Search Results (5,360)

Background/Objectives: Long non-coding RNAs (lncRNAs) are important regulators of tumor biology through their interactions with DNA, proteins, and non-coding RNAs. Although ENST00000615487.1 (also known as CTD-2396E7.11/AC010503.4) has been associated with multiple malignancies, its biological role in colorectal cancer (CRC) remains poorly characterized. This study aimed to investigate the expression pattern, cellular and subcellular localization, and potential functional role of ENST00000615487.1 in CRC using integrated in vitro and in silico approaches. Methods: Molecular characteristics of the transcript were obtained with the CPC2 and RNA Analyzer 3 tools. Differential expression of ENST00000615487.1 across 10 tumor types was analyzed using the UCSC Xena Browser. Transcript expression was experimentally evaluated in normal, tumor, and fibroblastic colon cell lines by PCR, while subcellular localization was assessed through the lncATLAS, lncLocator, and iLoc-LncRNA tools, and experimentally confirmed by qRT-PCR. Single-cell RNA sequencing data from the GSE161277 dataset were analyzed to determine cell type-specific expression patterns. Potential interactions with DNA, miRNAs, and proteins were investigated using Fasim-LongTarget, miRDB, and AnnoLnc2, followed by functional enrichment analyses using STRING and Enrichr. Results: ENST00000615487.1 was identified as a structurally stable non-coding transcript with a highly organized secondary structure. Differential expression analysis demonstrated significant downregulation in CRC compared with that in normal colon tissue. Single-cell transcriptomic analysis revealed predominantly epithelial-specific expression. In silico and experimental analyses demonstrated predominant nuclear localization in normal colon cells, whereas cytoplasmic enrichment was observed in CRC cells. Functional analyses identified potential interactions with HIP1R, RPH3AL, specific miRNAs, and proteins involved in transcriptional regulation and RNA processing pathways, as well as functional connections with proteins involved in vesicular transport. Conclusions: ENST00000615487.1 is a structurally stable lncRNA exhibiting context-dependent expression and localization patterns in CRC, suggesting a potential shift from nuclear transcriptional regulation toward cytoplasmic post-transcriptional functions during colorectal carcinogenesis. Full article

Anthraquinone-based intercalating compounds, such as doxorubicin and mitoxantrone, have long been used clinically due to their ability to induce DNA damage. More recently, heteroarene-fused anthraquinones have been developed to further enhance their anticancer activity. Among these compounds, 4,11-bis(2-(2-chloroacetamidine)ethylamino)anthra[2,3-b]thiophene-5,10-dione dihydrochloride (designated as derivative a) was identified as a potent apoptotic inducer. Based on this scaffold, two additional derivatives were synthesized by replacing the sulfur atom within the heterocyclic ring with nitrogen (derivative b) or oxygen (derivative c). Building upon our previous identification of ENOX2 as the primary target of this scaffold, the present study investigated the antiproliferative effects and underlying mechanisms of these derivatives in colon cancer cells with varying p53 statuses. Derivatives a and b effectively induced apoptosis and suppressed proliferation in p53 wild-type HCT116 cells, which was concomitantly accompanied by significant ENOX2 downregulation and the activation of intrinsic apoptotic signaling. In contrast, p53-null HCT116 cells exhibited reduced sensitivity, attenuated apoptotic responses, and minimal ENOX2 downregulation. Notably, derivative c primarily induced G2/M arrest rather than apoptosis regardless of p53 status, indicating a predominantly cytostatic mechanism. Collectively, these findings suggest that the degree of ENOX2 modulation is linked to the distinct anti-proliferative responses induced by heteroarene-fused anthraquinones, and that p53 status serves as a critical molecular switch influencing the transition between cytostatic growth arrest and apoptotic cell death. Full article

Introduction: For colon cancer (CC), KRAS mutations are associated with worse outcomes. Cytoreductive surgery (CRS) with hyperthermic intraperitoneal chemotherapy (HIPEC) for advanced-stage CC may proffer some survival benefits for select patients. We sought to evaluate postoperative morbidity and mortality among mutant and wildtype KRAS patients undergoing CRS/HIPEC for advanced CC. Methods: We identified 80 patients with CC from an institutional HIPEC database. Patients were categorized based on the presence (mutKRAS) or absence (wildtype) of KRAS mutations. Patient characteristics, tumor factors, treatment, and clinical outcomes were compared between groups. Postoperative morbidity was measured by Clavien–Dindo Classification (CDC). Kaplan–Meier survival analysis evaluated recurrence-free survival (RFS). Results: Overall, 52 (65%) mutKRAS were identified. Patient and tumor characteristics were similar among groups. While not significant, the wildtype group had a lower median (IQR) preoperative PCI score of 7 (3, 12) than the mutKRAS group, whose score was 11 (5, 21) (p = 0.057). The mutKRAS group had a higher rate of CDC grade 3 or more (23.1%) compared to the wildtype group (7.1%) (p = 0.015). Despite this, there were no significant differences in length of stay. Moreover, RFS between the mutKRAS and wildtype groups did not differ significantly (p = 0.47). Conclusions: Contrary to the typical association of KRAS mutations with poorer survival in CC, our findings suggest that patients with this mutation have an RFS comparable to those without it when treated with CRS/HIPEC. This may imply that CRS/HIPEC offers therapeutic benefits for KRAS-mutant CC patients, potentially mitigating the usually adverse prognosis associated with this mutation. Full article

Background: The highly heterogeneous and dynamically evolving tumor microenvironment leads to the development of drug resistance and recurrence in traditional therapies. Although immunotherapy demonstrates unique advantages, its clinical utility remains constrained by the suboptimal immunogenicity and the limited effect of monotherapy. Herein, a hybrid drug delivery system based on a spatiotemporal hierarchical controlled-release strategy was proposed to achieve dual immunotherapy with immune checkpoint blockade (ICB) and immunogenic cell death (ICD) to promote synergistic anti-tumor therapy. Methods: A liposome–micelle hybrid drug delivery system (RUT-RPP-LP) was constructed using a lipid bilayer composed of dioleoyl phosphatidylethanolamine/hemisuccinyl cholesterol to encapsulate rutin (RUT) and to form an inner cavity-encapsulated resveratrol micelle (RPP). RUT-RPP-LP was characterized, and its pH sensitivity and release behavior were investigated. Subsequently, a colon cancer tumor-bearing mouse model was constructed to evaluate the in vivo targeted anti-tumor effect and biological safety. On this basis, the combined mechanism of ICB and ICD was preliminarily explored. Results: RUT-RPP-LP possessed excellent formulation characteristics, stability, and biocompatibility, achieving graded controlled release of drugs via responding to the TME and lysosomal acidity, respectively. Obviously, RUT-RPP-LP could specifically target the tumor site, induce the occurrence of ICD, and simultaneously block the PD-1/PD-L1 immune checkpoint signaling pathway, thereby enhancing the function of T cells and inducing apoptosis of tumor cells. Conclusions: The RUT-RPP-LP based on the hierarchical controlled-release strategy exerted a spatiotemporally coordinated enhancement of anti-tumor immunity, and may provide a novel combinatorial approach to overcome the low response of immunotherapy in solid tumors. Full article

The vitamin D receptor (VDR) has been implicated in tumor progression, but its functional role in pancreatic ductal adenocarcinoma (PDAC) metastasis remains unclear. Here, using pharmacological modulation, gain- and loss-of-function approaches, transcriptomic profiling, and an experimental lung colonization model, we demonstrate that VDR acts as a suppressor of PDAC metastasis. Pharmacological activation of VDR by calcipotriol did not affect tumor cell proliferation but markedly inhibited the migratory and invasive capacities of multiple PDAC cell lines. In contrast, genetic deletion or pharmacological inhibition of VDR significantly enhanced metastatic phenotypes. To investigate the underlying mechanisms, we performed RNA sequencing on PDAC cells with differential VDR expression following calcipotriol treatment. Pathway enrichment analysis identified MAPK/ERK signaling as one of the most prominently altered pathways upon VDR activation. Functional studies further demonstrated that ERK inhibition abrogated the pro-metastatic effects induced by VDR loss or inhibition. In vivo lung colonization assays confirmed that VDR deficiency markedly promoted pulmonary metastatic colonization. Collectively, these findings identify VDR as a critical suppressor of PDAC cell metastatic colonization and reveal a previously unrecognized VDR–ERK regulatory axis that may represent a potential therapeutic target for limiting metastatic progression in pancreatic cancer. Full article

Background: Mucinous adenocarcinoma (MAC) of the colorectum is a biologically distinct histological subtype whose prognostic significance may vary substantially according to primary tumor location. The impact of anatomical site on clinicopathological characteristics and long-term survival in MAC remains incompletely characterized. This study aimed to evaluate the influence of tumor location on oncological outcomes in patients with mucinous colorectal cancer using a large population-based dataset. Methods: Patients diagnosed with mucinous colorectal adenocarcinoma between 2000 and 2023 were identified from the Surveillance, Epidemiology, and End Results (SEER) database. Operated patients were stratified by anatomical location into three groups: right colon (cecum, ascending colon, hepatic flexure, transverse colon), left colon (splenic flexure, descending colon, sigmoid colon, rectosigmoid junction), and rectum. The primary endpoints were overall survival (OS) and cancer-specific survival (CSS). Survival analyses were performed using the Kaplan–Meier method with log-rank testing. Multivariable Cox proportional hazards regression was used to assess the independent association of tumor location with survival outcomes, adjusting for age at diagnosis, year of diagnosis, sex, AJCC (American Joint Committee on Cancer) stage, tumor grade, receipt of radiotherapy and chemotherapy. Temporal trends in survival were evaluated across four consecutive diagnostic periods: 2000–2005, 2006–2011, 2012–2017, and 2018–2023. Results: A total of 40,698 patients with mucinous colorectal cancer were identified, of whom 40,174 underwent cancer-directed surgery (right colon n = 25,317; left colon n = 10,408; rectum n = 4449). The right colon was the predominant site of disease (62.9%). Median age was highest in the right colon group (74.0 years) and lowest in the rectum (65.0 years), while female sex predominated in right-sided tumors (55.7%) and male sex in rectal tumors (61.1%). Chemotherapy and radiotherapy utilization were markedly higher in the rectal group (68.6% and 64.5%, respectively) compared with the right colon (28.8% and 1.1%). Median OS was equivalent in the right and left colon groups (87.0 months each) but declined to 80.0 months in the rectal group. All pairwise CSS comparisons were statistically significant (all p < 0.001). On multivariable analysis, using the right colon as reference, the adjusted hazard ratios for CSS were 1.33 (95% CI: 1.28–1.39) for the left colon and 1.45 (95% CI: 1.34–1.57) for the rectum (both p < 0.001). Despite receiving the highest rates of multimodal therapy, rectal MAC demonstrated the worst long-term CSS across all anatomical groups. Temporal analyses revealed consistent CSS improvements in right-sided and left-sided MAC over the study period, whereas rectal MAC showed a non-linear trajectory with a plateau in the most recent diagnostic cohort (2018–2023). Conclusions: Mucinous colorectal adenocarcinoma demonstrates substantial biological and prognostic heterogeneity according to anatomical tumor location. Right-sided MAC was the most prevalent subtype and exhibited superior cancer-specific survival despite older patient age and lower treatment intensity. Rectal MAC demonstrated the worst long-term outcomes despite high utilization of multimodal neoadjuvant therapy, consistent with the established reduced responsiveness of mucinous tumors to conventional chemoradiotherapy. These findings underscore the necessity of incorporating anatomical location and tumor biology into individualized risk stratification and therapeutic planning for patients with mucinous colorectal cancer. Full article

Emerging evidence suggests that the microbiota may influence cancer metastasis and has been associated with multiple stages of the metastatic cascade. This review synthesizes recent advances using a unified framework centered on the three main stages of metastasis: invasion, circulation, and colonization. We highlight evidence linking several microbial taxa, including Fusobacterium nucleatum, enterotoxigenic Bacteroides fragilis, and Escherichia coli, to metastatic progression, and discuss how they may promote metastasis through distinct mechanisms in specific contexts. We further summarize stage-specific evidence indicating that the microbiota may contribute to local invasion, hematogenous dissemination, and distant colonization, offering a perspective on microbiota-associated metastatic progression. Finally, we discuss major limitations of the current literature, including the preponderance of preclinical and correlative evidence, constraints on causal inference, cancer-type and model dependence, low-biomass contamination, and open questions in interpreting tumor- and blood-derived microbial signals. Full article

Background/Objectives: Colorectal cancer commonly develops through an adenoma–carcinoma sequence, but the immune alterations accompanying this transition remain incompletely understood. Because we previously observed reduced γδ T cells, increased apoptosis, and frequent microsporidia infection in colorectal cancer, we investigated whether similar changes are already present in colonic adenomas. Methods: We studied 55 subjects, including 30 patients with colonic adenomas, 10 with colorectal cancer, and 15 healthy controls. Peripheral blood T-cell subsets and apoptosis were assessed by flow cytometry, tissue expression of IL-7, CD127, and CD132 was analyzed by RT-PCR, and microsporidia were detected in colonic tissues by immunofluorescence and real-time PCR. Results: γδ T cells were progressively reduced in colonic adenomas and further decreased in colorectal cancer, while apoptosis of both αβ and γδ T cells increased compared with healthy subjects. Tissue expression of CD132 was significantly downregulated in adenomas and colorectal cancer, whereas IL-7 expression was increased, particularly in adenomatous tissue, consistent with a compensatory mechanism attempting to preserve T-cell homeostasis despite reduced CD132 expression. Microsporidia prevalence rose from healthy controls to patients with colonic adenomas and colorectal cancer, and CD132 expression was more markedly reduced in microsporidia-positive colonic adenoma and cancer tissues. Conclusions: These findings indicate that altered IL-2 receptor-related signaling, γδ T-cell depletion, and increased apoptosis are already present at the adenoma stage and are associated with microsporidial infection. This pattern suggests a potential link between parasitic colonization and local immune dysfunction during colorectal tumorigenesis. Full article

Volatile organic compounds (VOCs) reflect cellular metabolic activities and may serve as non-invasive biomarkers in oncology. This study investigated whether acquired oxaliplatin resistance in colorectal cancer is associated with distinct volatilomic alterations. The human colorectal cancer cell line HCT116 and its oxaliplatin-resistant derivative (OXrHCT116) were analyzed under basal conditions and following oxaliplatin exposure. Chemoresistance was confirmed using dose–response, cell viability, and colony formation assays. VOCs were analyzed by headspace needle trap extraction coupled with gas chromatography–mass spectrometry (HS-NTE-GC-MS). OXrHCT116 cells exhibited markedly reduced oxaliplatin sensitivity, increased IC₅₀ values, and reduced proliferative and clonogenic capacity. Volatilomic profiling identified 55 significantly altered VOCs. Parental HCT116 cells displayed broader VOC diversity and higher turnover than OXrHCT116 cells. Hydrocarbons associated with lipid peroxidation and oxidative stress were more abundant in HCT116 cells, whereas resistant cells showed markedly reduced emission of these compounds. Additional alterations in aldehydes, alcohols, and aromatic compounds suggested reduced metabolic flux in resistant cells. Oxaliplatin exposure induced pronounced volatilomic changes in HCT116 cells but only minimal modulation in OXrHCT116 cells. These findings suggest that oxaliplatin resistance may be associated with distinct metabolic reprogramming and support VOC profiling as a promising approach for monitoring chemoresistance. Full article

Intestinal dysbiosis is a hallmark of both inflammatory bowel conditions and colorectal cancer, yet the mechanisms by which inflammatory mediators alter microbial communities and may contribute to tumor development remain poorly understood. Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine involved in innate immunity and the progression of inflammatory and neoplastic disorders. In this study, sequencing of the microbial 16S rRNA gene was performed to characterize the fecal microbiota profiles of wild-type (WT) and MIF-knockout (MIF-KO) BALB/c mice subjected to AOM/DSS-induced colitis-associated colorectal cancer (CAC). CAC induction resulted in marked microbial shifts, including increases in Muribaculaceae and Bacteroidota, in both WT and MIF-KO mice. Notably, MIF-KO CAC mice developed more severe disease compared with WT CAC mice. Furthermore, FMT experiments revealed that the fecal microbiota from MIF-KO donors was associated with increased tumor burden in WT recipients under CAC-inducing conditions compared with that in recipients colonized with WT-derived microbiota. Together, these findings suggest that MIF deficiency is associated with gut microbiota remodeling during CAC and support a potential relationship between the MIF-dependent host context, microbial composition and colorectal cancer severity. Full article

Persistent high-risk human papillomavirus (HPV) infection drives cervical cancer, a leading cause of cancer-related mortality among women in low- and middle-income countries; its clinical course is shaped by the cervicovaginal microbiome, in which Lactobacillus-dominated communities are associated with enhanced viral clearance. Despite this, vaginal probiotic interventions often demonstrate limited colonization efficiency, and autochthonous strain libraries from Central Asia remain absent. We applied a tiered functional screening workflow to a collection of 235 vaginal lactic acid bacterial isolates recovered from 400 women undergoing routine gynecological examination in Astana, Kazakhstan. The workflow sequentially filtered isolates on (i) antimicrobial activity against seven urogenital indicator pathogens using the deferred antagonism assay, (ii) surface adhesion by the Brilis erythrocyte assay, and (iii) biofilm-forming capacity by crystal violet retention and laser-capture-microdissection (LCM) microscopy. Species-level identification of the selected candidate was performed by whole-genome shotgun sequencing followed by Kraken2 taxonomic classification. From 235 isolates, three rounds of phenotypic filtering identified four broad-spectrum antimicrobial candidates (127-3, 127-4, 107-2, 107-4) with non-overlapping inhibitory profiles against seven urogenital indicator strains. Adhesion phenotyping segregated candidates into low- and moderate-adhesion groups, with none reaching the high-adhesion threshold. Among all four candidates, only strain 127-4 produced a reproducible biofilm-associated signal (crystal violet retention OD490 = 0.09 ± 0.07 at 24 h; 0.08 ± 0.03 at 48 h), consistent with early surface attachment under static conditions. Whole-genome shotgun sequencing assigned 97.81% of classified reads to Lactiplantibacillus plantarum, supporting preliminary identification of the selected isolate as L. plantarum strain 127-4. Composite ranking confirmed 127-4 as the only isolate combining broad antimicrobial activity (5/7 indicators), moderate adhesion (specific adhesion index, SPA = 2.95), and a detectable biofilm-associated phenotype. We report the first systematic functional screening of autochthonous cervicovaginal lactic acid bacteria from a Central Asian population and identify L. plantarum 127-4 as a probiotic candidate with an integrated trait profile rarely identified through single-criterion screening approaches. Beyond candidate identification, this work establishes a transferable workflow for assembling functionally annotated vaginal Lactobacillus collections from underrepresented populations, providing a foundation for future population-specific probiotic interventions targeting cervicovaginal health. Full article

Background/Objectives: Filipendula ulmaria (L.) Maxim. (meadowsweet) is a medicinal plant traditionally used for its antioxidant and anti-inflammatory effects. There is also some data indicating its anticancer potential; however, its impact on colorectal cancer cells remains poorly understood. Here we investigated the cytotoxic and pro-apoptotic effects of an aqueous F. ulmaria extract on human LoVo colorectal cancer cells and analyzed some of the mechanisms underlying it. Methods: LoVo colorectal cancer cells were treated with the aqueous extract and analyzed for intracellular reactive oxygen species (ROS), mitochondrial membrane potential, DNA damage, lysosomal alterations, apoptosis-related mechanisms, and antioxidant activity. Phytochemical profiling was performed by HPLC-TOF/MS. Results: The extract elevated intracellular ROS levels, disrupted mitochondrial membrane potential, and induced DNA damage in LoVo cells. Activation of crucial caspases, along with increased p53 levels, confirmed engagement of both extrinsic and intrinsic apoptotic pathways. Changes in lysosomal fluorescence were also observed, indicating alterations in lysosomal properties. In chemical assays (FRAP, TAC, DPPH, ABTS, and superoxide scavenging), the extract demonstrated robust antioxidant capacity comparable to or exceeding that of ascorbic acid. Phytochemical profiling by HPLC-TOF/MS revealed a rich presence of bioactive flavonoids, phenolic acids, and coumarins. Altogether, our findings indicate that the extract’s cytotoxicity against colon cancer cells arises from a multifaceted mechanism involving oxidative stress, organelle dysfunction, and apoptosis induction. Conclusions: These results highlight F. ulmaria aqueous extract as a promising candidate for colorectal cancer phytotherapy as a form of supportive treatment and warrant further preclinical validation. Full article

Dextran sodium sulfate (DSS) was used to induce ulcerative colitis in a porcine model to investigate the transcriptional responses in inflamed colonic tissue. Eleven pigs were divided into two groups, of which five pigs were administered an oral dose of DSS daily for five days, whereas six non-treated pigs served as a control group. Differences in transcript expression between treated and control pigs identified 425 down-regulated and 780 up-regulated mRNAs in response to DSS treatment. Fifty-nine differentially expressed miRNAs were also identified, comprising 20 up-regulated and 39 down-regulated miRNAs. The top enrichment KEGG pathways for the up-regulated genes were breast cancer (ssc05224), gastric cancer (ssc05226), focal adhesion (ssc04510), and the PI3K-Akt signaling pathway (ssc04151). The top gene ontology terms for the up-regulated genes were blood vessel development (GO:0001568), extracellular matrix and external encapsulating structure (GO:0031012). Protein–protein interaction network analysis identified three hub genes, including LOXL1, MFAP2, and FSTL3. Seventeen high-confidence miRNA-mRNA pairs were recognized, and two genes (CD101 and AVL9) have been confirmed as targets for ssc-miR-24-3p by dual-luciferase reporter assay. Our study provides a better understanding of the key roles of mRNAs and miRNAs in regulating DSS-induced colitis in pigs and defines sets of coding and non-coding RNA transcripts, which may serve as intervention targets or biomarkers for ulcerative colitis. Full article

Background/Objectives: Cadherin-17 (CDH17, LI-cadherin) is a non-classical cadherin with an atypical structure and unique functions. CDH17 expression is restricted to normal intestinal epithelium. Furthermore, CDH17 functions as an oncoprotein that promotes tumor migration and invasion in colorectal, gastric, and pancreatic cancers. Therefore, CDH17 is an important diagnostic marker and therapeutic target. The CDH17-directed strategies, including monoclonal antibodies (mAbs), bispecific Abs, antibody–drug conjugates (ADCs), and chimeric antigen receptor (CAR) T cells, have been evaluated in preclinical and clinical studies. Therefore, developing mAbs that specifically recognize cell surface-expressing CDH17 is essential for advancing both tumor diagnosis and therapy. Methods: Anti-human CDH17 mAbs (named Ca₁₇Mabs) were developed by immunizing a mouse with CDH17-overexpressed cells and a high-throughput screening using flow cytometry. Results: Among Ca₁₇Mabs, a clone, Ca₁₇Mab-5 (IgG₁, κ) specifically recognized CDH17-overexpressed Chinese hamster ovary-K1 (CHO/CDH17) cells with no detectable cross-reactivity to 21 other CDHs by flow cytometry. Ca₁₇Mab-5 also detected endogenous CDH17 in human colorectal cancer cell lines, COLO201 and COLO205. The apparent dissociation constant (K_D) values of Ca₁₇Mab-5 for CHO/CDH17 and COLO205 were estimated as 1.5 × 10⁻⁸ M and 1.3 × 10⁻⁸ M, respectively. Furthermore, Ca₁₇Mab-5 detected endogenous CDH17 by Western blotting. In immunohistochemistry, Ca₁₇Mab-5 exhibited clear membranous staining in normal colon epithelium, colorectal, gastric, and pancreatic cancers. Conclusions: Ca₁₇Mab-5 is a versatile tool for detecting CDH17 and has potential for tumor diagnosis. Full article

Mismatch repair–deficient (dMMR) or microsatellite instability-high (MSI-H) colorectal cancers exhibit high mutational burden and abundant neoantigen formation, generating a highly immunogenic tumour microenvironment characterised by dense lymphocytic infiltration and strong sensitivity to immune checkpoint inhibition. In metastatic colorectal cancer, PD-1 blockade produces durable responses almost exclusively in dMMR tumours, establishing mismatch repair status as a predictive biomarker for immunotherapy responsiveness. Recent studies extending immune checkpoint inhibitors (ICIs) into the neoadjuvant setting for localised dMMR colorectal cancer have produced major pathological response rates exceeding 90%, with pathological complete response (pCR) rates frequently surpassing 60%, challenging traditional oncologic staging frameworks, particularly with respect to lymph node assessment. Baseline clinical nodal staging in dMMR tumours is complicated by immune-mediated lymphadenopathy. Reactive lymphoid hyperplasia driven by tumour antigen exposure frequently produces enlarged lymph nodes that mimic metastatic disease on cross-sectional imaging. Following neoadjuvant immunotherapy, treatment-related immune activation may further increase nodal size or metabolic activity, while pathological examination often reveals sterilised nodes or immune infiltration without viable tumour. Consequently, conventional radiologic criteria for nodal metastasis demonstrate limited specificity in this context. The discordance between imaging findings and pathological outcomes raises important implications for staging accuracy, response assessment, and treatment planning. This review examines the biological basis of lymphatic involvement in dMMR colorectal cancer, evaluates the performance of current imaging modalities for nodal staging, and summarises emerging evidence from neoadjuvant immunotherapy trials. Particular emphasis is placed on the interpretation of lymph node findings in the era of immune checkpoint blockade and the implications for surgical decision-making, organ preservation, and future staging paradigms. Full article