OBJECTIVE:Irinotecan, a standard therapeutic agent for metastatic colorectal cancer (mCRC), often faces significant limitations due to drug resistance, with treatment failure observed in approximately 30%-50% of patients, leading to poor clinical outcomes. This study aims to systematically elucidate the molecular mechanisms underlying irinotecan resistance in colorectal cancer (CRC) by constructing patient-derived organoid (PDO) models combined with single-cell transcriptomics technology. METHODS:PDO models were successfully established from irinotecan-resistant and sensitive CRC patients. Single-cell RNA sequencing (scRNA-seq) was performed on the organoids, analyzing the transcriptomic heterogeneity of 12,360 high-quality cells. Gene Set Variation Analysis (GSVA), transcriptional regulatory networks, and cell communication networks were employed to dissect the resistance mechanisms. RESULTS:Drug sensitivity assays demonstrated that the IC50 value of irinotecan in CRC5 was significantly higher than that in CRC11, which was entirely consistent with their respective clinical phenotypes. Single-cell sequencing identified CRC5-specific drug-resistant cell clusters, Cluster 1 and Cluster 6. Cluster 1 (MARCKSL1+) was characterized by the activation of the Wnt signaling pathway and extracellular matrix (ECM) remodeling, which collectively contributed to the maintenance of stem cell-like properties, while Cluster 6 (AKR1C3+) exhibited significant enrichment in lipid metabolism and the Notch signaling pathway. CONCLUSION:This study integrates PDO models with single-cell transcriptomics technology to reveal key cell subpopulations and molecular mechanisms underlying irinotecan resistance. The core mechanisms driving resistance involve the activation of Wnt signaling and the synergistic effect of lipid metabolism-Notch pathways. Cluster 1 and Cluster 6 are identified as potential therapeutic targets, providing a theoretical basis for developing combination therapies targeting cancer stem cells or the metabolic microenvironment.

Colon cancer (CC), a significant global health burden with high incidence and mortality, is often accompanied by an immunosuppressive tumor microenvironment (TME) that limits the efficacy of immunotherapy. Immunoglobulin superfamily member 9 (IGSF9), a cell surface protein involved in cell adhesion and signaling, has been shown to promote tumor progression and regulate TME in other cancers, but its role in CC remains poorly understood. This study integrated multi-omics analyses, single-cell sequencing, and preclinical models to explore IGSF9's function in CC and its correlation with immunotherapy resistance. The results showed that IGSF9 was significantly upregulated in CC tumors, positively correlated with advanced tumor stages and poor prognosis such as reduced overall survival in colon adenocarcinoma (COAD). High IGSF9 expression was correlated with an immunosuppressive TME characterized by increased infiltration of regulatory T cells (Tregs), cancer-associated fibroblasts (CAFs), and reduced immune cell infiltration; it was also linked to lower tumor mutational burden (TMB) and microsatellite instability (MSI), predicting poor response to anti-PD-1 immunotherapy in clinical datasets. Single-cell and spatial transcriptomics revealed that IGSF9 was predominantly expressed in malignant epithelial cells, correlated with epithelial-mesenchymal transition (EMT) pathways. Drug sensitivity analysis identified Doramapimod, a MAPK inhibitor, which combined with anti-PD-1 therapy significantly enhanced tumor regression in mouse models by reducing Treg infiltration. In conclusion, this study establishes IGSF9 as a prognostic and predictive biomarker for immunotherapy resistance in CC and suggests that targeting IGSF9-associated KRAS/MAPK pathways with Doramapimod may offer a novel combination strategy to overcome TME-mediated resistance, warranting further clinical investigation for personalized CC treatment.

Chemotherapy resistance remains a major challenge in colorectal cancer (CRC) treatment, necessitating novel adjuvant strategies. This study employed an integrated analytical strategy combining network pharmacology, single-cell RNA sequencing (scRNA-seq) of patient-derived organoids (PDOs) and molecular dynamics simulations to identify bioactive compounds from Ganoderma lucidum and elucidate their chemo-sensitizing mechanisms. Network pharmacology identified five bioactive components of G. lucidum, corresponding to 267 potential targets. Integration with transcriptomic data, weighted gene co-expression network analysis (WGCNA), and known CRC genes refined these to 19 core targets. Cross-referencing with scRNA-seq data from irinotecan-treated PDOs pinpointed cyclin-dependent kinase 2 (CDK2) and Aurora kinase A (AURKA) as pivotal targets. Molecular dynamics simulations confirmed stable binding of the key component cerevisterol to both CDK2 and AURKA proteins, with binding free energies of -120.67 kJ/mol and -134.47 kJ/mol, respectively. In vitro cell viability assays across multiple CRC cell lines (HCT116, RKO, and HT-29) and PDOs demonstrated that cerevisterol significantly sensitized CRC cells to irinotecan (SN38). Notably, we observed that CDK2 was preferentially enriched in MSI-H tumors, whereas AURKA was enriched in MSS tumors, suggesting the potential of MSI status as a biomarker for patient stratification. Collectively, these findings identify cerevisterol as a dual-targeting natural product that modulates CDK2 and AURKA to overcome chemotherapy resistance, providing a quantitative analytical framework for discovering bioactive compounds and their molecular targets from medicinal fungi.

BackgroundNeoadjuvant PD-1/PD-L1 combined with chemoradiotherapy (CRT) has shown encouraging complete response (CR) rates in proficient mismatch repair (pMMR) or microsatellite-stable (MSS) locally advanced rectal cancer (LARC), yet immunotherapy duration varies widely across trials and its interaction with radiotherapy fractionation remains unclear.MethodsPubMed, Embase, and Cochrane Library were searched through October 8, 2025 for prospective trials of PD-1/PD-L1 inhibitors plus CRT in pMMR/MSS LARC. Random-effects models pooled CR (cCR+pCR) rates, with subgroup analyses and study-level meta-regression conducted to explore sources of heterogeneity. A linear probit model and an interactive quadratic probit model were applied to assess associations between treatment duration and response (PROSPERO number CRD420251253156).ResultsEighteen trials (972 patients; 10 LCRT-based and 8 SCRT-based) were included. Pooled CR and pCR rates were 44% (95% CI 0.38–0.49) and 40% (95% CI 0.36–0.45). CR was higher with SCRT plus ICI than LCRT plus ICI (50% vs 39%; P = 0.03), and pCR similarly favored SCRT (47% vs 34%; P < 0.01). Meta-regression identified radiotherapy fractionation and immunotherapy duration as study-level factors associated with CR. In analyses restricted to randomized controlled trials (RCTs), longer immunotherapy duration was positively associated with response. Exploratory quadratic models using all eligible studies suggested an inverted U-shaped pattern for LCRT (modeled peak at approximately 4.4 months) and a U-shaped pattern for SCRT (modeled trough at approximately 2.5 months).ConclusionAmong the included RCTs, longer immunotherapy duration was associated with higher response rates. The nonlinear patterns observed in the combined dataset were exploratory, may reflect heterogeneity between studies, and require prospective validation before informing treatment duration.Systematic review registrationhttps://www.crd.york.ac.uk/PROSPERO/view/CRD420261348026, identifier CRD420251253156.

Identifying reliable biomarkers to stratify colon cancer (CC) patients for immunotherapy remains a critical unmet clinical need due to the immunosuppressive nature of the tumor microenvironment. This study utilized TCGA-COAD transcriptomic data to classify patients into immunity-high and immunity-low phenotypes via single-sample GSEA, finding that the immunity-low subgroup exhibited significantly poorer survival (p = 0.038). Within this cohort, high expression of TNFRSF18 was identified as a key marker of inferior prognosis (p = 0.030). Single-cell RNA sequencing pinpointed TNFRSF18 expression primarily to Regulatory T cells (Tregs). Further validation in an independent immunotherapy cohort revealed that high infiltration of TNFRSF18+ Tregs significantly correlated with shorter disease-free and overall survival (p = 0.041 and p = 0.007). Functional ex vivo organoid experiments demonstrated that TNFRSF18-low tumors were susceptible to anti-PD-1 treatment, characterized by increased IFN-gamma and GZMB secretion, whereas TNFRSF18-high tumors displayed therapeutic resistance. In conclusion, this study establishes TNFRSF18+ Tregs as a novel prognostic marker and a predictor of immunotherapy response in immunosuppressed CC, suggesting that targeting TNFRSF18 could potentially enhance the efficacy of anti-PD-1 therapies in resistant patient subsets.

The Tianhe Procedure is a functional sphincter-preserving surgical approach developed for patients with rectal cancer following radiotherapy. This technique involves proximal extended resection of the colon beyond the pelvic cavity, followed by anastomosis of the non-irradiated proximal colon to the distal rectum or anal canal. This strategy aims to reduce the incidence of anastomotic complications and postoperative bowel dysfunction. However, there is currently a lack of standardized practice guidelines for implementing the Tianhe Procedure in China. Therefore, the Chinese Radiation Intestinal Injury Research Group, the Colorectal Surgery Group of Surgery Branch of the Chinese Medical Association, the Anorectal Branch of Chinese Medical Doctor Association, the Colorectal Cancer Committee of the Chinese Medical Doctor Association, the Colorectal Cancer Committee of China Anti-cancer Association, and the Gastrointestinal Surgical Branch of Guangdong Medical Doctor Association have jointly convened a panel of national experts to discuss and establish this standardized surgical procedure. This standard, based on the latest evidence from literature, research advancements, and expert experience, focuses on key aspects of the Tianhe Procedure, including its precise definition, indications, critical procedural steps, postoperative complications, and functional rehabilitation strategies. It aims to promote standardized implementation and broader clinical adoption of this innovative surgical technique.

Colorectal cancer (CRC) is a globally prevalent malignancy with rising incidence and mortality rates over the past decades. N6-methyladenosine (m6A) is the most abundant internal RNA modification in eukaryotes, and plays a pivotal role in post-transcriptional regulation. m6A is dynamically modulated by three core components, namely methyltransferases (writers), demethylases (erasers), and binding proteins (readers), which together govern the transcription, processing, translation, decay, and stability of mRNA. There has been accumulating evidence for the association of dysregulated m6A modification with CRC pathogenesis, metastasis, and therapeutic resistance. This review summarizes the biogenesis of m6A modification and its regulatory mechanisms, and discusses the dysregulation of m6A-related factors in CRC and the functional impacts. Most importantly, the review highlights the key roles of m6A modification in mediating CRC resistance to chemotherapy, targeted therapy, and immunotherapy. These insights may facilitate the development of novel therapeutic strategies for CRC.

We identified novel Molecular subtypes according to the expression of fibrosis-related genes (FRGs) and constructed a prognostic model using different expression genes (DEGs) for patients with Hepatocellular carcinoma (HCC). We downloaded the clinical data and transcriptome data of HCC from The Cancer Genome Atlas (TCGA) database, Gene Expression Omnibus (GEO) database, and International Cancer Genome Consortium (ICGC) database. We identified two fibrosis-related molecular subtypes of HCC by consensus unsupervised clustering analysis. Interestingly, these two molecular subtypes significantly differed in overall survival (OS) and clinical characteristics. Besides, the most minor absolute shrinkage and selection operator (Lasso) and multivariate Cox regression analysis were performed to develop a novel prognostic model by three genes (including KPNA2, LPCAT1, and AKR1D1). There was a statistically significant difference in OS between the high-risk and low-risk groups. The area under the ROC curve (AUC) of OS in 1-, 3-, and 5-year were satisfactory. Besides, the risk score was connected with critical clinical characteristics and could be an independent factor in predicting prognosis. Then, the nomogram was built by incorporating risk scores with clinical parameters. Additionally, the risk score was remarkedly correlated with TME and drug susceptibility. Finally, the results of H E staining and immunohistochemistry of Ki67 showed that the tumor of higher-risk patients are more malignant. The FRGs-based subtype and signature explain the HCC heterogeneity, which might provide a new method to develop a more efficient treatment.

Background Colorectal cancer (CRC) is a leading malignancy worldwide. Patient-derived organoids (PDOs) represent a cutting-edge model for cancer research. Atractylenolide I (ATT-1), a key bioactive lactone component of Baizhu (Atractylodes macrocephala), has demonstrated anti-cancer potential, but its mechanism of action in clinically relevant CRC PDO models remains unknown. Purpose To evaluate the anti-tumor efficacy and molecular mechanism of ATT-1 using colorectal cancer PDOs. Methods We investigated ATT-1 in CRC PDOs, assessing its effects on viability, proliferation (EdU), apoptosis (TUNEL), and DNA damage (γ-H2AX). Combination therapies with oxaliplatin or NU7441 were also evaluated in the PDO models. Multi-omics analyses (RNA-seq and metabolomics) profiled transcriptional and metabolic alterations. Direct target engagement was identified through LiP-MS and CETSA, and validated via shRNA knockdown. Molecular docking and dynamics simulations characterized the binding interactions. In vivo efficacy, safety, and combination therapy were validated in mouse xenograft models. Results ATT-1 selectively inhibited the proliferation of CRC PDOs, inducing apoptosis and inhibiting DNA damage repair. Multi-omics profiling revealed disruption of lipid metabolism, nucleotide synthesis, and cell cycle progression. We identified CLTC and XRCC5 as direct protein targets of ATT-1, validated by LiP-MS, CETSA, and functional knockdown assays. ATT-1 synergized with oxaliplatin and NU7441, enhancing anti-tumor efficacy in both PDO and xenograft models without significant hepatorenal toxicity. Furthermore, ATT-1 exhibited favorable pharmacokinetic properties. Conclusion This study identifies ATT-1 as a promising multi-targeted therapeutic for colorectal cancer by leveraging PDOs for direct, de novo target discovery. We uniquely identified two novel, high-affinity targets, CLTC and XRCC5, and elucidated a convergent dual-targeting mechanism wherein ATT-1 binding disrupts DNA damage repair and triggers apoptosis. This novel mechanism and its potent synergy with standard chemotherapy in physiologically relevant models provide a compelling strategy for integrating traditional Chinese medicine into modern precision oncology.

The aim of the present study was to investigate whether a multimodal radiomics model powered by machine learning could accurately predict the occurrence of metachronous liver metastasis (MLM) in patients with colorectal cancer (CRC). A total of 157 patients diagnosed with CRC between 2010 and 2020 were retrospectively included in the present study; of these patients, 67 patients developed liver metastases within 2 years of treatment, while the remaining patients (n=90) did not. Radiomics features were extracted from annotated MR images of the tumor and portal venous phase CT images of the liver in each patient. Subsequently, machine learning-based radiomics models were developed and integrated with the clinical features for MLM prediction, employing Least Absolute Shrinkage and Selection Operator and Random Forest algorithms. The performance of the models were evaluated using the receiver operating characteristic curve analysis, while the clinical utility was measured using the decision curve analysis. A total of 922 and 1,082 radiomics features were extracted from the MR and CT images of each patient, respectively, which quantified the intensity, shape, orientation and texture of the tumor and liver. The mean area under the curve (AUC) values for the prediction of MLM were 0.80, 0.68 and 0.82 for the CT, MRI and merged models, respectively. For the clinical and clinical-merged models, the AUC values were 0.62 and 0.75, respectively. There was no significant difference between the CT model and the merged model (P>0.05). In conclusion, the preliminary results of the present study demonstrated the utility of machine learning-based radiomics models in the prediction of MLM in patients with CRC. However, further research is warranted to explore the potential of multimodal fusion models, due to the minimal improvement observed in diagnostic performance.

BACKGROUND:Current evidence has shown an oncologic benefit of preoperative chemotherapy for patients with locally advanced and metastatic colon cancer. However, some patients present with stenosis that blocks the colonoscope but without symptomatic obstruction. Whether the stenosis will progress to obstruction during chemotherapy is unknown, and this uncertainty may encourage unnecessary diversion or stenting, or impede optimal treatment. METHODS:We retrospectively analyzed 465 consecutive patients with endoscopically stenotic colon cancer without symptomatic obstruction in a quaternary referral center from 2013 to 2020. Patients were grouped into upfront surgery (361) and preoperative chemotherapy (104). Clinical courses of patients receiving preoperative chemotherapy were described, with expedited surgery as the primary outcome. RESULTS:Obstruction or perforation occurred in 25 of 104 (24.0%) patients receiving preoperative chemotherapy, of which 11 discontinued chemotherapy and received expedited surgery, and 14 resumed chemotherapy after stent or medical treatment. In total, 93 (89.4%) patients received elective surgery after preoperative chemotherapy. In addition, the preoperative chemotherapy group demonstrated a 41.3% objective response rate. CONCLUSION:Three of 4 patients with stenotic colon cancer without symptomatic obstruction can complete preoperative chemotherapy uneventfully. Expedited surgery was needed in 10.6% because of obstruction or perforation during chemotherapy.

Importance Previous studies have demonstrated the advantages of short-term histopathological outcomes and complications associated with transanal total mesorectal excision (TME) compared with laparoscopic TME. However, the long-term oncological outcomes of transanal TME remain ambiguous. This study aims to compare 3-year disease-free survival of transanal TME with laparoscopic TME. Objective To evaluate 3-year disease-free survival between transanal TME and laparoscopic TME in patients with rectal cancer. Design, Setting, and Participants This randomized, open-label, noninferiority, phase 3 clinical trial was performed in 16 different centers in China. Between April 2016 and June 2021, a total of 1115 patients with clinical stage I to III mid-low rectal cancer were enrolled. The last date of participant follow-up was in June 2024. Interventions Participants were randomly assigned in a 1:1 ratio before their surgical procedure to undergo either transanal TME (n = 558) or laparoscopic TME (n = 557). Main Outcomes and Measures The primary end point was 3-year disease-free survival, with a noninferiority margin of -10% for the comparison between transanal TME and laparoscopic TME. Secondary outcomes included 3-year overall survival and 3-year local recurrence. Results In the primary analysis set, the median patient age was 60 years. A total of 692 male and 397 female patients were included in the analysis. Three-year disease-free survival was 82.1% (97.5% CI, 78.4%-85.8%) for the transanal TME group and 79.4% (97.5% CI, 75.6%-83.4%) for the laparoscopic TME group, with a difference of 2.7% (97.5% CI, -3.0% to 8.1%). The lower tail of a 2-tailed 97.5% CI for the group difference in 3-year disease-free survival was above the noninferiority margin of -10 percentage points. Furthermore, the 3-year local recurrence was 3.6% (95% CI, 2.0%-5.1%) for transanal TME and 4.4% (95% CI, 2.6%-6.1%) for laparoscopic TME. Three-year overall survival was 92.6% (95% CI, 90.4%-94.8%) for transanal TME and 90.7% (95% CI, 88.3%-93.2%) for laparoscopic TME. Conclusions and Relevance In patients with mid-low rectal cancer, 3-year disease-free survival for transanal TME was noninferior to that of laparoscopic TME.

Colorectal cancer (CRC) is characterized by high recurrence and metastasis, resulting in low patient survival. Dysbiosis of intestinal flora increases CRC metastasis risk. Mesalazine, an anti-inflammatory drug with proven antitumor activity, is commonly used to treat CRC in patients with inflammatory bowel disease, but whether it mediates CRC hepatic metastasis by affecting key flora is unknown. An animal model of CRC liver metastasis was established by injecting HCT116 cells into the spleens of mice and treated with mesalazine. Mouse feces were collected for 16S rRNA sequencing to analyze the abundance of intestinal flora. In vitro experiments demonstrated that mesalazine inhibited proliferation, migration, and invasion of CRC by modulating Bacteroides fragilis. Rescue experiments validated molecular mechanisms. Mesalazine inhibited CRC hepatic metastasis and reduced the abundance of Bacteroides fragilis in the feces of CRC hepatic metastatic mice by 16S rRNA sequencing analysis. In vitro experiments showed that mesalazine inhibited proliferation, migration, and invasion of CRC by affecting Bacteroides fragilis, mainly through the histone deacetylase 3 (HDAC3)/Wingless/Int/Beta-catenin (Wnt/β-catenin) axis. The study elucidated the mechanism by which mesalazine inhibited CRC liver metastasis through the HDAC3/Wnt/β-catenin axis by affecting the abundance of Bacteroides fragilis, which provided a new idea for CRC treatment.

BACKGROUND:Gastric cancer is a biologically aggressive disease, accounting for a substantial proportion of cancer-related deaths globally. Accurate localization of the lesion by artificial intelligence techniques helps timely and efficiently diagnose and treat. Segment Anything Model (SAM) has demonstrated considerable potential in medical image segmentation by displaying high performance in numerous image benchmark tests. However, its resource-intensive nature limits feasibility in embedded medical contexts. METHODS:This study proposed GC-SAM, a lightweight model for tumor segmentation. The architecture of GC-SAM is innovatively proposed, including a knowledge distillation image encoder, prompt encoder, and mask decoder, which effectively replaces the conventional fixed and computationally intensive network components. RESULTS:Extensive experiments demonstrate that GC-SAM significantly outperforms both classical segmentation models and recent state-of-the-art networks. On the internal test set, GC-SAM achieves 0.8186 Dice and 0.6504 mIoU, while reducing inference time and parameter count by over 80% compared to the original SAM. On the external dataset, GC-SAM maintains superior performance (Dice 0.8350), demonstrating excellent generalization. CONCLUSIONS:The proposed GC-SAM model shows strong capability in segmenting gastric cancer tissue, while also demonstrating practical potential for deployment in embedded medical imaging devices.

Immunotherapy has shown limited efficacy in colorectal cancer (CRC), necessitating the urgent identification of novel immunotherapeutic targets to address this challenge. Tertiary lymphoid structures (TLSs) are complex immune structures that spontaneously form within and outside of the tumor microenvironment, resembling secondary lymphoid organs. The formation of TLS formation dynamically adapts to the tumor microenvironment, with their immune function maintained by resident immune cells and cytokine networks. TLS develops antitumor immune competence following dynamic reorganization of their cellular components and proportions, although the precise mechanistic underpinnings remain to be fully characterized. Numerous studies have demonstrated that TLS can modulate the immune status of CRC, potentially enhancing the efficacy of immunotherapy. Currently, the clinical setting lacks appropriate immunological markers to assess immunotherapy outcomes, and TLS may offer a solution to this limitation. This article will review current research on TLS as a novel therapeutic target in the immunotherapy of malignant colorectal tumors.

Objective: To explore the diagnostic value of combined detection of bladder tumor antigen (BTA), bladder tumor antigen-associated test (BTA stat), nuclear matrix protein 22 (NMP22), and survivin in urothelial carcinoma. Methods: Sixty patients with urothelial carcinoma admitted from January 2024 to January 2025 were selected as the observation group for this study, and 60 healthy individuals were selected as the control group. BTA, BTA stat, NMP22, and survivin tests were performed on both groups, respectively. The test results were analyzed to evaluate the diagnostic value of combined detection. Results: The levels of BTA, BTA stat, NMP22, and survivin in the observation group were higher than those in the control group (P < 0.05). The specificity, sensitivity, and accuracy of combined detection of BTA, BTA stat, NMP22, and survivin were higher than those of single detection methods (P < 0.05). There were significant differences in the positive rates of BTA, BTA stat, NMP22, and survivin among patients with different tumor diameters, tumor numbers, pathological grades, clinical stages, and lymph node metastasis status (P < 0.05). Conclusion: In the diagnosis of urothelial carcinoma, the combined detection of BTA, BTA stat, NMP22, and survivin has high diagnostic value and can be promoted and applied in clinical diagnosis.

OBJECTIVE:To evaluate the diagnostic performance of multiple photon-counting CT (PCCT)-derived parameters for hepatic fat quantification, using MRI-derived proton density fat fraction (PDFF) as the reference standard. METHODS:This prospective study included 59 adult participants who underwent both MRI and PCCT within 2 h. Standardized CT values, Liver-virtual non-contrast (VNC) fat fraction, and liver-to-spleen/fat-derived indices (L-S, L/S, L-F, L + F, L/F) were obtained from non-contrast PCCT. MRI PDFF was derived using a 6-echo q-Dixon sequence. Pearson and Spearman correlation analyses were used to assess the association between PCCT-derived parameters and MRI PDFF. Linear mixed-effects model (LMM) was performed to establish a conversion formula between CT attenuation and MRI PDFF. RESULTS:All PCCT-derived parameters were significantly correlated with MRI PDFF. Standardized CT values demonstrated the strongest correlation (r = -0.94, p < 0.001), followed by L-S (r = -0.90, p < 0.001), L/F (r = 0.89, p < 0.001), and L/S (r = -0.88, p < 0.001). Liver-VNC fat fraction showed a relatively lower correlation (r = 0.84, p < 0.001). LMM revealed a significant association between standardized CT values and MRI PDFF (β = -0.55, p < 0.001), with the regression equation: MRI PDFF (%) = -0.55 × CT (HU) + 41. CONCLUSION:Among various attenuation-based PCCT parameters, the standardized CT value shows the strongest correlation with MRI PDFF and is clinically easy to obtain. It may serve as a promising noninvasive alternative to MRI for hepatic fat quantification. However, due to the limited sample size, these findings should be interpreted with caution.

OBJECTIVE:The objective of this study is to examine the impact of KW2478 combined with DDP on colorectal cancer cells both in vitro and in vivo and to elucidate the molecular mechanism of KW2478 in colorectal cancer. METHODS:qRT-PCR and Western blot were employed to assess HSP90 mRNA and protein expression in normal intestinal epithelial and colorectal cancer cells. DLD-1 and HCT116 were selected for the experiment. CCK-8 was used to detect cytotoxicity; apoptosis rate was measured using flow cytometry; Western blot was employed to measure the expression levels of apoptotic and PI3K/AKT/mTOR pathway proteins. HCT116 was used to construct a subcutaneous tumor model in nude mice. After treatment with KW2478 and DDP, the growth rate, volume, and weight of the tumor were observed. The expression of Ki67 was detected by immunohistochemistry. Apoptosis of tumor cells was detected using TUNEL. Western blot was employed to measure the expression levels of apoptotic and PI3K/AKT/mTOR pathway proteins. RESULTS:HSP90 mRNA and protein levels were elevated in colorectal cancer cells compared to normal colorectal epithelial cells. HSP90 mRNA and protein expression levels were also significantly elevated in HCT116 and DLD-1 cells compared to other colorectal cancer cells. In DLD-1 and HCT116 cells, KW2478 and DDP inhibited cell viability. The combination of KW2478 and DDP exhibited a significantly higher inhibitory effect compared to either KW2478 or DDP alone. DDP markedly triggered apoptosis in HCT116 and DLD-1. KW2478 at 3 μg/ml and 6 μg/ml induced apoptosis in HCT116 cells but not in DLD-1 cells. The combination of KW2478 and DDP induced a significantly higher apoptosis rate as compared to either KW2478 or DDP alone. Treatment of HCT116 and DLD-1 with KW2478 or DDP alone increased Bax, Caspase9, and Caspase3 protein expression, and decreased BCL-2. The KW2478 + DDP combined treatment group exhibited more significant changes. Phosphorylation of PI3k, AKT, and mTOR decreased in the KW2478 or DDP treatment groups, with more significant changes observed in the KW2478 + DDP combination group. The growth rate, volume, and weight of subcutaneous tumors in the KW2478 or DDP treatment groups were significantly lower than control, and the KW2478 + DDP combination group was more affected. Ki67 expression in subcutaneous tumors was reduced in the KW2478 or DDP treatment groups compared to the vehicle control group, with the lowest expression observed in the KW2478 + DDP combination group. The fluorescence intensity of subcutaneous tumors was higher in both the KW2478 and DDP treatment groups compared to the vehicle control group, and the KW2478 + DDP combination group exhibited the strongest fluorescence intensity among them. CONCLUSION:The combination of KW2478 and cisplatin inhibits colorectal cancer cell proliferation and induces apoptosis by regulating the PI3K/AKT/mTOR pathway.

The National Health Commission of the People's Republic of China Guidelines for Diagnosis and Treatment of Colorectal Cancer (2025 edition), based on evidence-based medicine, integrates cutting-edge international advances with Chinese clinical practice, and supplements and completes the previous versions. This version of the guidelines, retains the core diagnostic and treatment framework, highlights new contents such as "Surgical treatment of anal canal cancer" and "New technologies and advances in diagnosis and treatment", and systematically summarizes the core points in the surgical treatment, medical oncology treatment, radiation oncology treatment, imaging, and pathology treatment. It is designed to help clinicians quickly grasp the key points of the guidelines and promote the standardization, precision, and consistence of colorectal cancer diagnosis and treatment.