Renal cell carcinoma (RCC) is a highly prevalent malignancy within the urinary system. While surgery combined with adjuvant therapy has significantly improved survival rates for early-stage patients, those with advanced disease or intolerance to surgery face significant therapeutic challenges. A core issue is the limited efficacy of radiotherapy, often constrained by the intrinsic radioresistance of tumors. As a crucial mechanism of epitranscriptomic regulation, m6A RNA methylation profoundly influences the malignant progression of RCC and the radioresistance of various cancers by dynamically regulating key gene networks. Targeting central components of the m6A regulatory pathway (e.g., METTL3, FTO, and YTHDF family) holds promise as a novel strategy to overcome therapeutic hurdles in RCC and enhance tumor radiosensitivity.

Over the past decades, advances in technique have turned radiotherapy into a precise and effective modality for cancer treatment. Nevertheless, the biological effect on organs surrounding the tumor target area is a main dose-limiting element in radiotherapy, e.g., doses are restricted by lethal intestinal tissue toxicity for many pelvic or abdominal cancer radiotherapy. Consequently, minimizing the damage to normal tissues has become a focal point in radiotherapy research. Recently, ionizing radiation delivered at ultra-high dose rates (also termed FLASH-RT) has emerged in the field. Preclinical studies in a variety of animal models have shown that FLASH-RT can control tumors while minimizing normal tissue toxicity compared to conventional radiotherapy (CONV-RT), which broadens the therapeutic window of radiotherapy. In this review, we summarize the present speculations on the biological mechanisms underlying the FLASH effects, including oxygen depletion hypothesis, mitochondria hypothesis, genome stability hypothesis, immunomodulation hypothesis, and so on. Furthermore, we summarize the main data supporting the clinical translation of FLASH-RT, and discuss its relevance and applicability in the treatment of digestive system tumors.

To develop and evaluate a intralesional and perilesional radiomics strategy based on different machine learning model to differentiate International Society of Urological Pathology (ISUP) grade > 2 group and ISUP ≤ 2 prostate cancers (PCa). 340 case of PCa patients confirmed by radical prostatectomy pathology were obtained from two hospitals. The patients were divided into training, internal validation, and external validation groups. Radiomic features were extracted from T2-weighted imaging, and four distinct radiomic feature models were constructed: intralesional, perilesional, combined tumoral and perilesional, and intralesional and perilesional image fusion. Four machine learning classifiers logistic regression (LR), random forest (RF), extra trees (ET), and multilayer perceptron (MLP) were employed for model training and evaluation to select the optimal model. The performance of each model was assessed by calculating the area under the ROC curve (AUC), accuracy, sensitivity, specificity, positive predictive value, negative predictive value, and F1 score. The AUCs for the RF classifier were higher than that of LR, ET, and MLP, and was selected as the final radiomic model. The nomogram model integrating perilesional, combined intralesional and perilesional, and intralesional and perilesional image fusion had an AUC of 0.929, 0.734, 0.743 for the training, internal, and external validation cohorts, respectively, which was higher than that of the individual intralesional, perilesional, combined intralesional and perilesional, and intralesional and perilesional image fusion models. The proposed nomogram established from perilesional, combined intralesional and perilesional, and intralesional and perilesional image fusion radiomic has the potential to predict the differentiation degree of ISUP PCa patients. Not applicable.

Objectives FLASH radiotherapy is garnering attention for its capacity to diminish skin toxicity without compromising tumoricidal efficacy, presenting a stark contrast to conventional (CONV) radiotherapy. Despite its promise, the underlying molecular mechanisms of FLASH irradiation (FLASH-IR) on skin are not yet fully elucidated. Methods This study investigated the transcriptomic responses of human foreskin fibroblast cells (HFF-1) via the FLASH-IR or CONV irradiation (CONV-IR), employing the next-generation RNA sequencing (RNA-seq) to capture the gene expression profiles. Our comparative analysis aimed to dissect the cellular and molecular pathways influenced by these two irradiation methods. Results We identified a spectrum of differentially expressed genes (DEGs), signaling pathways, and transcriptional networks that were either shared or divergent between FLASH-IR and CONV-IR. Particularly, transcription factor NR4A1 showed significant upregulation in response to FLASH-IR, while chromatin stability factor ELF3 was markedly downregulated following CONV-IR. The top 10 up-regulated DEGs were subjected to qPCR validation, confirming their differential expression in response to FLASH-IR and CONV-IR. Conclusion Collectively, our findings delineate unique regulatory landscapes of FLASH-IR and CONV-IR on skin cells, corroborating established effects and shedding new light on the molecular interplay within the context of ultra-high dose radiation.

FLASH radiotherapy (FLASH-RT) is a radiotherapy technique that achieves ultra-high dose rates in a fraction of a second. Based on data from experimental animal models, FLASH-RT appears to protect a number of normal tissues from radiation-induced damage, including the brain, gastrointestinal tract, and lung, while conventional radiotherapy (CONV-RT) causes radiation-induced toxicity in these tissues. In this review, we provide a brief summary of the history of radiation therapy and focus on some of the most recent FLASH-RT papers and findings. It is particularly noteworthy that pulmonary fibrosis represents a common complication of radiotherapy. New evidence indicates that FLASH-RT, unlike traditional radiotherapy methods, might help protect lung cancer patients from developing pulmonary fibrosis caused by radiation. FLASH-RT will advance more quickly than anticipated, although there are still a number of unresolved concerns. FLASH-RT will be a safer and more effective option for lung cancer treatment.

In recent years, there has been a steady increase in professionals engaged in radioactive work. The biological impacts of long-term exposure to low dose-rate radiation remain elusive, as there is a dearth of systematic research in this field. BEAS-2B cells were used to establish a cell model with continuous passaging after radiation exposure, which was subsequently subjected to in vivo tumorigenesis assays and in vitro malignant phenotype experiments. By scRNA-seq, we conducted copy number variation analysis, cell trajectory analysis, and cell communication analysis. Furthermore, we used FACS, molecular docking, multiplex immunohistochemistry, qRT-PCR, and co-immunoprecipitation to validate and further explore the molecular mechanisms driving tumor evolution. Long-term low dose-rate exposure is associated with a higher degree of malignancy, as evidenced by the induction of more CNV and EMT events, as well as the delayed activation of DNA repair pathways, which trigger increased genomic instability. The long-term low dose-rate specific ligand-receptor pair, ANGPTL4-SDC4, enhances cell malignancy by promoting angiogenesis in newly formed lung tumor cells. This study not only provides the first evidence and mechanistic explanation that long-term low dose-rate radiation leads to increased cellular malignancy but also offers valuable theoretical insights into the dynamic processes of early tumor evolution in lung cancer within the realm of tumor biology.

Aneuploidy is a hallmark of genomic instability that leads to tumor initiation, progression, and metastasis. CDC20, Bub1, and Bub3 form the mitosis checkpoint complex (MCC) that binds the anaphase-promoting complex or cyclosome (APC/C), a crucial factor of the spindle assembly checkpoint (SAC), to ensure the bi-directional attachment and proper segregation of all sister chromosomes. However, just how MCC is regulated to ensure normal mitosis during cellular division remains unclear. In the present study, we demonstrated that LNC CRYBG3, an ionizing radiation-inducible long noncoding RNA, directly binds with Bub3 and interrupts its interaction with CDC20 to result in aneuploidy. The 261–317 (S3) residual of the LNC CRYBG3 sequence is critical for its interaction with Bub3 protein. Overexpression of LNC CRYBG3 leads to aneuploidy and promotes tumorigenesis and metastasis of lung cancer cells, implying that LNC CRYBG3 is a novel oncogene. These findings provide a novel mechanistic basis for the pathogenesis of NSCLC after exposure to ionizing radiation as well as a potential target for the diagnosis, treatment, and prognosis of an often fatal disease.

We investigated the clinical implications and molecular features of TLS in stage I lung adenocarcinoma (LUAD).

BackgroundTumor abnormal protein (TAP), the sugar chain protein released by tumor cells during metabolism, allows the development of a technique that exploits aggregated tumor-associated abnormal sugar chain signals in diagnosing malignancies. Clinically, we have found that TAP detection can well predict some malignancies, but several physicians have not paid attention, and related studies have been minimal.MethodsWe evaluated TAP’s ability to distinguish between malignancies and benign diseases by receiver operating characteristic (ROC) curve analysis and studied the possibility of monitoring malignancy progression by evaluating TAP levels in follow-up. We used Kaplan-Meier survival curves and Cox proportional hazard regression models to investigate the relationship between TAP and prognosis.ResultsTAP levels were higher in whole solid malignancies and every type of solid malignancy than in benign patients. ROC curve analysis showed that TAP levels aid in distinguishing between malignancies and benign diseases. TAP levels decreased in patients with complete remission (CR) after treatment and increased in patients with relapse from CR. Patients with metastases had higher TAP levels than non-CR patients without metastases. There was no difference in overall survival among patients with different TAP levels, and multivariate analysis suggested that TAP was not an independent risk factor for solid malignancies.ConclusionTAP is an effective screening biomarker for many solid malignancies that can be used to monitor the progression of malignancies but not to prognosticate.

ObjectiveThis study aimed to evaluate the effectiveness of multi-phase-combined contrast-enhanced CT (CECT) radiomics methods for noninvasive Fuhrman grade prediction of clear cell renal cell carcinoma (ccRCC).MethodsA total of 187 patients with four-phase CECT images were retrospectively enrolled and then were categorized into training cohort (n=126) and testing cohort (n=61). All patients were confirmed as ccRCC by histopathological reports. A total of 110 3D classical radiomics features were extracted from each phase of CECT for individual ccRCC lesion, and contrast-enhanced variation features were also calculated as derived radiomics features. These features were concatenated together, and redundant features were removed by Pearson correlation analysis. The discriminative features were selected by minimum redundancy maximum relevance method (mRMR) and then input into a C-support vector classifier to build multi-phase-combined CECT radiomics models. The prediction performance was evaluated by the area under the curve (AUC) of receiver operating characteristic (ROC).ResultsThe multi-phase-combined CECT radiomics model showed the best prediction performance (AUC=0.777) than the single-phase CECT radiomics model (AUC=0.711) in the testing cohort (p value=0.039).ConclusionThe multi-phase-combined CECT radiomics model is a potential effective way to noninvasively predict Fuhrman grade of ccRCC. The concatenation of first-order features and texture features extracted from corticomedullary phase and nephrographic phase are discriminative feature representations.

Long noncoding RNAs (lncRNAs) in eukaryotic transcripts have long been believed to regulate various aspects of cellular processes, including carcinogenesis. Herein, it is found that lncRNA AFAP1-AS1 encodes a conserved 90-amino acid peptide located on mitochondria, named lncRNA AFAP1-AS1 translated mitochondrial-localized peptide (ATMLP), and it is not the lncRNA but the peptide that promotes the malignancy of nonsmall cell lung cancer (NSCLC). As the tumor progresses, the serum level of ATMLP increases. NSCLC patients with high levels of ATMLP display poorer prognosis. Translation of ATMLP is controlled by m6 A methylation at the 1313 adenine locus of AFAP1-AS1. Mechanistically, ATMLP binds to the 4-nitrophenylphosphatase domain and non-neuronal SNAP25-like protein homolog 1 (NIPSNAP1) and inhibits its transport from the inner to the outer mitochondrial membrane, which antagonizes the NIPSNAP1-mediated regulation of cell autolysosome formation. The findings uncover a complex regulatory mechanism of NSCLC malignancy orchestrated by a peptide encoded by a lncRNA. A comprehensive judgment of the application prospects of ATMLP as an early diagnostic biomarker for NSCLC is also made.

Objective:To investigate the differences in small intestinal toxicity and taxonomic composition,diversity,and functional pathways of gut microbiome and metabolome after different radiotherapies in mouse colorectal cancer(CRC)model.Methods:Azoxymethane/dextran sodium sulfate(AOM/DSS)-induced mouse CRC model was treated with single pulse FLASH-RT(dose rate 100 Gy/s)or CONV-RT(dose rate 2 Gy/min)at whole abdomen.At 12 d after radiotherapy,sections of small intestinal tract tissue were dissected for hematoxylin and eosin(HE)staining and the fresh feces were collected for 16S ribosomal RNA(rRNA)microbiome sequencing and liquid chromatography and mass spectrometry(LC-MS)metabolomics sequencing to assess changes in the gut microbiota and metabo-lites.Microbial high-throughput 16S rRNA data was analyzed with QIIME2 and LEfSe softwares.ProteoWizard,XCMS and Ropls softwares were used for LC-MS analysis.Results:HE staining showed that FLASH-RT maintained small intestinal integrity and reduced the radiotherapy-induced injury.Sequencing analysis of gut fecal microbiome showed that phylum Bacteroidetes and genera Prevotella and Lactobacillus of microbial community were increased after FLASH-RT.Metabolomics sequencing analysis revealed that the metabolites after FLASH-RT were enriched in amino acid metabolism,while cholesterol metabolism was top enriched after CONV-RT.Conclusions:FLASH-RT significantly mitigates the small intestine tissue damage compared with CONV-RT.FLASH-RT and CONV-RT have different impact on gut microbiota and its metabolites.Our results provide a theoretical basis for the early evaluation,prediction and individualized treatment of the irradiation effect after novel FLASH-RT on tumors through the evaluation of intestinal microbiota and metabolites.

Background: It remains undetermined whether preoperative computed tomography (CT)-guided hookwire localization would result in elevated risk of tumor spread through air spaces (STAS) in stage IA lung adenocarcinoma. Methods: A total of 1836 patients who underwent lobectomy were included. To eliminate the potential impact of confounding factors on producing STAS, propensity score–matching (PSM) was used to create two balanced subgroups stratified by implementation of hookwire localization. We also introduced an external cohort including 1486 patients to explore the effect of hookwire localization on the incidence of STAS and patient survival after sublobar resection (SR). For proactive simulation of hookwire localization, 20 consecutive lobectomy specimens of p-stage IA lung adenocarcinoma were selected. Results: Ex vivo tests revealed that mechanical artifacts presenting as spreading through a localizer surface (STALS) could be induced by hookwire localization but be distinguished by CD68 and AE1/3 antibody-based immunohistochemistry. The distance of STALS dissemination tended to be shorter compared with real STAS (P = 0.000). After PSM, implementation of hookwire localization was not associated with elevated STAS incidence, nor worse survival in p-stage IA patients undergoing lobectomy irrespective of STAS. Conclusions: CT-guided hookwire localization might induce mechanical artifacts presenting as STALS which could be distinguished by immunohistochemistry, but would not affect survival in p-stage IA disease. Surgeons can be less apprehensive about performing hookwire localization in relation to STAS on stage IA disease suitable for SR.

Backgrounds Prior investigations of the tumor microenvironment (TME) of diffuse large B-cell lymphoma (DLBCL) have shown that immune and stromal cells are key contributing factors to patients’ outcome. However, challenges remain in finding reliable prognostic biomarkers based on cell infiltration. In this study, we attempted to shed some light on chemokine C–C motif chemokine ligand 8 (CCL8) in DLBCL via interaction with M2 macrophages. Methods The Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data (ESTIMATE) algorithm was applied to evaluate immune and stromal scores from transcriptomic profiles of 443 DLBCL samples from The Cancer Genome Atlas (TCGA) and GSE10846 datasets. Immune cell infiltration (ICI) clusters were obtained based on different immune cell infiltrations of each sample, and gene clusters were derived through differentially expressed genes (DEGs) between the distinct ICI clusters. Five immune-related hub genes related to overall survival (OS) and clinical stages were obtained by COX regression analysis and protein–protein interaction (PPI) network construction then verified by quantitative real-time PCR (qPCR) and immunofluorescence staining in the FFPE tissues. The Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and TIMER websites were employed to explore the biological functions of CCL8-related DEGs. Uni- and multivariable Cox regression analyses were performed to analyze CCL8 as an independent prognostic risk factor in GSE10846 and were verified in other independent GEO cohorts. Results A higher stromal score was associated with favorable prognosis in DLBCL. Patients in the ICI B cluster and gene B clusters had a better follow-up status with a higher programmed death ligand 1 (PD-L1) and cytotoxic T-lymphocyte antigen 4 (CTLA4) expression. Most of ICI-related DEGs were enriched for immune-related signaling pathways. Five hub genes with a distinct prognosis association were identified, including CD163, which is a biomarker of M2 macrophages, and CCL8. Abundant M2 macrophages were discovered in the high-CCL8 expression group. The functional analysis indicated that CCL8 is a key component of immune-related processes and secretory granule groups. Cox regression analysis and data from other GSE datasets yielded additional evidence of the prognostic value of CCL8 in DLBCL. Conclusions CCL8 has been implicated in macrophage recruitment in several solid tumors, and only a few reports have been published on the role of CCL8 in the pathogenesis of hematological malignancies. This article attempted to find out TME-related genes that associated with the survival in DLBCL patients. CCL8 was identified to be involved in immune activities. Importantly, a series of bioinformatics analysis indicated that CCL8 might become an effective target for DLBCL, which interacts with M2 macrophage and immune checkpoint. The potential related mechanisms need to be further elucidated.

Traditional monolayer cell cultures often fail to accurately predict the anticancer activity of drug candidates, as they do not recapitulate the natural microenvironment. Recently, three-dimensional (3D) culture systems have been increasingly applied to cancer research and drug screening. Materials with good biocompatibility are crucial to create a 3D tumor microenvironment involved in such systems. In this study, natural silk fibroin (SF) and chitosan (CS) were selected as the raw materials to fabricate 3D microscaffolds; Besides, sodium tripolyphosphate (TPP), and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) were used as cross-linking agents. The physicochemical properties of obtained scaffolds were characterized with kinds of testing methods, including emission scanning electron microscopy, x-ray photoelectron spectroscopy, fourier transform infrared spectroscopy, water absorption, and swelling ratio analysis. Cancer cell lines (LoVo and MDA-MB-231) were then seeded on scaffolds for biocompatibility examination and drug sensitivity tests. SEM results showed that EDC cross-linked scaffolds had smaller and more uniform pores with great interconnection than the TPP cross-linked scaffolds, and the EDC cross-linked scaffold exhibited a water absorption ratio around 1000% and a swelling ratio of about 72%. These spatial structures and physical properties could provide more adhesion sites and sufficient nutrients for cell growth. Moreover, both LoVo and MDA-MB-231 cells cultured on the EDC cross-linked scaffold exhibited good adhesion and spreading. CCK8 results showed that increased chemotherapeutic drug sensitivity was observed in 3D culture compared with 2D culture, particularly in the condition of low drug dose (<1 μ M). The proposed SF/CS microscaffold can provide a promising in vitro platform for the efficacy prediction and sensitivity screening of anticancer drugs.

Transforming growth factor-β1 (TGF-β1) acts as a tumor promoter in advanced prostate cancer (PCa). We speculated that microRNAs (miRNAs) that are inhibited by TGF-β1 might exert anti-tumor effects. To assess this, we identified several miRNAs downregulated by TGF-β1 in PCa cell lines and selected miR-3691-3p for detailed analysis as a candidate anti-oncogene miRNA. miR-3691-3p was expressed at significantly lower levels in human PCa tissue compared with paired benign prostatic hyperplasia tissue, and its expression level correlated inversely with aggressive clinical pathological features. Overexpression of miR-3691-3p in PCa cell lines inhibited proliferation, migration, and invasion, and promoted apoptosis. The miR-3691-3p target genes E2F transcription factor 3 (E2F3) and PR domain containing 1, with ZNF domain (PRDM1) were upregulated in miR-3691-3p-overexpressing PCa cells, and silencing of E2F3 or PRDM1 suppressed PCa cell proliferation, migration, and invasion. Treatment of mice bearing PCa xenografts with a miR-3691-3p agomir inhibited tumor growth and promoted tumor cell apoptosis. Consistent with the negative regulation of E2F3 and PRDM1 by miR-3691-3p, both proteins were overexpressed in clinical PCa specimens compared with noncancerous prostate tissue. Our results indicate that TGF-β1-regulated miR-3691-3p acts as an anti-oncogene in PCa by downregulating E2F3 and PRDM1. These results provide novel insights into the mechanisms by which TGF-β1 contributes to the progression of PCa.