in China.

Background:While timely assessment of long-term survival for patients with bone cancer is essential for evaluation on early detection and prognosis level of treatment of bone cancer, those data are extremely scarce in China. We aimed to timely and accurately assess long-term survival for patients with bone cancer in Eastern China. Methods:Patients diagnosed with bone cancer during 2004 - 2018 from four cancer registries with high-quality data from Taizhou, Eastern China were included. Five-year relative survival (RS) of bone cancer patients was calculated by period analysis for overall and the stratification. We further predicted 5-year RS during upcoming 2019 - 2023 using a model-based period analysis and survival data during 2004 - 2018. Results:Overall, 5-year RS for patients with bone cancer during 2014 - 2018 reached 46.6%, being 40.8% for male and 51.0% for female. Five-year RS declined along with aging, decreasing from 58.9% for age < 45 years to 41.5% for age > 60 years, while 5-year RS for urban area was higher compared to rural area (59.1% vs. 44.3%). The 5-year RS during upcoming 2019 - 2023 reached 48.3%. We found a clear upward trend in 5-year RS during 2004 - 2023 for overall and the stratification by sex, age at diagnosis, and region. Conclusions:We found that, for first time in China using period analysis, most up-to-date 5-year RS for patients with bone cancer reached 46.6% during 2014 - 2018, and is projected to reach 48.3% for the period 2019 - 2023, which has important implications for timely evaluation on early detection and prognosis level of treatment for patients with bone cancer in Eastern China.

The KCNQ family of genes ( KCNQ1- KCNQ5), encoding voltage-gated K + (Kv) channels, have been demonstrated to play potential pathophysiological roles in cancers. However, the associations between genetic variants located in KCNQ family genes and gastric cancer survival remain unclear. In this study, a large-scale cohort comprising 1135 Chinese gastric cancer patients was enrolled to identify genetic variants in KCNQ family genes associated with overall survival (OS). Based on the survival evaluation of all five KCNQ family genes, KCNQ1 was selected for subsequent genetic analysis. In both Cox regression model and stepwise Cox regression model used to evaluate survival-related genetic variants, we found that KCNQ1 rs10832417G>T was associated with an increased OS in gastric cancer patients (adjusted hazards ratio [HR] = 0.84, 95% confidence interval [CI]: 0.72-0.98, P = 0.023). Subsequently, a nomogram was constructed to enhance the prognostic capacity and clinical translation of rs10832417 variants. The rs10832417 T allele was predicted to increase the minimum free energy of the secondary structure. Furthermore, we observed that gastric cancer patients with downregulated KCNQ1 expression had a poorer survival across multiple public datasets. The findings of the present study indicate that KCNQ1 rs10832417 may serve as an independent prognostic predictor of gastric cancer, providing novel insights into the progression and survival of the disease.

Background The evolutionarily conserved protein FBXO9 acts as a substrate receptor for the SKP1-cullin-1-RBX1 ubiquitin ligase and is implicated in cancer, exhibiting either tumor-suppressive or oncogenic effects depending on the specific tumor type. However, their role in lung cancer metastasis remains unclear. Methods Lentiviral vectors carrying miRNA-based shRNA sequences for gene-specific knockdown were generated, and Lenti-CRISPR-Cas9 vectors containing gene-specific sgRNA sequences were designed. Gene overexpression was achieved using doxycycline-inducible lentiviral constructs, while gene knockdown or knockout cells were generated using shRNA and CRISPR-Cas9, respectively. Functional assays included migration, clonogenic survival assays, tumor sphere assays, and protein interaction studies using mass spectrometry, immunoprecipitation, and immunoblot analysis. Results This study identified FBXO9 as a crucial regulator that suppresses lung cancer cell migration, tumor sphere growth and restricts metastasis. We showed that FBXO9 facilitates the ubiquitination of the catalytic subunit A (ATP6V1A) of the Vacuolar-type H + -ATPase (V-ATPase), resulting in its interaction with the cytoplasmic chaperone HSPA8 and subsequent sequestration within the cytoplasm. This process hinders the assembly of functional V-ATPase, resulting in reduced vesicular acidification. In contrast, depletion of FBXO9 reduced ATP6V1A ubiquitination, resulting in increased V-ATPase assembly and vesicular acidification, thus promoting pro-metastatic Wnt signaling and metastasis of lung cancer cells. Furthermore, we demonstrated the effectiveness of inhibitors targeting V-ATPase in inhibiting lung cancer metastasis in a mouse model. Finally, we established a correlation between lower FBXO9 levels and poorer survival outcomes in patients with lung cancer. Conclusion These findings collectively elucidate the critical role of FBXO9 in regulating V-ATPase assembly and provide a molecular basis for FBXO9’s function in inhibiting lung cancer metastasis. This highlights the potential therapeutic opportunities of FBXO9 supplementation.

<p>Mutually exclusive CIs for the AA-IR combined treatment for PC3 cells and PZ cells at indicated doses.</p>

Background: Recently, the molecular classification of gastric cancer (GC) promotes the advances of GC patients' precision therapy and prognosis prediction. According to the Asian Cancer Research Group (ACRG), GC is classified as microsatellite instable (MSI) subtype GC, microsatellite stable/epithelial-to-mesenchymal transition (MSS/EMT) subtype GC, MSS/TP53- subtype GC, and MSS/TP53+ subtype GC. Due to the easy metastasis of EMT-subtype GC, it has the worst prognosis, the highest recurrence rate, and the tendency to occur at a younger age. Therefore, it is curious and crucial for us to understand the molecular basis of EMT-subtype GC. Methods: The expression of RHOJ was detected by quantitative real-time PCR (qPCR) and immunohistochemistry (IHC) in GC cells and tissues. Western blotting and immunofluorescence (IF) were conducted to examine the effects of RHOJ on the EMT markers' expression of GC cells. The GC cells' migration and invasion were investigated by transwell assay. The tumor growth and metastasis were demonstrated correspondingly in different xenograft models. Results: Firstly, it was noticed that RHOJ was significantly upregulated in EMT-subtype GC and RHOJ has close relationships with the EMT process of GC, based on the Gene Expression Omnibus (GEO) and the Cancer Genome Atlas (TCGA) databases. Next, transwell assay and tail vein metastasis models were conducted to verify that RHOJ mediates the EMT to regulate the invasion and metastasis of GC in vitro and in vivo. In addition, weakened tumor angiogenesis was observed after RHOJ knockdown by the angiogenesis assay of HUVEC. RNA-seq and further study unveiled that RHOJ aggravates the malignant progression of GC by inducing EMT through IL-6/STAT3 to promote invasion and metastasis. Finally, blocking the IL-6/STAT3 signaling overcame RHOJ-mediated GC cells' growth and migration. Conclusions: These results indicate that the upregulation of RHOJ contributes to EMT-subtype GC invasion and metastasis via IL-6/STAT3 signaling, and RHOJ is expected to become a promising biomarker and therapeutic target for EMT-subtype GC patients.