Metastatic Colon Cancer Research Articles

Preoperative CT lymph node size as a predictor of nodal metastasis in resectable Colon cancer: a retrospective study of 694 patients

PurposeThis study aimed to investigate the efficacy of measuring lymph node size on preoperative CT imaging to predict pathological lymph node metastasis in patients with colon cancer to enhance diagnostic accuracy and improve treatment planning by establishing more reliable assessment methods for lymph node metastasis.MethodsWe retrospectively analyzed 1,056 patients who underwent colorectal resection at our institution between January 2004 and March 2020. From this cohort, 694 patients with resectable colon cancer were included in the study. We analyzed the relationship between lymph node size on preoperative CT imaging and lymph node metastasis identified on postoperative pathological examination.ResultsThe optimal cutoff values for the maximum long diameter and short diameter of regional lymph nodes on preoperative CT were identified as 6.5 mm and 5.5 mm, respectively, with an AUC of 0.7794 and 0.7755, respectively. Notably, the predictive accuracy varied by tumor location. Higher cutoff values were observed in the right-sided colon (maximum long diameter: 7.7 mm, maximum short diameter: 5.9 mm) compared to the left-sided colon (maximum long diameter: 5.8 mm, maximum short diameter: 5.2 mm).ConclusionLymph node size on preoperative CT is a significant predictor of pathological lymph node metastasis in colon cancer. Notably, the optimal cutoff values for predicting lymph node metastasis vary depending on the specific region within the colon.

An artesunate-modified half-sandwich iridium(iii) complex inhibits colon cancer cell proliferation and metastasis through the STAT3 pathway.

Colon cancer is one of the most commonly diagnosed cancers and is recognized as the most aggressive tumor of the digestive system. Aberrant activation of signal transducer and activator of transcription 3 (STAT3) is associated with proliferation, metastasis and immunosuppression of the tumor cells. Here, to inhibit the STAT3 pathway and suppress metastasis in colon cancer cells, the half-sandwich iridium complex Ir-ART containing an artesunate-derived ligand was synthesized. The complex showed remarkable antiproliferative activity against human colon cancer HCT-116 cells and exhibited a concentration-dependent reduction in STAT3 protein expression. Mechanism study demonstrates that Ir-ART is located mainly in the nucleus and mitochondria, causing γ-H2AX and cyclin B1 reduction and reactive oxygen species accumulation and mitochondrial membrane potential loss, ultimately leading to autophagic cell death. The migration of cancer cells was also inhibited via metalloproteinase 9 downregulation. Furthermore, Ir-ART could initiate antitumor immune responses by eliciting immunogenic cell death and downregulating immunosuppressive cytokine cyclooxygenase-2. Taken together, Ir-ART is expected to be further applied to chemotherapy and immunotherapy for colon cancer.

Unraveling the mechanism of microRNA-134 in colon cancer progression: Targeting KRAS and PIK3CA for cell cycle control and histone deacetylase regulation.

Colon cancer is the leading cause of cancer-related deaths worldwide. MicroRNAs (miRNAs) are key regulators of gene expression, often dysregulated in colon cancer. This study aims to elucidate the therapeutic role of miR-134-5p as a tumor suppressor miRNA in colon cancer cells. We analyzed miRNA expression profiles in primary and metastatic colon cancer cells. The clinical significance of miR-134-5p was evaluated using the TCGA database. Bioinformatics tools (HADDOCK) predicted miRNA-mRNA interactions and the molecular docking of miRNA-mRNA-AGO2 complexes. Luciferase reporter assays, cell proliferation, immunofluorescence, colony forming unit assays, and qRT-PCR analysis assessed miR-134-5p effects on KRAS, PIK3CA, and downstream signaling pathways in primary and metastatic colon cancer cells. miR-134-5p was downregulated in colon cancer cells. Bioinformatics analysis suggested KRAS, PIK3CA, EGFR, and HDAC5 as potential targets. HADDOCK analysis revealed strong binding affinity and structural stability between KRAS, PIK3CA, miR-134-5p, and AGO2. Gene-reporter assays confirmed miR-134-5p-mediated degradation of KRAS and PIK3CA. miR-134-5p transfection reduced KRAS and PI3K protein levels, suppressed EGFR/RTK signaling and its downstream targets, and inhibited HDAC expression, ultimately reducing colon cancer cell proliferation. The results of this study confirm that miR-134-5p acts as a potential tumor suppressor miRNA in colon cancer cells by inhibiting KRAS and PI3K expression through AGO2-mediated gene silencing. It deregulates downstream EGFR signaling and HDACs, thereby reducing colon cancer cell proliferation. These findings highlight miR-134-5p as a promising therapeutic target for miRNA-mediated anticancer therapy.

Alternating Oxaliplatin and Irinotecan Chemotherapy Combined With Capecitabine and Bevacizumab for Microsatellite-stable Stage IV Metastatic Colon Cancer With a BRAF V600E Mutation: Two Case Reports Indicating Survival Improvement over Standard Therapy.

Colorectal cancer (CRC) has the third-highest incidence among human cancers. Advancements in chemotherapy and targeted therapy have improved the treatment outcomes for patients with CRC. However, the management of patients with unresectable metastatic CRC (mCRC) continues to be a significant challenge for clinicians worldwide, particularly for those with microsatellite stability (MSS) and the BRAF V600E mutation, as they are associated with the poorest prognosis. The present study describes two patients with unresectable MSS, BRAF V600E-mutated stage IV metastatic CRC using a biweekly alternating regimen of irinotecan and oxaliplatin combined with capecitabine and bevacizumab. Case 1 stabilized after alternating treatment, whereas Case 2 progressed after alternating treatment, with progression-free survival (PFS) of 20+ and 24.5 months, respectively. Circulating levels of carcinoemryonic antigen (CEA) dropped to near normal in both cases. A partial response (PR) was determined for both cases. The two cases suggest that an alternating chemotherapy regimen of oxaliplatin and irinotecan, combined with capecitabine and bevacizumab is effective in the treatment of MSS, BRAF V600E-mutated stage IV metastatic CRC. The progression-free survival was significantly prolonged (both exceeding 20 months) compared to the first-line standard chemotherapy regimen for this disease. With a good balance between toxicity and efficacy, this alternating chemotherapy regimen can be considered as a potential first-line option for microsatellite-stable metastatic colon cancer.

Synthesis and Characterization of 5-Fluoroacil Loaded Calcium Carbonate Nanoparticles for Targeted Cancer Therapy

Chemotherapy is often limited by its systemic toxicity and lack of specificity, necessitating the development of targeted drug delivery systems that can enhance therapeutic efficacy while minimizing adverse effects. Addressing this, our study aimed to synthesize and characterize 5-Fluorouracil loaded calcium carbonate nanoparticles derived from cockle shells. The research aimed at increasing site-specific drug release and reducing cytotoxicity. The nanoparticles were prepared using a simple co-precipitation method, ensuring eco-friendliness and cost-effectiveness. The encapsulation of 5-FU was confirmed by transmission electron microscopy (TEM), which revealed an increase in nanoparticle size from 19.2 ± 2.284 nm for the unloaded ones to 34.8 ± 4.066 nm for the 5-FU-loaded CaCO3 NPs. In vitro release studies demonstrated a pH-sensitive release profile, with more rapid drug release at pH 4.8 compared to pH 7.3. Biocompatibility assays on the HS-27 human skin fibroblast cell line indicated high cell viability, with over 90% maintained even at high nanoparticle concentrations (1000 µg/mL). In addition, cytotoxicity assays on the SW480 (primary colon cancer) and SW620 (metastatic colon cancer) cell lines showed a dose-dependent decrease in cell viability and demonstrated a more controlled and sustained release compared to free 5-FU, resulting in higher cell viability at all time points and concentra-tions. The 5-FU-loaded CaCO3 NPs significantly reduced the immediate cytotoxic effects observed with free 5-FU while effectively targeting cancer cells. These findings suggest that the 5-FU-CaCO3 NPs offer a promising alternative to conventional chemotherapy, providing targeted drug delivery with the potential for reduced systemic toxicity and enhanced therapeutic efficacy.

Investigation of extracellular metabolites in cancer and healthy colon cells in a time-dependent manner

Metabolite analysis is critical in the cancer field because of provides information about the metabolic status of cells. The profiling of extracellular metabolites presents technical advantages over intracellular metabolites, such as easier access to extracellular metabolites without a quenching method and the growth medium containing high biomass. This study aimed to investigate the extracellular level of metabolites in colon cancer cells in a time-dependent manner. One million cells were seeded in 4 Petri dishes and glucose, pyruvate, TCA cycle metabolites, and D2-HG in the conditioned medium were determined by DNS and HPLC method and pyruvate assay for 24-96th hours. The results showed that glucose is consumed, and pyruvate and TCA cycle intermediates are released in decreasing amounts in all cell lines. It was also observed that glucose was more consumed, and TCA cycle metabolites were less released in metastatic colon cancer cells (SW620) than in primary colon adenocarcinoma cells (Caco-2). Most importantly, D2-HG oncometabolite was released more into the growth medium of colon cancer cells than normal colon cells for four days. In conclusion, the D2-HG is highly produced and released to the growth medium of colon cancer cell lines in a cancer-type-specific manner.

High tumor mutational burden assessed through next-generation sequencing predicts favorable survival in microsatellite stable metastatic colon cancer patients

BackgroundMicrosatellite instability (MSI) is a well-established predictive biomarker for immune checkpoint inhibitor (ICI) response in metastatic colon cancer. Both high MSI and tumor mutational burden (TMB) are markers of genomic instability. However, the prognostic and predictive value of TMB in patients with microsatellite stable (MSS) tumors remains unclear.MethodsWe evaluated the prognostic significance of TMB levels in MSS metastatic colon cancer patients undergoing standard treatments. Tumor responses were assessed using RECIST v1.1 criteria. Comprehensive clinical and molecular profiling was conducted, including next-generation sequencing (NGS) for TMB evaluation with the TruSight Oncology® kit. Overall survival (OS) was the primary endpoint. Multivariate Cox regression analysis was utilized to assess the relationship among potential prognostic factors.ResultsAmong 102 MSS metastatic colon cancer patients, high TMB (> 10 mut/mb) was associated with a significantly longer median OS compared to low TMB (70.0 vs 45.0 months, respectively; HR: 0.45; 95% CIs 0.21 to 0.96; P = 0.0396). Multivariate analysis, adjusting for age, gender, number of metastatic sites, response to first-line chemotherapy, RAS mutational status, and liver involvement, identified TMB as an independent prognostic factor, along with response to first-line chemotherapy.ConclusionsOur results highlight the prognostic significance of TMB in MSS metastatic colon cancer patients, suggesting its potential role in patient stratification and treatment decision-making.

Expression and clinical significance of Numb and Notch-1 proteins between tissue of colon cancer and regional lymph node metastases.

To investigate the expression and clinical significance of Notch-1 and Numb protein in colon cancer tissues and regional lymph node metastases. Immunohistochemical method was used to detect the expression of Notch-1 protein and Numb protein in 110 cases of colon cancer tissues, along with tumor adjacent tissues and 56 cases of MLN tissues, and to analyze its role in colon cancer and MLN tissue. Comparing colon cancer tissue or lymph node metastases with tumor adjacent tissue, the positive expression rate of Numb was significantly decreased, while the positive expression of Notch-1 was significantly increased in colon cancer tissue or lymph node metastases (both p<0.05). The expression of Notch-1 and Numb was correlated with the lymph node metastasis, TNM stage, and degree of differentiation (p<0.05). The expression between Numb and Notch-1 showed negative correlation in colon cancer tissues (r=-0.261, p<0.05). There was no relationship between the expression of Numb and Notch-1 protein in colon cancer and metastatic lymph node tissue (p>0.05). Numb expression is decreased and Notch-1 expression is increased in colon cancer tissue and metastatic lymph node tissue, suggesting that the interaction between the two proteins may play a promote role in the development, invasion, and metastasis of colon cancer. There was no relationship between the expression of Numb and Notch-1 protein in colon cancer and metastatic lymph node tissue, suggesting that there is no obvious enhancement of the cancer cells; in the process of lymph node metastasis, the degree of malignant biological behavior remains relatively stable.

Prognostic and Molecular Characterization of Metastatic Transverse Colon Cancer: Insights From a Single-Center Retrospective Study.

Metastatic transverse colon cancer (TCC) represents a unique subset of colorectal cancer with features of both right and left colon tumors due to its distinct embryologic origin. This study retrospectively analyzes the clinical, pathological, and molecular factors influencing survival outcomes in TCC patients treated at a single center. For this study, we reviewed the files of 372 metastatic patients and analyzed the data of 71 patients with a diagnosis of TCC in detail. The remaining patients were patients with right or left colon tumors and we compared the overall survival (OS), molecular mutations (KRAS, NRAS, BRAF, MSI status), and clinicopathological features of our patients with transverse colon tumors with these patients. The median OS for TCC patients was 19.7 months, with metastasectomy and Eastern Cooperative Oncology Group (ECOG) performance status emerging as significant prognostic factors. Molecular analyses revealed KRAS mutations in 49% and BRAF mutations in 13% of TCC cases, aligning TCC closer to right-sided tumors in certain molecular characteristics. However, histopathologic diversity, including mucinous histology in 20% of TCC cases, indicated a need to consider TCC as a distinct entity. These findings underscore the complex biological nature of TCC and the necessity for tailored therapeutic approaches, especially as survival rates remain suboptimal. Further multicenter, prospective studies are recommended to establish refined treatment strategies for TCC patients.

Mismatch repair deficiency confers worse survival in stage IV colon cancer.

Metastatic colon cancer (MCC) is a debilitating condition with a poor prognosis. Currently, there is limited data that investigates MCC in relation to mismatch repair (MMR) status. The aims of this study are to compare sociodemographic and clinicopathologic features and mortality between patients with MMR-proficient (MMR-P) and MMR-deficient (MMR-D) MCC. We performed an 8-year retrospective review of the National Cancer Database (NCDB) to identify patients age ≥18 years with MCC and reported MMR status. Data collection included sociodemographic characteristics, primary tumor sites and histopathologic features, and treatment modalities. Outcomes included 90-day, 180-day, 1-year, and 2-year overall mortality. Bivariate logistic regression and multivariate Cox regression identified differences between MMR-P and MMR-D and identified predictors of mortality, respectively. A total of 10,922 MCC cases were identified; 8,796 (80.53%) were MMR-P and 2,126 (19.47%) were MMR-D. MMR-D was independently associated with older age at diagnosis, female sex, mucinous adenocarcinoma, medullary carcinoma, and lymph-vascular invasion. MMR-P was independently associated with perineural invasion and left-sided colonic primary tumor predominance. When adjusted for demographics, histology, and treatment modalities, MMR-D was associated with mortality at 180 days, 1 year, and 2 years. Our study identified several key sociodemographic and clinicopathologic features of MMR-D MCC. MMR-D appears to confer increased overall mortality at 180 days, 1 year, and 2 years after diagnosis in MCC.

Bevacizumab versus aflibercept with FOLFIRI after FOLFOX and bevacizumab in RAS mutant metastatic colon cancer a Turkish oncology group study

We aimed to compare FOLFIRI and bevacizumab with FOLFIRI and aflibercept in terms of overall survival (OS), progression-free survival (PFS) and safety in patients with RAS-mutant metastatic colon cancer who progressed after first-line FOLFOX or XELOX and bevacizumab treatment. This retrospective study included 243 patients from 15 different centres in Turkey. The endpoints of the study were OS, PFS and safety and side effect outcomes. The median age of the patients included in the study was 60 (21–85) years. Of the patients enrolled in the study, 114 patients (46.9%) received aflibercept and 129 patients (53.1%) received bevacizumab. Median OS was 11.2 (95% CI: 9.1–13.2) months in patients receiving FOLFIRI + aflibercept and 14.1 (95% CI: 11.2–17.1) months in patients receiving FOLFIRI + bevacizumab. The median PFS was 5.7 (95% CI: 4.9–6.5) months in the aflibercept arm and 7.7 (95% CI: 7.1–8.3) months in the bevacizumab arm. Grade 3–4 side effects were observed in 58 (50.9%) patients in the aflibercept arm and 33 (25.6%) patients in the bevacizumab arm. As a result of our study, in patients with metastatic RAS-mutant colon cancer who progressed after first-line oxaliplatin-based doublet chemotherapy and bevacizumab, better OS and PFS results were obtained in patients receiving bevacizumab with FOLFIRI compared to patients receiving aflibercept with FOLFIRI. In addition, the side effect profile was more tolerable in the bevacizumab arm.

Prognostic Immunological Factors of Lymphogenic Metastasis in Colon Cancer

Введение. Корректная оценка стадии и прогноза является основой успешного лечения любой опухоли. Цель — оценить роль ряда иммунологических параметров локального иммунитета при раке ободочной кишки (РОК) в зависимости от распространенности процесса и на основании полученных данных разработать комплексную модель для определения вероятности лимфогенного метастазирования опухоли. Материалы и методы. В работу включено 50 пациентов РОК: c поражением лимфатических узлов (N+) — 27 пациентов (54 %), без поражения (N0) — 23 пациента (46 %). Из ткани опухоли, перитуморальной зоны (1–3 см от опухоли), линии резекции (~10 см от опухоли) была получена клеточная суспензия для выявления основных субпопуляций лимфоцитов, а также определения экспрессии TLRs (2, 3, 4) на CD45+(лимфоцитах), CD45-EpCAM+ (эпителиальные клетки). Статистический анализ результатов исследования проводился с помощью программы STATISTICA 13.3. Результаты. В тканях опухоли у пациентов с N0 увеличено количество T-лимфоцитов, натуральных киллеров (NK). В перитуморальной зоне при N0 отмечено более низкое содержание CD19+, а при N+ — двойных позитивных лимфоцитов (ДП), NK (p &lt; 0,05). Для тканей опухолей группы N0, в отличие от N+, характерно уменьшение количества опухолевых клеток, экспрессирующих TLR3. В перитуморальной зоне обеих групп отмечено разнонаправленное изменение количества клеток, которые экспрессируют TLR2: уменьшение — при N0, увеличение — при N+(p &lt; 0,05). При анализе относительного количества лимфоцитов во фрагментах тканей первичных опухолей пациентов с N0 отмечено уменьшение относительного количества клеток, экспрессирующих TLR2, и увеличение экспрессирующих TLR3,4, а для пациентов N+ отмечено увеличение количества клеток, которые экспрессируют TLR4 в 2,5 раз (p &lt; 0,05). С помощью метода логистической регрессии была разработана комплексная модель для определения вероятности лимфогенного метастазирования РОК. Индивидуальные значения пациентов подставляются в математическую модель и рассчитывается вероятность N+ (диагностическая чувствительность — 89,1 %, специфичность — 88,2 %). Выводы. Предложенная модель позволит прогнозировать распространенность процесса на предоперационном этапе у больных РОК, а также может стать одним из методов уточняющей диагностики в стадировании процесса.

Pneumatosis intestinalis in metastatic colon cancer during cetuximab treatment

Pneumatosis intestinalis in metastatic colon cancer during cetuximab treatment

Metastatic colon cancer: clinical practice guideline update in 2024

This article presents changes to clinical guidelines for the treatment of metastatic colon cancer in 2024. The new provisions in the clinical guidelines are complemented by a brief overview of the research results that underlie them. The changes considered concern not only systemic antitumor treatment, but also surgery and molecular genetic diagnostics. The differences between the recommendations of RUSSCO and the Ministry of Health of Russia are given. The introduction of information to determine the clinical benefit of expensive therapeutic options in relation to the use of the ESMO-MCBS and RUSSCO-MCBS scales is discussed.Aim. Bringing information to a wide range of readers on planned changes in clinical guidelines.

Suppression of Metastasis of Colon Cancer to Liver in Mouse Models by Pretreatment with Extracellular Vesicles Derived from Nanog-Overexpressing Colon-26 Cancer Cells.

It has been demonstrated that cancer cells that have survived cancer treatment may be more malignant than the original cancer cells. These cells are considered the main cause of metastasis in prognosis. A Nanog-overexpressing colon-26 (Nanog+colon26) was generated to obtain such a malignant cancer cell model, which was confirmed by enhancement of metastatic potential by in vivo tests using mice. Extracellular vesicles (EVs) secreted from Nanog+colon26 cells (Nanog+colon26EVs) were administered to mice three times per week for three weeks. Subsequently, Nanog+colon26 cells were administered, and metastatic colonies were analyzed two weeks later. The results demonstrated that the administration of EVs suppressed metastasis. Nanog+colon26EVs enhanced phagocytic activity and M1 marker CD80 of a macrophage cell line J774.1. These suggested the enforcement of tumor-suppressive properties of macrophages and their contribution to the in vivo suppression of metastasis. Small RNA sequencing was conducted to identify Nanog-dependent miRNAs that exhibited significant changes (Fc ≥ 1.5 or Fc ≤ 1/1.5; p < 0.05) in Nanog+colon26EVs relative to colon26EVs. Nine miRNAs (up-regulated: four, down-regulated: five) were identified, and 623 genes were predicted to be their target genes. Of the 623 genes identified, nine genes were predicted to be highly relevant to macrophage functions such as phagocytosis.

Spatially Resolved in vivo CRISPR Screen Sequencing via Perturb-DBiT.

Perturb-seq enabled the profiling of transcriptional effects of genetic perturbations in single cells but lacks the ability to examine the impact on tissue environments. We present Perturb-DBiT for simultaneous co-sequencing of spatial transcriptome and guide RNAs (gRNAs) on the same tissue section for in vivo CRISPR screen with genome-scale gRNA libraries, offering a comprehensive understanding of how genetic modifications affect cellular behavior and tissue architecture. This platform supports a variety of delivery vectors, gRNA library sizes, and tissue preparations, along with two distinct gRNA capture methods, making it adaptable to a wide range of experimental setups. In applying Perturb-DBiT, we conducted un-biased knockouts of tens of genes or at genome-wide scale across three cancer models. We mapped all gRNAs in individual colonies and corresponding transcriptomes in a human cancer metastatic colonization model, revealing clonal dynamics and cooperation. We also examined the effect of genetic perturbation on the tumor immune microenvironment in an immune-competent syngeneic model, uncovering differential and synergistic perturbations in promoting immune infiltration or suppression in tumors. Perturb-DBiT allows for simultaneously evaluating the impact of each knockout on tumor initiation, development, metastasis, histopathology, and immune landscape. Ultimately, it not only broadens the scope of genetic inquiry, but also lays the groundwork for developing targeted therapeutic strategies.

Main aspects of personalized approach to the treatment of patients with chemotherapy resistant metastatic colorectal cancer

Purpose of the study. To improve the results of treatment of patients with unresectable metastases of colorectal cancer in the liver that are not controlled by systemic chemotherapy.Materials and methods. The study includes clinical data on the treatment of 76 patients with metachronous metastases of colorectal cancer in the liver that are not controlled by systemic chemotherapy. Patients underwent removal of the primary tumor according to urgent indications at the first stage of complex treatment, followed by systemic chemotherapy in an adjuvant mode. After 24.5 ± 0.2 months, patients were diagnosed with metastatic liver damage, and therefore systemic chemotherapy was initiated. After changing two lines of drug therapy with a registered progression of the oncological process, liver metastases were recognized as uncontrolled by systemic chemotherapy. After that patients were included in the given study and divided into two groups. The study group included 40 patients who underwent regional chemotherapy. The control group included 36 patients who continued systemic chemotherapy with subsequent line changes. The effectiveness was evaluated according to the RECIST 1.1 and mRECIST scales, as well as the overall one-year, two- and three-year survival rates.Results. The median overall survival of patients in the control and study groups was 30.0 ± 0.8 and 41.5 ± 0.5 months, respectively, p &lt; 0.05. The total one-year, two- and three-year survival of patients in the control and study groups was 94.4 %, 69.4 %, 33.3 % and 100 %, 82.5 %, 57.5 %, respectively, p &lt; 0.05. The median life expectancy of deceased patients in the control and study groups was 22.5 ± 0.4 and 27.0 ± 0.4 months.Conclusions. As a result of a comparative analysis of the detection of adverse events and complications of the treatment, patients of the study group underwent treatment much easier than patients of the control group – in patients in the group of systemic chemotherapy, moderate and severe complications were detected in 44.4 % of cases, in the study group – in 2.5 % of cases. According to the results of a clinical study, regional chemotherapy is an effective method of treating patients with colon cancer metastases in the liver that are not controlled by systemic chemotherapy and is associated with a statistically significant increase in overall survival (p &lt; 0.05). For a more detailed study of the benefits of regional chemotherapy in this category of patients, further prospective clinical studies are necessary.

Ro 90-7501 suppresses colon cancer progression by inducing immunogenic cell death and promoting RIG-1-mediated autophagy

Colon cancer is one of the leading causes of cancer-related mortality worldwide. Current treatments, including surgery, chemotherapy, and radiation therapy, often have limited efficacy due to tumor heterogeneity and resistance mechanisms. Therefore, there is a critical need for novel therapeutic strategies that can enhance the immune response against colon cancer cells while promoting their efficient clearance. In this study, we investigated the anti-tumor effects of Ro 90-7501, a specific small molecule, in colon cancer cell lines (HCT8 and SW480) and in vivo models. MTS, EdU, Scratch Wound and Transwell assays were performed to detect the cell proliferation and metastasis. Additionally, flow cytometry and immunofluorescence staining were used to analyze changes in apoptosis and necrosis. Furthermore, we examined its ability to induce immunogenic cell death (ICD) and promote retinoic acid-inducible gene I (RIG-I)-mediated autophagy. The expression levels of key molecules involved in ICD and autophagy were assessed using Western blot analysis, immunofluorescence staining, and enzyme-linked immunosorbent assay (ELISA). Our findings demonstrated that Ro 90-7501 significantly suppressed colon cancer cell proliferation and metastasis, inducing apoptosis and necrosis. Mechanistically, Ro 90-7501 triggered ICD by upregulating the exposure of calreticulin (CRT) on the cell surface and increasing the release of high mobility group box 1 (HMGB1) and extracellular ATP levels. Concurrently, it promoted RIG-I-mediated autophagy via the MAVS-TRAF6 signaling axis, evidenced by increased expression of autophagy-related genes, such as microtubule-associated protein 1 light chain 3 (LC3) and Beclin 1 proteins, coupled with a reduction in P62 protein. Additionally, Ro 90-7501 treatment activated the RIG-I-MAVS-TRAF6 signaling axis in colon cancer cells. Furthermore, Ro 90-7501 enhanced the secretion of pro-inflammatory cytokines, such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNFα), thereby activating the immune response. In conclusion, Ro 90-7501 exhibits potent anti-tumor activity against colon cancer by inducing ICD and promoting RIG-I-mediated autophagy. These results suggest that Ro 90-7501 may serve as a promising therapeutic candidate for colon cancer treatment by enhancing both intrinsic cellular processes and adaptive immune responses. Further studies are warranted to elucidate the detailed mechanisms underlying these effects and to evaluate its therapeutic potential in clinical settings.

Metastasis of colon cancer requires Dickkopf-2 to generate cancer cells with Paneth cell properties.

Metastasis is the leading cause of cancer-related mortality. Paneth cells provide stem cell niche factors in homeostatic conditions, but the underlying mechanisms of cancer stem cell niche development are unclear. Here, we report that Dickkopf-2 (DKK2) is essential for the generation of cancer cells with Paneth cell properties during colon cancer metastasis. Splenic injection of Dkk2 knockout (KO) cancer organoids into C57BL/6 mice resulted in a significant reduction of liver metastases. Transcriptome analysis showed reduction of Paneth cell markers such as lysozymes in KO organoids. Single-cell RNA sequencing analyses of murine metastasized colon cancer cells and patient samples identified the presence of lysozyme positive cells with Paneth cell properties including enhanced glycolysis. Further analyses of transcriptome and chromatin accessibility suggested hepatocyte nuclear factor 4 alpha (HNF4A) as a downstream target of DKK2. Chromatin immunoprecipitation followed by sequencing analysis revealed that HNF4A binds to the promoter region of Sox9, a well-known transcription factor for Paneth cell differentiation. In the liver metastatic foci, DKK2 knockout rescued HNF4A protein levels followed by reduction of lysozyme positive cancer cells. Taken together, DKK2-mediated reduction of HNF4A protein promotes the generation of lysozyme positive cancer cells with Paneth cell properties in the metastasized colon cancers.

Metastasis of colon cancer requires Dickkopf-2 to generate cancer cells with Paneth cell properties

Metastasis is the leading cause of cancer-related mortality. Paneth cells provide stem cell niche factors in homeostatic conditions, but the underlying mechanisms of cancer stem cell niche development are unclear. Here, we report that Dickkopf-2 (DKK2) is essential for the generation of cancer cells with Paneth cell properties during colon cancer metastasis. Splenic injection of Dkk2 knockout (KO) cancer organoids into C57BL/6 mice resulted in a significant reduction of liver metastases. Transcriptome analysis showed reduction of Paneth cell markers such as lysozymes in KO organoids. Single-cell RNA sequencing analyses of murine metastasized colon cancer cells and patient samples identified the presence of lysozyme positive cells with Paneth cell properties including enhanced glycolysis. Further analyses of transcriptome and chromatin accessibility suggested hepatocyte nuclear factor 4 alpha (HNF4A) as a downstream target of DKK2. Chromatin immunoprecipitation followed by sequencing analysis revealed that HNF4A binds to the promoter region of Sox9, a well-known transcription factor for Paneth cell differentiation. In the liver metastatic foci, DKK2 knockout rescued HNF4A protein levels followed by reduction of lysozyme positive cancer cells. Taken together, DKK2-mediated reduction of HNF4A protein promotes the generation of lysozyme positive cancer cells with Paneth cell properties in the metastasized colon cancers.