Unveiling esophageal cancer treatment mechanisms: network pharmacology and molecular docking of Physcion.

This study investigates the effects of Physcion on esophageal cancer and its possible mechanisms of action. Potential Physcion targets were identified using databases. Transcriptomic data from 17 esophageal cancer and adjacent tissues were analyzed to find differentially expressed genes, intersecting with potential targets to select 16 key genes. Their expression and distribution were evaluated in patient sequencing data. Diagnostic potential was assessed through differential gene expression and ROC curves. Pathway enrichment analysis was performed using KEGG, and molecular docking simulations were conducted to assess Physcion's binding affinity to key genes. In vitro assays complemented these findings. A total of 161 drug targets were identified, narrowing down to 16 pivotal genes. Expression patterns were examined across cell populations, and enrichment analysis showed significant PI3K/AKT pathway involvement. Molecular docking indicated strong binding of Physcion to HSP90AA1 and MMP2. In vitro assays confirmed Physcion's dose- and time-dependent impact on esophageal cancer cells, with significant DAPI staining effects. Physcion shows promise as a therapeutic agent for esophageal cancer. The study supports its potential for clinical development and future research in esophageal cancer treatment.

ARHGAP4 promotes colon cancer metastasis through the TGF-ß signaling pathway and may be associated with T cell exhaustion.

BACKGROUND: Colon cancer is a prevalent invasive neoplasm in the gastrointestinal system with a high degree of malignancy. Despite extensive research, the underlying mechanisms of its recurrence and metastasis remain elusive.Rho GTPase activating protein 4 (ARHGAP4), a member of the small GTPases protein family, may be closely related to tumor metastasis, and its expression is increased in colon cancer. However, the role of ARHGAP4 in colon cancer metastasis is uncertain. This study investigates the impact of ARHGAP4 on the metastasis of colon cancer cells. Our objective is to determine the role of ARHGAP4 in regulating the invasive behavior of colon cancer cells. METHODS: We downloaded colon adenocarcinoma (COAD) data from the Cancer Genome Atlas (TCGA), and performed differential analysis and survival analysis. By using the CIBERSORT algorithm, we evaluated the proportion of infiltrating immune cells in colon cancer. We further analyzed whether ARHGAP4 is associated with T cell exhaustion. Finally, we investigated the impact of ARHGAP4 knockdown on the migration and invasion of colon cancer cells through in vitro cell experiments. Additionally, we utilized western blotting to assess the expression of protein related to the TGF-ß signaling pathway and epithelial-mesenchymal transition (EMT). RESULTS: We found that ARHGAP4 is upregulated in colon cancer. Subsequent survival analysis revealed that the high-expression group had significantly lower survival rates compared to the low-expression group. Immune infiltration analysis showed that ARHGAP4 was not only positively correlated with CD8+ T cells, but also positively correlated with T cell exhaustion markers programmed cell death 1 (PDCD-1), cytotoxic T-lymphocyte associated protein 4 (CTLA-4), and lymphocyte activating 3 (LAG-3). In vitro cell experiments, the knockdown of ARHGAP4 inhibited the migration and invasion of colon cancer cells. Among EMT-related proteins, when ARHGAP4 was knocked down, the expression of E-cadherin was increased, while the expression of N-cadherin and Vimentin was decreased. Meanwhile, the expression of TGF-ß1, p-Smad2, and p-Smad3, which are associated with the TGF-ß/Smad pathway, all decreased. CONCLUSION: ARHGAP4 promotes colon cancer metastasis through the TGF-ß/Smad signaling pathway and may be associated with T cell exhaustion. It plays an important role in the progression of colon cancer and may serve as a potential target for diagnosis and treatment of colon cancer.

Downregulation of RCN1 inhibits esophageal squamous cell carcinoma progression and M2 macrophage polarization.

Reticulocalbin 1 (RCN1) is a calcium-binding protein involved in the regulation of calcium homeostasis in the endoplasmic reticulum. The aim of this study was to explore the clinical value and biological role of RCN1 in esophageal squamous cell carcinoma (ESCC). In addition, we investigated the effect of RCN1 on the polarization of tumor-associated macrophages (TAMs). The GSE53625 dataset from the Gene Expression Omnibus database was used to analyze the expression of RCN1 mRNA and its relationship with clinical value and immune cell infiltration. Immunohistochemistry was used to validate the expression of RCN1 and its correlation with clinicopathological characteristics. Subsequently, transwell and cell scratch assays were conducted to evaluate the migration and invasion abilities of ESCC cells. The expression levels of epithelial-mesenchymal transition (EMT)-related proteins were evaluated by western blot, while apoptosis was detected by flow cytometry and western blot. Additionally, qRT‒PCR was utilized to evaluate the role of RCN1 in macrophage polarization. RCN1 was significantly upregulated in ESCC tissues and was closely associated with lymphatic metastasis and a poor prognosis, and was an independent prognostic factor for ESCC in patients. Knockdown of RCN1 significantly inhibited the migration, invasion, and EMT of ESCC cells, and promoted cell apoptosis. In addition, RCN1 downregulation inhibited M2 polarization. RCN1 is upregulated in ESCC patients and is negatively correlated with patient prognosis. Knocking down RCN1 inhibits ESCC progression and M2 polarization. RCN1 can serve as a potential diagnostic and prognostic indicator for ESCC, and targeting RCN1 is a very promising therapeutic strategy.

OLFM2 promotes epithelial-mesenchymal transition, migration, and invasion in colorectal cancer through the TGF-ß/Smad signaling pathway.

BACKGROUND: Colorectal cancer (CRC) is an aggressive tumor of the gastrointestinal tract, which is a major public health concern worldwide. Despite numerous studies, the precise mechanism of metastasis behind its progression remains elusive. As a member of the containing olfactomedin domains protein family, olfactomedin 2 (OLFM2) may play a role in tumor metastasis. It is highly expressed in colorectal cancer, and its role in the metastasis of CRC is still unclear. As such, this study seeks to explore the function of OLFM2 on CRC metastasis and its potential mechanisms. METHODS: Real-time fluorescence quantitative PCR and western blotting were used to study the expression of OLFM2 in human CRC and adjacent normal tissues. Knockdown and overexpression OLFM2 cell lines were constructed using siRNA and overexpression plasmids to explore the role of OLFM2 in the migration and invasion of CRC through transwell, and wound healing experiments. Finally, the expression of epithelial-mesenchymal transition (EMT) -related proteins and TGF-ß/Smad signaling pathway-related proteins was investigated using western blotting. RESULTS: In this study, we observed an elevation of OLFM2 expression levels in CRC tissues. To investigate the function of OLFM2, we overexpressed and knocked down OLFM2. We discovered that OLFM2 knockdown inhibited migration and invasion of colon cancer cells. Furthermore, E-cadherin expression increased while N-cadherin and Vimentin expression were opposite. It is no surprise that overexpressing OLFM2 had the opposite effects. We also identified that OLFM2 knockdown resulted in reduced TGF-ßR1 and downstream molecules p-Smad2 and p-Smad3, which are related to the TGF-ß / Smad pathway. In contrast, overexpressing OLFM2 significantly boosted their expression levels. CONCLUSION: The protein OLFM2 has been identified as a crucial determinant in the progression of CRC. Its mechanism of action involves the facilitation of EMT through the TGF-ß/Smad signaling pathway. Given its pivotal role in CRC, OLFM2 has emerged as a promising diagnostic and therapeutic target for the disease. These results indicate the potential of OLFM2 as a valuable biomarker for CRC diagnosis and treatment and highlight the need for further research exploring its clinical significance.

UiO-66 derived ZrO2@C catalysts for the double-bond isomerization reaction of 2-butene.

1-Butene, as one of the widely used chemical raw materials, can be produced by the double bond isomerization of 2-butene. However, the current yield of the isomerization reaction is only up to 20% or so. It is therefore an urgent issue to develop novel catalysts with higher performances. In this work, a high-activity ZrO2@C catalyst that is derived from UiO-66(Zr) is fabricated. The catalyst is prepared by calcining the precursor UiO-66(Zr) at high temperature in nitrogen, and characterized by XRD, TG, BET, SEM/TEM, XPS and NH3-TPD. The results demonstrate that the calcination temperature has significant influences on the catalyst structure and performance. Regarding the catalyst ZrO2@C-500, the selectivity and yield of 1-butene are 94.0% and 35.1%, respectively. The high performance is due to multiple aspects, including the inherited octahedral morphology from parent UiO-66(Zr), suitable medium-strong acidic active sites and high surface area. The present work will lead to a better understanding of the ZrO2@C catalyst and guide the rational design of high-activity catalysts for the double bond isomerization of 2-butene to 1-butene.

Morphometric features of gastric mucosa in atrophic gastritis: A different pattern between corpus and antrum.

Atrophic gastritis can cause mucosa thinning, while detailed metrological evidence is lacking. We aimed to compare the morphological features of full-thickness gastric mucosa in antrum and corpus and evaluate the diagnostic performance for atrophy. Gastric cancer patients were prospectively enrolled (N = 401). Full-thickness gastric mucosa was obtained. Foveolar length, glandular length and musculus mucosae thickness were measured. Pathological assessment was conducted using the visual analogue scale of the updated Sydney system. Areas under the receiver operating characteristic curves (AUCs) were calculated for different atrophy degrees. In corpus mucosa, foveolar length and musculus mucosae thickness were positively correlated with the atrophy degree (spearman's correlation coefficient [rs] = 0.231 and 0.224, respectively, P < .05); glandular length and total mucosal thickness were negatively correlated (rs = -0.399 and -0.114, respectively, P < .05). Total mucosal thickness did not correlate with antral atrophy degree (P = .107). The AUCs of total mucosal thickness for corpus and antral atrophy were 0.570 (P < .05) and 0.592 (P < .05), respectively. The AUCs for corpus atrophy, moderate and severe, and severe atrophy were 0.570 (P < .05), 0.571 (P = .003), and 0.584 (P = .006), respectively. The corresponding AUCs for antral atrophy were 0.592 (P = .010), 0.548 (P = .140), and 0.521 (P = .533), respectively. The tendency for mucosal thickness to thin with atrophy occurred in the corpus rather than in the antrum. The diagnostic performance of corpus and antral mucosal thickness was limited for atrophy.

Development and validation of a prognostic model for mitophagy-related genes in colon adenocarcinoma: A study based on TCGA and GEO databases.

BACKGROUND: Mitophagy is used by eukaryotic cells to eliminate damaged mitochondria. The deregulation of this process can lead to an accumulation of dysfunctional mitochondria and is implicated in carcinogenesis and tumorigenesis. Despite increasing evidence that mitophagy is involved in the development of colon cancer, the role of mitophagy-related genes (MRGs) in colon adenocarcinoma (COAD) prognosis and treatment remains largely unknown. METHODS: Differential analysis was used to identify differentially expressed mitophagy-related genes associated with COAD and conduct key module screening. Cox regression and least absolute shrinkage selection operator, and other analyses were used to characterize prognosis-related genes and verify the feasibility of the model. The model was tested using GEO data and a nomogram was constructed for future clinical application. The level of immune cell infiltration and immunotherapy were compared between the two groups, and sensitivity to treatment with many commonly used chemotherapeutic agents was assessed in individuals with different risk factors. Finally, qualitative reverse transcription polymerase chain reaction and western blotting were performed to assess the expression of prognosis-related MRGs. RESULTS: A total of 461 differentially expressed genes were mined in COAD. Four prognostic genes, PPARGC1A, SLC6A1, EPHB2, and PPP1R17, were identified to construct a mitophagy-related gene signature. The feasibility of prognostic models was assessed using Kaplan-Meier analysis, time-dependent receiver operating characteristics, risk scores, Cox regression analysis, and principal component analysis. At 1, 3, and 5 years, the area under the receiver operating characteristic curves were 0.628, 0.678, and 0.755, respectively, for TCGA cohort, and 0.609, 0.634, and 0.640, respectively, for the GEO cohort. Drug sensitivity analysis found that camptothecin, paclitaxel, bleomycin, and doxorubicin were significantly different between low- and high-risk patients. The qPCR and western blotting results of clinical samples further confirmed the public database results. CONCLUSIONS: This study successfully constructed a mitophagy-related gene signature with significant predictive value for COAD, informing new possibilities for the treatment of this disease.

Reduced Adenoma Miss Rate With 9-Minute vs 6-Minute Withdrawal Times for Screening Colonoscopy: A Multicenter Randomized Tandem Trial.

INTRODUCTION: Although the 9-minute mean withdrawal time (m-WT) is often reported to be associated with the optimal adenoma detection rate (ADR), no randomized trials of screening colonoscopy have confirmed the impact of a 9-minute m-WT on adenoma miss rate (AMR) and ADR. METHODS: A multicenter tandem trial was conducted in 11 centers. Seven hundred thirty-three asymptomatic participants were randomized to receive segmental tandem screening colonoscopy with a 9-minute withdrawal, followed by a 6-minute withdrawal (9-minute-first group, 9MF, n = 366) or vice versa (6-minute-first group, 6MF, n = 367). The primary outcome was the lesion-level AMR. RESULTS: The intention-to-treat analysis revealed that 9MF significantly reduced the lesion-level (14.5% vs 36.6%, P < 0.001) and participant-level AMR (10.9% vs 25.9%, P < 0.001), advanced adenoma miss rate (AAMR, 5.3% vs 46.9%, P = 0.002), multiple adenomas miss rate (20.7% vs 56.5%, P = 0.01), and high-risk adenomas miss rate (14.6% vs 39.5%, P = 0.01) of 6MF without compromising detection efficiency ( P = 0.79). In addition, a lower false-negative rate for adenomas ( P = 0.002) and high-risk adenomas ( P < 0.05), and a lower rate of shortening surveillance schedule ( P < 0.001) were also found in 9MF, accompanying with an improved ADR in the 9-minute vs 6-minute m-WT (42.3% vs 33.5%, P = 0.02). The independent inverse association between m-WT and AMR remained significant even after adjusting ADR, and meanwhile, 9-minute m-WT was identified as an independent protector for AMR and AAMR. DISCUSSION: In addition to increasing ADR, 9-minute m-WT also significantly reduces the AMR and AAMR of screening colonoscopy without compromising detection efficiency.

A novel prognostic model based on urea cycle-related gene signature for colorectal cancer.

Background: Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the world. This study aimed to develop a urea cycle (UC)-related gene signature that provides a theoretical foundation for the prognosis and treatment of patients with CRC. Methods: Differentially expressed UC-related genes in CRC were confirmed using differential analysis and Venn diagrams. Univariate Cox and least absolute shrinkage and selection operator regression analyses were performed to identify UC-related prognostic genes. A UC-related signature was created and confirmed using distinct datasets. Independent prognostic predictors were authenticated using Cox analysis. The Cell-type Identification by Estimating Relative Subsets of RNA Transcripts algorithm and Spearman method were applied to probe the linkage between UC-related prognostic genes and tumor immune-infiltrating cells. The Human Protein Atlas database was used to determine the protein expression levels of prognostic genes in CRC and normal tissues. Verification of the expression levels of UC-related prognostic genes in clinical tissue samples was performed using real-time quantitative polymerase chain reaction (qPCR). Results: A total of 49 DEUCRGs in CRC were mined. Eight prognostic genes (TIMP1, FABP4, MMP3, MMP1, CD177, CA2, S100P, and SPP1) were identified to construct a UC-related gene signature. The signature was then affirmed using an external validation set. The risk score was demonstrated to be a credible independent prognostic predictor using Cox regression analysis. Functional enrichment analysis revealed that focal adhesion, ECM-receptor interaction, IL-17 signaling pathway, and nitrogen metabolism were associated with the UC-related gene signature. Immune infiltration and correlation analyses revealed a significant correlation between UC-related prognostic genes and differential immune cells between the two risk subgroups. Finally, the qPCR results of clinical samples further confirmed the results of the public database. Conclusion: Taken together, this study authenticated UC-related prognostic genes and developed a gene signature for the prognosis of CRC, which will be of great significance in the identification of prognostic molecular biomarkers, clinical prognosis prediction, and development of treatment strategies for patients with CRC.

Establishment and evaluation of a nomogram predicting risks of missed diagnoses of colorectal polyps.

BACKGROUND: A missed diagnosis of colorectal polyps during colonoscopy may be associated with the occurrence of interval colorectal cancer. The risk factors for a missed diagnosis or a method to predict the risk of a missed diagnosis of colorectal polyps during colonoscopy remain unidentified. METHODS: The clinical data of patients who underwent two colonoscopies within three months at the Affiliated Hospital of North Sichuan Medical College between February 2017 and August 2019 were retrospectively reviewed. Independent risk factors for missed diagnoses were identified, and a nomogram was established to predict the risk of missed diagnoses. The prediction performance of the nomogram was evaluated using C-index and calibration curves, and its clinical application value was assessed using the Youden index and decision curve analysis. RESULTS: Independent influencing factors for missed diagnoses included age, endoscopist experience, bowel preparation, retroflected view, withdrawal time, number of polyps in the right colon, and number of polyps ≥ 6 mm. The C-index of the nomogram in the training and validation cohorts was 0.763 (95% confidence interval [CI]: 0.724 - 0.807) and 0.726 (95%CI: 0.657 - 0.794), respectively. The optimal cut-off value of the nomogram calculated using the Youden index was 152.2 points. Under the cut-off value, the sensitivity, specificity, positive predictive value, and negative predictive value were 67.1%, 75.7%, 45.8%, and 88.2%, respectively, in the training cohort, and 57.1%, 79.9%, 53.3%, and 82.3%, respectively, in the validation cohort. CONCLUSIONS: The nomogram provides a reference value for clinicians to analyse the risk of a missed diagnosis of colorectal polyps in individuals, identify high-risk groups, and formulate appropriate follow-up strategies.

Twenty-year span of global acute pancreatitis trends: A bibliometric analysis.

OBJECTIVE: This study aims to investigate the global research routine and trends of acute pancreatitis over the last twenty years based on the production, hotspots, and frontiers of published articles as well as to provide the global health system with a bibliometric reference. METHODS: The Web of Science core collection database was retrieved for acute pancreatitis original articles and review articles published from January 1, 1999 to May 17, 2020. Duplicates and discrete papers were excluded. Articles were evaluated for several characteristics including number of citations, publication time, country of origin, institution, journal and authorship. RESULTS: A total of 7001 articles originated from 94 countries and were published in 1263 journals. The China contributed most articles (1752) followed by USA (1214). The research was major published in specialized journals including the Pancreas (511) and pancreatology (351). Universities were the main institutions of science progress. High-impact articles focused on the fields of clinical medicine. A steady growth was observed in the last 20 years from 1999 to 2020. CONCLUSION: This comprehensive bibliometric study indicates that severe acute pancreatitis and necrotizing pancreatitis are significant topic in the acute pancreatitis research. The structured information may be helpful in understanding research trends, and locating research hot spots and gaps in this domain.

Long-term outcomes of endoscopic treatment for colorectal laterally spreading tumor: a large-scale multicenter retrospective study from China.

BACKGROUND: Laterally spreading tumor (LST) is a type of precancerous lesion of colorectal cancer with high malignant potential. The present study aimed to evaluate long-term outcomes of endoscopic treatment for LST in Chinese patients. METHODS: This study was a retrospective review of data collected from 653 included patients with LST from six regional representative hospitals in China between January 2007 and January 2017. Demographic characteristics, endoscopic features of LST, operation-related data, and follow-up results were collected and analyzed. RESULTS: LST-granular type (LST-G, 80.3%) was much more common than LST-non-grandular type (LST-NG, 19.7%). The overall submucosal invasion rate of all LSTs was 6.1% and the submucosal invasion rate of LST-NG was significantly higher than that of LST-G (6.79% vs. 3.87%, p = 0.000). The en bloc resection rate of ESD and EMR treatment was 96% and 93.7%, respectively, with pathologic R0 resection rate of 90.1% and 82.8%. After an average duration of follow-up about 34.52 ± 11.76 months, the recurrence rate of ESD was 3.47%, and the recurrence rate of EMR was 8.8% after an average follow-up of about 38.44 ± 4.42 months. However, the recurrence rate of ESD was much lower than piecemeal EMR for LST (3.47% vs. 8.62%, p = 0.017). Retroflexion-assisted technique applied for resection of rectal LST was associated with a significantly shortened operating time (85.40 min vs. 174.18 min, p = 0.002). CONCLUSION: Endoscopic resection is a safe and efficient modality for the treatment of colorectal LST with a relatively low recurrence rate and shortened operating time with the use of retroflexion.

NLRP3 inflammasome inhibitor CY-09 reduces hepatic steatosis in experimental NAFLD mice.

AIMS: Non-alcoholic fatty liver disease (NAFLD) has become one of the most common chronic liver diseases. The NOD-like receptor protein 3 (NLRP3) inflammasome was suggested to be involved in the pathogenesis of NAFLD. A small-molecule named CY-09 is a new selective and direct inhibitor of the NLRP3 inflammasome. We aimed to investigate whether CY-09 is effective for the treatment of NAFLD in a high-fat diet (HFD)-induced mouse model. METHODS: Twenty mice were fed by HFD for 14 weeks, and then were randomly assigned into two groups: (1) control group receiving dimethylsulfoxide (DMSO) solution; (2) CY-09 group receiving CY-09 injection. In an 8-week follow-up, oral glucose tolerance test (OGTT) and homeostasis model assessment of insulin resistance (HOMA-IR) were used to measure glucose metabolism. Liver steatosis was evaluated by the NAFLD activity score (NAS) and deemed as the primary outcome. RESULTS: The body weight in CY-09 group was significantly lower than the DMSO control group on 27 weeks (41.0 ± 3.5 g vs. 49.7 ± 5.2 g, P = 0.014). The area under the curve (AUC) of OGTT was less in CY-09 group than that in DMSO group (35.81 ± 6.79 vs. 22.91 ± 2.58 mmol/L·hr, P = 0.004), as well as HOMA-IR (14.36 ± 3.89 vs. 8.82 ± 2.04 mmol.mIU.L-2, P = 0.023). Microscopically, liver lipid droplets dramatically improved and significantly lower NAS was observed in CY-09 group (8.25 ± 1.26 vs. 3.20 ± 0.45, P < 0.001). CONCLUSION: CY-09 reduces hepatic steatosis in experimental NAFLD mice and CY-09 may be a potential therapeutic drug of NAFLD in clinical practice.

A huge bladder diverticulum in an elderly: A case report.

An 81-year-old male patient presented to the department of gastroenterology with increasing lower abdominal pain for 2 years, aggravated with bloody stool for 1 month. Computed tomographic examination revealed a huge cyst (207 × 93 × 208 mm3) in the abdominal cavity, absence of bladder, thickening and strengthening of the rectal wall, and benign prostatic hyperplasia. Colonoscopy showed colon cancer and surgery was planned. Interestingly, after magnetic resonance imaging and cystography, we found colon cancer and a large bladder diverticulum rather than tumor metastasis or others. Severe bacteremia occurred in the elderly chronic obstructive pulmonary disease patient before operation. After careful consideration, we decided to take a large risk and combined urology and gastrointestinal surgery professionals to carry out bladder diverticulectomy, cystostomy, radical resection of rectal carcinoma, and so on. Fortunately, the patient recovered well after the operation. In addition to the common tumor metastasis and cystadenoma, the abdominal mass should also be alert to the rare bladder diverticulum, which eventually leads to diagnostic confusion. Multidisciplinary diagnosis and treatment has become an important treatment for complex diseases.

Cantharidin suppresses gastric cancer cell migration/invasion by inhibiting the PI3K/Akt signaling pathway via CCAT1.

Cantharidin (CTD) is a traditional Chinese medicine that shows an anticancer effects in multiple types of cancer cells. However, the mechanism of CTD anti-cancer function in gastric cancer (GC) is still unclear. The aim of the present study was to investigate the underlying mechanism that CTD inhibits proliferation and migration through suppression of the PI3K/Akt signaling. CTD induced GC cell apoptosis and inhibited metastasis measured by CCK8 assays as well as wound healing assays and transwell assays. Mechanistic investigations suggested that CTD modulated the PI3K/Akt signaling via western-blot and quantitative q-PCR. In addition, we identified and confirmed CCAT1 as a novel direct target of CTD inhibited PI3K/AKt signaling expression. In conclusion, our results provide new point into the critical role of CTD in suppressing PI3K/Akt signaling via down-regulation of CCAT1, resulting in suppression GC cell growth and migration/invasion.

Autoinducer-2 of gut microbiota, a potential novel marker for human colorectal cancer, is associated with the activation of TNFSF9 signaling in macrophages.

Objectives: The interaction between the quorum sensing (QS) molecules of gut microbiota and the immunity of colorectal cancer (CRC) has not been investigated before. Methods: We measured the concentration of autoinducer-2 (AI-2) in samples of stool, colorectal tissue, saliva and serum of CRC patients, and compared this to AI-2 levels in colorectal adenoma (AD) and normal colon mucosa (NC). To explore the activated signaling pathways involved, we utilized AI-2 extracted from Fusobacterium nucleatum to stimulate macrophages and validated these in vitro findings in human CRC tissues. Results: The AI-2 concentration in both colorectal tissue and stool of CRC patients was significantly higher when compared to that in AD and NC (all P values < .01). The AI-2 concentration along with the progression of CRC in both tissues and stools was significantly increased (P= .045，P= .0003, respectively). After AI-2 stimulation, TNFSF9 was the most significantly increased protein in macrophage cells (P < .01). TNFSF9 expression was significantly higher in CRC tissues when compared to NCs (P< .0001), which was mainly derived from macrophages in the tumor microenvironment. Moreover, AI-2 level was positively associated with CD3 + T cell numbers (P= .0462), and negatively associated with CD4/CD8 ratio (P= .0113) within CRC tissues. Conclusions: We demonstrated for the first time that AI-2 may serve as a novel marker for screening CRC in the clinic. AI-2 was associated with tumor immunity in CRCs through tumor-associated macrophages and CD4/CD8 ratio in a TNFSF9-dependent manner.

Autoinducer-2 of Fusobacterium nucleatum promotes macrophage M1 polarization via TNFSF9/IL-1ß signaling.

The effect of Fusobacterium nucleatum (F. nucleatum) autoinducer-2 (AI-2) on the polarization of macrophages and the underlying mechanism is not known. We investigated the effect of F. nucleatum AI-2 on the migration and polarization of cultured macrophages. We further screened AI-2-interacting proteins in macrophages using a quantitative proteomics strategy, and evaluated the expression of TNFSF9/TRAF1/p-AKT/IL-1ß signaling in cultured macrophages and human colorectal cancer (CRC). The data showed that F. nucleatum AI-2 enhanced the mobility and M1 polarization of macrophages, possibly through TNFSF9/TRAF1/p-AKT/IL-1ß signaling. Moreover, TNFSF9 and IL-1ß expression was significantly increased in human CRCs when compared to normal colon (P < 0.05), and was associated with AI-2 concentration and increased survival. Together, our data suggested that AI-2 induced macrophage M1 polarization by activating the TNFSF9/IL-1ß pathway. Thus, AI-2 may serve as a promising novel target for immunotherapy of gut microbiota-related diseases.