PURPOSE:Based on Monte Carlo simulations and biological experiments, this study aimed to quantitatively analyze the spatial distribution and complexity of DNA double-strand breaks (DSBs) induced by carbon-ion radiation. METHODS AND MATERIALS:Using X-rays as the reference radiation, γ-H2AX immunofluorescence assays combined with Monte Carlo simulations were performed to evaluate the spatial distribution and complexity of DNA damage in HeLa cells exposed to radiation with different linear energy transfer (LET) values. To analyze the number and size distribution of fluorescent foci, the biological experiments used radiation doses of 0.5 Gy, 1 Gy, and 2 Gy, respectively. Monte Carlo simulations were conducted using Geant4-DNA and TOPAS. DNA damage clusters were quantitatively analyzed using an improved dose-weighted DBSCAN algorithm. RESULTS:Experimental results showed that X-rays primarily induced small, spatially dispersed DNA lesions, whereas high-LET carbon ions caused dense, overlapping complex damage. The simulated and experimental results matched well for X-rays, but carbon ion irradiation showed a nonlinear correlation between damage clusters and foci size. Further analysis indicated that carbon ion-induced damage involving both "intra-track" and "inter-track" mechanisms. At equal doses, higher particle fluence in shallow regions led to more inter-track damage, compensating for lower single-particle radiation quality. A dose-dependent Ratio Factor was proposed and validated to correct the simulated and experimental data, as described by the regression equation: Ratio Factor = (0.74 ± 0.03) × Dose + (0.67 ± 0.01). CONCLUSIONS:This study clarified the spatial distribution characteristics of carbon-ion-induced DNA damage under various radiation conditions, and proposed a novel correction model for accurate prediction of DNA damage clusters, potentially applicable to carbon-ion radiotherapy.

Objective.Carbon ion lattice radiation therapy (CLRT) lacks systematic parameter selection strategies. This study aimed to develop an automated lattice placement system for CLRT and elucidate the impact of irradiation strategies (IS), lattice structures (Ls), vertex diameters, and gaps on relative biological effectiveness-weighted dose (RWD) and dose-averaged linear energy transfer (LETd) distributions.Approach.A proprietary MATLAB-based automated lattice placement system was applied to 15 bulky tumors using single-field optimization to mitigate range uncertainties. Two IS, lattice-only and lattice combined with a uniform background dose to the clinical treatment volume (lattice + CTV), were investigated. Lattice configurations included body-centered cubic and hexagonal close-packed structures, diameters of 10, 15, and 20 mm, and vertex gaps of 15, 20, 25, and 30 mm. Plan quality was assessed using the evaluation metrics of spatially fractionated radiation therapy, gross tumor volume, and organs at risk (OAR). Generalized estimating equations (GEEs) were employed to quantify the relative contribution of each parameter. Moreover, a multi-criteria decision framework integrating the technique for order of preference by similarity and a significance-based winning vote was constructed to identify recommended parameter configurations across varying clinical priorities.Main results.GEE regression indicated that the interaction between the IS and gap (26.86%), the main effect of the IS (17.55%), and gap (16.88%) were the primary contributors to variations in RWD and LETddistributions. Conversely, Ls had minimal effect (<2.5%). In both strategies, a 10 mm vertex diameter demonstrated the most favorable performance. However, the recommended gap diverged significantly. For lattice-only strategy, a 20 mm gap emerged as the recommended configuration, achieving target coverage while minimizing OAR doses. In contrast, for lattice + CTV strategy, the addition of a background RWD rendered a 30 mm gap the recommended choice.Significance.The proposed automated lattice placement system and irradiation-strategy-specific parameter selection strategy provide a robust quantitative foundation for enhancing the clinical adaptability and efficacy of CLRT.

BACKGROUND:Image-guided particle therapy (IGPT) has significantly advanced in recent years, particularly in the context of proton therapy. However, imaging solutions for carbon-ion radiotherapy (C-ion RT) remain limited. PURPOSE:This study introduces sliding-gantry cone-beam computed tomography (CBCT) and dual-panel digital radiography (DR) systems, both mechanically independent of carbon-ion delivery nozzles. We aim to evaluate the image quality metrics and verify the positioning accuracy of the imaging systems. METHODS:Image quality was evaluated in terms of spatial resolution, low contrast resolution, image uniformity, and effective imaging area using a multi-purpose imaging phantom, Catphan 700 phantom, and ImageJ software. The influences of planning computed tomography (CT) slice thicknesses (1-5 mm), radiation quality settings (90-130 kV), and registration algorithms (bony, grayscale, and fiducial marker registrations) on positioning accuracy were assessed using anthropomorphic head-neck and thoracoabdominal phantom images. The clinical feasibility of both systems was validated in 22 enrolled patients. RESULTS:The CBCT exhibited a lower in-plane spatial resolution (2.50 line pairs per millimeter (lp/mm)) than DR (2.80 lp/mm). Spatial resolution of the CBCT system was measured at 0.90 lp/mm using the CTP 714 module of the Catphan 700 phantom. Both systems achieved a low contrast resolution of 2.30%. DR provided superior image uniformity (1.12%-1.40%) compared with CBCT (2.20%). The effective imaging areas were comparable between the CBCT and DR systems (99.30%-99.50%). Positioning accuracy varied with planning CT slice thicknesses, radiation quality settings, and registration algorithms, showing mean translation displacements ranging from 0.01 to 0.48 mm. CBCT achieved inter-fraction translational positioning errors within 2 mm in 42.3% (22/52) of fractions and rotational positioning errors within 2° in 80.8% (42/52) of fractions, and DR achieved 33.8% (24/71) and 73.2% (52/71), respectively. CONCLUSION:The developed CBCT and DR systems achieved superior image quality and sub-0.5 mm positioning accuracy. These findings support the clinical feasibility of integrating CBCT and DR imaging systems into the C-ion RT workflow.

ObjectiveTo analyze the inhibitory effect of carbon ion (C-ion) radiation-induced biological effects on the proliferation of lung adenocarcinoma (LUAD) by multi-omics integration.MethodsWe investigated C-ion radiation effects on LUAD cellular responses using A549 and LLC cell lines. Clonogenic survival assays, DNA damage, and metastasis quantified radiation sensitivity, while a multi-cell co-culture system (A549/Beas-2B/LLC/MLE-12) differentiated direct vs. bystander effects. Integrated transcriptomics and targeted metabolomics identified radiation-responsive genes/metabolites, with pathway analysis conducted through MetaboAnalyst 6.0. Clinical relevance of CPT1, GCH1, and EPAS1 was assessed using UCSC Xena and Kaplan-Meier Plotter survival data. Radiation-induced molecular changes were validated by RT-qPCR and immunoblotting across cell types. An A549 tumor-bearing mouse model was established, and the growth and tumor size of the tumor-bearing mice were observed after irradiation with C-ions. Blood was taken from mice after anesthesia and necropsy to detect changes in the major differential metabolism factor arachidonic acid (AA), and tumor tissues were examined to detect changes in the expression of CPT1, GCH1, and EPAS1 in the tissues.ResultsC-ion irradiation exerts a dual anti-proliferative effect on lung adenocarcinoma cells: it directly induces DNA damage (increased γ-H2AX/53BP1 foci), suppresses clonogenic survival, and inhibits tumor cell migration and invasion (p < 0.05); meanwhile, it amplifies the bystander effect from co-cultured normal cells through metabolic reprogramming, thereby enhancing overall cytotoxicity. Transcriptomic-metabolomic integration identified CPT1, GCH1, and EPAS1 as central regulators of radiation-induced metabolic suppression, with AA, 3-hydroxytetradecanoic acid, and 2-methylglutaric acid constituting critical downstream mediators. Database analysis revealed that the differential expression of CPT1, GCH1, and EPAS1 was correlated with the prognostic survival status of patients with lung adenocarcinoma. Carbon ion irradiation downregulated the expression of GCH1 and upregulated CPT1 and EPAS1. Animal experiments demonstrated that carbon ions markedly inhibited tumor proliferation in tumor-bearing mice. The level of arachidonic acid in mouse blood was significantly increased (p < 0.05). Carbon ion radiation suppressed GCH1 expression and promoted the expression of CPT1 and EPAS1 in tumor tissues (p < 0.05).ConclusionC-ion irradiation suppresses lung adenocarcinoma through a dual mechanism involving direct induction of cellular DNA damage and a metabolically enhanced bystander effect, driven by the CPT1/GCH1/EPAS1 regulatory axis, with arachidonic acid serving as a key downstream mediator.

ObjectiveTo investigate the role of PCBP1 in the inhibition of lung adenocarcinoma proliferation by carbon irradiation.MethodsA549 cells were irradiated with different doses of carbon ions to observe clonal survival and detect changes in cell proliferation. Whole transcriptome sequencing and the Illumina platform were used to analyze the differentially expressed genes in A549 cells after carbon ion irradiation. The relationship between the expression levels of PCBP1, ACSL4, and ALOX15 and survival was analyzed by combining data from the UCSC database and the Kaplan–Meier Plotter public platform. Additionally, the knockdown of the poly (rC)-binding protein 1 (PCBP1) gene using siRNA techniques was employed to further investigate the relationship between the expression levels of PCBP1 and ALOX15. To investigate the relationship between ALOX15 expression and survival, we assessed changes in key indicators of ferroptosis (mitochondrial morphology, ROS, MDA, and divalent iron) in A549 cells after knocking down the PCBP1 gene using siRNA technology. Additionally, the expressions of PCBP1, ACSL4, and ALOX15 in different groups were further analyzed through RT-PCR and Western blot techniques. The differential expression of PCBP1, ACSL4, and ALOX15 in NSCLC tissues was found to correlate with clinical prognosis for survival.ResultsCarbon ions significantly inhibited the proliferation of A549 cells, and 5.16 Gy carbon ions significantly induced the expression of differentially expressed genes in these cells. Additionally, carbon ions inhibited the expression of PCBP1, which led to alterations in mitochondrial morphology in lung adenocarcinoma cells. This was associated with a significant increase in the levels of ROS, MDA, and Fe2+. Furthermore, low expression of PCBP1 promoted ferroptosis by increasing the expression of ACSL4 and ALOX15.ConclusionCarbon ions decreased the expression of PCBP1 in A549 cells, and low expression of PCBP1 inhibited tumor proliferation by promoting ferroptosis.

UBE2C (Ubiquitin-conjugating enzyme E2C) has been confirmed to be closely associated with the progression of various cancers, but its specific role and clinical diagnostic and prognostic value in prostate cancer (PRAD) remain unclear. This study systematically assessed the expression characteristics, prognostic significance, and genetic mutations of UBE2C in cancer patients by integrating data from databases such as TCGA, GEO, cBioPortal, and COSMIC. Experimentally, we explored the biological functions of UBE2C in the occurrence and development of PRAD using various methods, including functional enrichment analysis, CCK8 cell proliferation assay, colony formation assay, Transwell migration and invasion assay, Edu staining, 3D tumor spheroid culture, cell cycle analysis, apoptosis detection, and xenograft tumor models. After knocking down UBE2C expression, the proliferation ability, migration and invasion ability of PRAD cells, as well as the growth of xenograft tumors, were all inhibited, and the cell cycle process and apoptosis were changed accordingly. These findings provide favorable experimental evidence and theoretical support for UBE2C as a novel molecular marker for prognosis assessment in solid tumors.

Objective.In-beam positron emission tomography (PET) is a promising approach for dose monitoring in carbon-ion radiotherapy (CIRT). This study evaluated the feasibility of a dual-head PET system using lutetium-yttrium oxyorthosilicate (LYSO) scintillators and silicon photomultiplier (SiPM) detectors incorporating time-of-flight (TOF) technology to monitor carbon-ion beam dose distribution.Approach.The dual-head PET system includes four modules per head, each comprising a 12 × 12 array of LYSO crystals (4.14 mm × 4.14 mm × 20 mm) coupled with SiPM detectors. TOF information was integrated into image reconstruction using a TOF-maximum likelihood expectation-maximization algorithm to enhance spatial resolution and reduce noise, especially along the panel-to-panel (X-axis) direction. Two configurations were used: (1) a 4 × 1 module for single-energy beams (120.12-280.56 MeV u-1) and multi-energy beams (178.87, 190.19, 201.03 MeV u-1), and (2) a 2 × 2 module for 20 mm × 20 mm field measurements using a 151.32 MeV u-1beam. During the 5 s spill-off in-beam phase, the system detected 511 keVγ-rays from positron annihilation while suppressing promptγ-ray interference. Analysis included 48 in-beam periods of 261.03 MeV u-1carbon ions, divided into eight groups, and monitor units (MUs) estimates were based on the PET image peak intensity.Main results.In-beam PET imaging showed peak depth increased with beam energy, from 29.9 mm (120.12 MeV u-1) to 132.0 mm (280.56 MeV u-1). For multi-energy beams, three distinct peaks corresponded to different energies (178.87, 190.19, and 201.03 MeV u-1). The 20 mm × 20 mm field measurements showed full width at half maximum values of 20.42 mm (X-axis) and 19.76 mm (Y-axis), with a 45.6 mm peak depth (Z-axis) consistent with the single-energy results. PET-based MU estimation with a 261.03 MeV u-1beam showed decreasing relative error with more beam periods, reaching a stable value around 1% after 12 periods.Significance.In-beam PET offers a robust solution for dose monitoring and verification in CIRT, supporting precise dose delivery and better patient outcomes.

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.

Proton beam therapy (PBT) has been gradually introduced for treating choroidal melanoma. This study systematically reviewed clinical reports to evaluate the efficacy and safety of PBT in choroidal melanoma patients. This systematic review included all the primary studies involving PBT for choroidal melanoma patients through April 2024. Four publicly accessible databases were searched, and the statistical data were analyzed using STATA 15.0. The outcomes of interest included overall survival (OS), metastasis-free survival, local control rate, and adverse reactions. A total of six case series involving 1059 patients with choroidal melanoma were included. The random effect model meta-analysis showed that the 2-, 3-, 5-, and 10-year OS rates of patients with choroidal melanoma treated with PBT were 97

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.

ObjectiveThe use of proton beam therapy (PBT) for treating ocular conjunctival malignancies is on the rise across numerous medical centers. This study conducts a systematic review and meta-analysis to assess the effectiveness and safety of PBT in treating malignant conjunctival tumors.MethodsWe searched for studies on PBT for ocular conjunctival malignancies in PubMed, Embase, Cochrane Library, and Web of Science (WoS) databases up to November 25, 2023. Studies were selected and data were extracted by two independent reviewers based on pre-established inclusion and exclusion criteria. The quality of evidence was assessed using the GRADE method. Meta-analysis was performed using STATA version 16.0.ResultsAn initial search yielded 586 articles, from which six retrospective case series studies were selected involving 291 patients with ocular conjunctival malignancies, including 240 cases of conjunctival melanoma and 51 cases of conjunctival squamous cell carcinoma (SCC). Meta-analysis with a random-effects model showed that PBT is effective and relatively safe, with 2-, 4-, and 5-year overall survival (OS) rates of 98% (95% CI 95–102%), 87% (95% CI 69–104%), and 78% (95% CI 70–87%) respectively. Reported toxicity rates included 19% for cataracts, 10% for glaucoma, 5% for lacrimal stenosis, 52% for sicca symptoms, and 11% for limbal stem cell deficiency. The GRADE assessment yielded a low certainty of evidence.ConclusionsProton therapy offers a viable alternative treatment for patients with conjunctival malignancies, with acceptable treatment-related toxicity rates.

Radiotherapy is a vital treatment agent for lung adenocarcinoma (LUAD) patients, while radioresistance remains a major factor in treatment failure. Here, we aimed to elucidate how signal transducer and activator of transcription 1 (STAT1) affected sensitivity to carbon ion irradiation for LUAD cells in vivo and in vitro. The results of colony formation, CCK-8, EdU, and calcein-AM/PI double-staining assays demonstrated that the overexpression of STAT1 markedly enhanced the inhibitory effect of carbon ion irradiation on the viability of LUAD cells (A549 and PC9 cells). Lactate dehydrogenase (LDH) leakage assays identified ferroptosis as the predominant form of cell death induced by STAT1 overexpression in LUAD cells. Meanwhile, the ferroptosis-related PCR array confirmed heme oxygenase 1 (HO-1) as a potential effector molecule of STAT1-induced ferroptosis. Mechanistically, STAT1 overexpression resulted in phosphorylation at the serine 727 residue, triggering the upregulation of HO-1 expression and subsequent labile iron pool (LIP) accumulation. This process amplified the Fenton reaction, leading to increased reactive oxygen species (ROS), lipid peroxides (LPO), and glutathione (GSH) depletion. HO-1 knockdown eliminated the ferroptosis induced by the overexpression of STAT1. Furthermore, in vivo experiments showed that STAT1 overexpression enhanced the effect of carbon ion irradiation in inhibiting the growth of subcutaneous tumors in nude mice. These findings provide the foundation for the development of the STAT1-HO-1 axis as a radiosensitization target for LUAD patients.

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.

Purpose: Head and neck adenoid cystic carcinoma (HNACC) is a radioresistant tumor. Particle therapy, primarily proton beam therapy and carbon-ion radiation, is a potential radiotherapy treatment for radioresistant malignancies. This study aims to conduct a meta-analysis to evaluate the impact of charged particle radiation therapy on HNACC. Methods: A comprehensive search was conducted in Pubmed, Cochrane Library, Web of Science, Embase, and Medline until December 31, 2022. The primary endpoints were overall survival (OS), local control (LC), and progression-free survival (PFS), while secondary outcomes included treatment-related toxicity. Version 17.0 of STATA was used for all analyses. Results: A total of 14 studies, involving 1297 patients, were included in the analysis. The pooled 5-year OS and PFS rates for primary HNACC were 78% (95% confidence interval [CI] = 66-91%) and 62% (95% CI = 47-77%), respectively. For all patients included, the pooled 2-year and 5-year OS, LC, and PFS rates were as follows: 86.1% (95% CI = 95-100%) and 77% (95% CI = 73-82%), 92% (95% CI = 84-100%) and 73% (95% CI = 61-85%), and 76% (95% CI = 68-84%) and 55% (95% CI = 48-62%), respectively. The rates of grade 3 and above acute toxicity were 22% (95% CI = 13-32%), while late toxicity rates were 8% (95% CI = 3-13%). Conclusions: Particle therapy has the potential to improve treatment outcomes and raise the quality of life for HNACC patients. However, further research and optimization are needed due to the limited availability and cost considerations associated with this treatment modality.

Objectives: Cancer cells with ‘stemness’ are generally resistant to chemoradiotherapy. This study aims to compare the differences in radiation sensitivity of A549 and CD44+A549 stem-like cells to X-rays and carbon ion radiation (C-ions), and to find a target that can kill cancer stem-like cells (CSCs) of non-small cell lung cancer (NSCLC). Methods: The study used two cell lines (A549 and CD44+A549). The tumorigenicity of cells was tested with animal experiments. The cells were irradiated with X-rays and C-ions. Cell viability was detected using the CCK-8 and EdU assay. A liquid chromatograph-mass spectrometer (LC–MS) helped detect metabolic differences. Protein and mRNA expression were detected using a Western blot, reverse transcription-quantitative (RT-qPCR), and PCR array. The autophagic activity was monitored with a CYTO-ID® Autophagy Detection Kit 2.0. Immunofluorescence and co-immunoprecipitation helped to observe the localization and interaction relationships. Results: First, we verified the radio-resistance of CD44+A549 stem-like cells. LC-MS indicated the difference in autophagy between the two cells, followed by establishing a correlation between the radio-resistance and autophagy. Subsequently, the PCR array proved that TGM2 is significantly upregulated in CD44+A549 stem-like cells. Moreover, the TGM2 knockdown by small interfering RNA could decrease the radio-resistance of CD44+A549 cells. Bioinformatic analyses and experiments showed that TGM2 is correlated with the expression of CD44 and LC3B. Additionally, TGM2 could directly interact with LC3B. Conclusions: We established the CD44-TGM2-LC3 axis: CD44 mediates radio-resistance of CD44+A549 stem-like cells through TGM2 regulation of autophagy. Our study may provide new biomarkers and strategies to alleviate the radio-resistance of CSCs in NSCLC.

Background: Gliomas are characterized by aggressive behavior, leading to severe disability and high mortality. Ubiquitin-like modifier activating enzyme 2 (UBA2) is a subunit of the E1-activating enzyme involved in the SUMOylation (SUMO, small ubiquitin-related modifier) of numerous proteins. Although the abnormality of UBA2 is linked to the progression of various tumor types, the role of UBA2 in glioma is still unknown. Methods: A bioinformatic analysis using several public databases was conducted to examine the expression level, clinicopathological correlations, and prognostic significance of UBA2 in glioma. The correlation between UBA2 expression and drug sensitivity in cancers was also explored. Multiple cellular experiments were conducted to validate the role of UBA2 in glioma. Results: Analysis of multiple databases and cellular experiments revealed that UBA2 was overexpressed in glioma tissues and cell lines, respectively. UBA2 expression in gliomas correlated with World Health Organization (WHO) grade, IDH gene status, 1p19q deletion, histological type, and immune cell infiltration in glioma. UBA2 expression in carcinomas also correlated with drug sensitivity. Kaplan-Meier analysis revealed that high expression of UBA2 predicted poorer survival in glioma patients. A nomogram model containing UBA2 expression was constructed for clinical practice. Knockdown of UBA2 was observed to suppress glioma cell progression and sensitize glioma cells to irradiation in vitro. Conclusion: Overall, this research showed that UBA2 might be involved not only in the development of glioma but also in the regulation of immunity, drug sensitivity, and radiosensitivity. Therefore, UBA2 may be a potential target for therapy and a candidate biomarker for glioma diagnosis and prognosis.

Purpose: This systematic review and meta-analysis sought to assess whether ultra-high dose rate (UHDR) ion irradiations can induce the FLASH effect in animal models. Methods: A comprehensive search of the Web of Science, PubMed, and EMBASE databases was conducted from inception until March 20, 2023, to identify studies involving irradiated animals subjected to proton or carbon ion beams at varying dose rates. The research content should include various indicators that can reflect the effect and safety of radiation, such as survival, normal tissue toxicity, inflammatory response, tumor volume, etc Results: Compared to conventional dose rate (CONV) ion irradiations, UHDR ion irradiations can significantly improve mouse survival (HR 0.48, 95% CI 0.29 to 0.78, I2 = 0%) and maintain comparable tumor control. There was no significant impact of different dose rates on the survival of zebrafish embryos (SMD 0.11, 95% CI -0.31 to 0.53, I2 = 85%). Subgroup analysis showed that radiation dose was an important factor affecting the survival of zebrafish embryos. Achieving normal tissue sparing may require higher radiation dose under UHD.In mouse and zebrafish embryo models, normal tissue sparing did not always occur after UHDR ion irradiations. In addition, only a limited number of cytokines (CXCL1, IL-6, GM-CSF, G-CSF, HMGB1, and TGF-beta) and immune cells (microglia and myeloid cells) showed differences at different dose rates. Conclusions: UHDR ion irradiation can achieve FLASH effect, but the reproducibility of normal tissue sparing remains a challenge. Compared to CONV irradiation, UHDR ion irradiations demonstrated equivalent or even superior tumor control.

Background and goal: Carbon ion beam is radio -biologically more efficient than photons and is beneficial for treating radio -resistant tumors. Several animal experiments with tumor -bearing suggest that carbon ion beam irradiation in combination with immunotherapy yields better results, especially in controlling distant metastases. This implies that carbon ion induces a different anti -tumor immune response than photon beam. More complex molecular mechanisms need to be uncovered. This in vivo and in vitro experiment was carried out in order to examine the radio -immune effects and the mechanism of action of carbon ion beam versus X-ray in combination with PD -1 inhibitors. Methods and Materials: Lewis lung adenocarcinoma cells and C57BL/6 mice were used to create a tumor -bearing mouse model, with the non -irradiated tumor growing on the right hind leg and the irradiated tumor on the left rear. 10Gy carbon ion beam or X-ray radiation, either alone or in combination with PD -1 inhibitor, were used to treat the left back tumor. The expression of molecules linked to immunogenicity and the infiltration of CD8+ T lymphocytes into tumor tissues were both identified using immunohistochemistry. IFN- beta in mouse serum was measured using an ELISA, while CD8+ T cells in mouse peripheral blood were measured using flow cytometry. Lewis cells were exposed to different dose of X-ray and carbon ion. TREX1, PD -L1, and IFN- beta alterations in mRNA and protein levels were identified using Western blot or RT-PCR, respectively. TREX1 knockdown was created by siRNA transfection and exposed to various radiations. Using the CCK8 test, EdU assay, and flow cytometry, changes in cell viability, proliferation, and apoptosis rate were discovered. Results: Bilateral tumors were significantly inhibited by the use of carbon ion or X-ray in combination with PD -1, particularly to non -irradiated tumor(p<0.05). The percentage of infiltrating CD8+ T cells and the level of IFN- beta expression were both raised by 10Gy carbon ion irradiation in the irradiated side tumor, although PD -L1 and TREX1 expression levels were also elevated. Lewis cell in vitro experiment further demonstrated that both X-ray and carbon ion irradiation can up -regulate the expression levels of PD -L1 and TREX1 with dose -dependent in tumors, particularly the trend of up -regulation TREX1 is more apparent at a higher dose in carbon ion, i.e. 8 or 10Gy, while the level of IFN- beta is decreased. IFN- beta levels were considerably raised under hypofractionated doses of carbon ion radiation by gene silencing TREX1. Conclusions: By enhancing tumor immunogenicity and increasing CD8+T infiltration in TME through a threshold dosage, X-ray or carbon ion radiation and PD -1 inhibitors improve anti -tumor activity and cause abscopal effect in Lewis lung adenocarcinoma-bearing mice. TREX1 is a possible therapeutic target and prognostic marker.