High Cyclooxygenase-2 Expression Research Articles

In Vitro Screening of Lactic Acid Bacteria with RAW264.7 Macrophages and the Immunoregulatory Mechanism

Lactic acid bacteria (LAB) are commonly consumed as probiotics to improve gut barrier function and boost the immune system. This study aimed to screen LAB with high immunomodulatory activity using RAW264.7 macrophages. According to the results, Limosilactobacillus reuteri AUc2301 was selected from 84 screened strains that can stimulate RAW264.7 cell proliferation. Limosilactobacillus reuteri AUc2301 significantly enhanced the phagocytosis activity of RAW264.7 cells. In the ELISA test, Limosilactobacillus reuteri AUc2301 significantly promoted the release of interleukin-6, IL-1β, the tumor necrosis factor, and nitric oxide in RAW264.7 macrophages. In addition, Limosilactobacillus reuteri AUc2301 significantly inhibited the excessive release of IL-6, IL-1β, TNF-α, prostaglandin E2 as well as NO and the high expression of cyclooxygenase-2 in RAW264.7 macrophages induced by lipopolysaccharide. In further mechanism studies, Limosilactobacillus reuteri AUc2301 could regulate the nuclear factor-κB signaling pathway in RAW264.7 macrophages. Collectively, the screened Limosilactobacillus reuteri AUc2301 showed good immunomodulatory activity in vitro, and it has the potential to be developed as a novel probiotic.

Synergistic Mechanism of stir-baked Curcumae Radix with Vinegar in Dysmenorrhea Rats Based on UPLC-Q-TOF/MS Metabolomics

Curcumae Radix (i.e. Huangsiyujin: HSYJ), a well-known traditional Chinese medicine (TCM), has been widely used in clinical practice for many years to treat depression and primary dysmenorrhea. Modern pharmacological researches have demonstrated its anti-inflammatory, antidepressant, and dysmenorrhea relief effects. According to the processing theory of TCM, it is believed that stir-baked HSYJ with vinegar may enhance the ability to disperse stagnant hepatoqi and alleviate pain. However, whether the vinegar concoction of HSYJ with vinegar can enhance the therapeutic effect on the Qi stagnation due to liver depression (LDQS) type of dysmenorrhea and what its mechanism is have not been well explained. Based on the processing drugs theory of "stir-baked with vinegar into liver", a metabolomic approach was used to investigate the therapeutic effect and mechanism of stir-baked HSYJ with vinegar to enhance the treatment of dysmenorrhea in rats. By establishing a rat model of dysmenorrhea of the “LDQS” type, observation of blood rheology, uterine pathological sections, COX-2 and OTR protein expression and other indicators; analysis of urinary metabolic changes in rats by UPLC-Q-TOF-MS technique, to compare the potential biomarkers and metabolic pathways in the treatment of dysmenorrhea due to "liver stagnation and qi stagnation" before and after stir-baked HSYJ with vinegar. Stir-baked HSYJ with vinegar significantly inhibited the writhing response of rats, improved blood rheology, repaired damaged diseased uterus and inhibited high expression of COX-2 and OTR proteins in uterus; 68 differential metabolites were screened from the urine of rats, compared with the raw HSYJ, the levels of 14 metabolites were significantly changed in stir-baked HSYJ with vinegar, involving the pathways of phenylalanine, tyrosine and tryptophan metabolism, cysteine and methionine metabolism, aspartate and glutamate metabolism. The potentiating effect of stir-baked HSYJ with vinegar may be related to the regulation of multiple amino acid metabolic pathways.

Loss of CDX2 and high COX2 (PTGS2) expression in metastatic colorectal cancer.

Lack of expression of the tumour suppressor gene caudal-type homeobox 2 (CDX2) associates with poor outcomes in early stage colorectal cancer (CRC). Yet its prognostic value in the context of other prognostic biomarkers in metastatic CRC (mCRC) is unknown. Overexpressed cyclooxygenase-2 (COX2) has been reported in advanced CRC. However, CDX2 and COX2 relationship in mCRC remains undetermined. We aimed to assess their expression in mCRC tumours from a clinically characterised cohort and their influence on overall survival (OS) and progression-free survival (PFS) in first line. Among 720 consecutive mCRC patients, 346 had tumour samples appropriate for tissue microarray assembly and immunohistochemistry analyses. Clinical and survival data were retrospectively assessed. Loss of CDX2 expression was detected in 27 (7.8%) samples, enriched in poorly differentiated tumours (20%; p < 0.01) and in those with the BRAF p.V600E variant (40%; p < 0.01). Most tumours (93.4%) expressed COX2. COX2-negative samples were enriched in poorly differentiated mCRC. In unadjusted analyses, median OS (p < 0.001) and median PFS (p < 0.05) were inferior for patients with CDX2-negative versus CDX2-positive tumours. In conclusion, loss of CDX2 was significantly associated with poorly differentiated mCRC and BRAF p.V600E allele and a prognostic marker of worse OS.

Gene expression of cardiovascular risk markers in mononuclear cells of pregnant woman in relation to plasma leptin and homocysteine levels: A cross sectional study.

To investigate the relationship between anthropometric, biochemical, and hematologic parameters and serum leptin and homocysteine (Hcy) levels. Also, to determine the effect of leptin and Hcy on expression of genes associated with cardiovascular disease susceptibility (APOA1, LRP1, COX-1, and COX-2) in mononuclear cells of healthy pregnant women. Between August 2018 and January 2020, a cross-sectional study was conducted on 161 healthy pregnant women in Tabasco, southeastern Mexico. The study population was classified by trimester, according to gestational pregnancy. Anthropometric, biochemical (leptin and homocysteine), and hematologic data were obtained under fasting conditions. APOA1, LRP1, COX-1, and COX-2 expression in mononuclear cells was evaluated using RT-qPCR. Red cell indices (hemoglobin, hematocrit, and erythrocytes) were negatively and positively correlated with leptin and Hcy levels, respectively, in the first- and second-trimester groups. Increased leptin levels and low red cell indices were significantly associated with BMI <25.0 in the second-trimester group; however, no significant differences were observed in Hcy levels. Increased leptin and Hcy levels were significantly associated with high lipid indicators in the first- and third-trimester groups, respectively. High APOA1 and COX-2 expression was significantly associated with reduced leptin and increased Hcy levels in the second- and third-trimester groups. Increased leptin and Hcy levels during pregnancy, mainly associated with modifications in erythrocytes and lipid indices, may lead to early modification of genes related to lipid metabolism (APOA1) and proinflammatory response (COX-2) and, thereby, increase cardiovascular disease risk.

Advancements in the study of allergen-specific Th2 cell surface markers

Allergen-specific Th2 cells refer to a subset of Th2 cells that undergo substantial expansion following allergen stimulation. They play a crucial role in allergic diseases, and an increasing amount of research has revealed a close relationship between surface molecules on allergen-specific Th2 cells and allergic diseases. In comparison to other CD4+T cells or Th2 cells, allergen-specific Th2 cells exhibit low expression of CD27 but high expression of CD154, CD69, CRTH2, CD161, ST2, hPGDs, CD49d, and COX-2. They can be used for the identification of allergen-specific Th2 cells and serve as potential targets for the prevention and treatment of specific diseases. They hold significant value in preventing the onset and exacerbation of allergic diseases.

Inactivation of TRPM7 Kinase Targets AKT Signaling and Cyclooxygenase-2 Expression in Human CML Cells.

Cyclooxygenase-2 (COX-2) is a key regulator of inflammation. High constitutive COX-2 expression enhances survival and proliferation of cancer cells, and adversely impacts antitumor immunity. The expression of COX-2 is modulated by various signaling pathways. Recently, we identified the melastatin-like transient-receptor-potential-7 (TRPM7) channel-kinase as modulator of immune homeostasis. TRPM7 protein is essential for leukocyte proliferation and differentiation, and upregulated in several cancers. It comprises of a cation channel and an atypical α-kinase, linked to inflammatory cell signals and associated with hallmarks of tumor progression. A role in leukemia has not been established, and signaling pathways are yet to be deciphered. We show that inhibiting TRPM7 channel-kinase in chronic myeloid leukemia (CML) cells results in reduced constitutive COX-2 expression. By utilizing a CML-derived cell line, HAP1, harboring CRISPR/Cas9-mediated TRPM7 knockout, or a point mutation inactivating TRPM7 kinase, we could link this to reduced activation of AKT serine/threonine kinase and mothers against decapentaplegic homolog 2 (SMAD2). We identified AKT as a direct in vitro substrate of TRPM7 kinase. Pharmacologic blockade of TRPM7 in wildtype HAP1 cells confirmed the effect on COX-2 via altered AKT signaling. Addition of an AKT activator on TRPM7 kinase-dead cells reconstituted the wildtype phenotype. Inhibition of TRPM7 resulted in reduced phosphorylation of AKT and diminished COX-2 expression in peripheral blood mononuclear cells derived from CML patients, and reduced proliferation in patient-derived CD34+ cells. These results highlight a role of TRPM7 kinase in AKT-driven COX-2 expression and suggest a beneficial potential of TRPM7 blockade in COX-2-related inflammation and malignancy.

COX2-Inhibitory and Cytotoxic Activities of Phytoconstituents of Matricaria chamomilla L.

Chamomile tea is a popular beverage and herbal remedy with various health benefits, including antioxidant and antimicrobial activities and beneficial effects on metabolism. In this study, we investigated the inhibitory activities of secondary metabolites from Matricaria chamomile L. against COX2, an enzyme involved in inflammation and linked to cancer development. The cytotoxicity of the compounds was also evaluated on a panel of 60 cancer cell lines. Myricetin, one of the COX2-inhibiting and cytotoxic compounds in chamomile tea, was further studied to determine a proteomic expression profile that predicts the sensitivity or resistance of tumor cell lines to this compound. The expression of classical mechanisms of anticancer drug resistance did not affect the responsiveness of cancer cells to myricetin, e.g., ATP-binding cassette (ABC) transporters (ABCB, ABCB5, ABCC1, ABCG2), tumor suppressors (p53, WT1), and oncogenes (EGFR, RAS), whereas significant correlations between myricetin responsiveness and GSTP expression and cellular proliferation rates were observed. Additionally, Kaplan–Meier survival time analyses revealed that high COX2 expression is associated with a worse survival prognosis in renal clear cell carcinoma patients, suggesting a potential utility for COX2 inhibition by myricetin in this tumor type. Overall, this study provides insight into the molecular modes of action of chamomile secondary metabolites and their potential as cancer-preventive or therapeutic agents.

Celecoxib abrogates concanavalin A-induced hepatitis in mice: Possible involvement of Nrf2/HO-1, JNK signaling pathways and COX-2 expression.

Concanavalin A (ConA) is an established model for inducing autoimmune hepatitis (AIH) in mice, mimicking clinical features in human. The aimof the current study is to explore the possible protective effect of celecoxib, a cyclooxygenase-2 inhibitor,on immunological responses elicited in the ConA model of acute hepatitis. ConA (20mg/kg) was administered intravenously to adult male mice for 6h. Prior to ConA intoxication, mice in the treatedgroups received daily doses of celecoxib (30 and 60mg/kg in CMC) for 7days. Results revealed that administration of celecoxib 60mg/kg for 7days significantly protected the liver from ConA-induced liver damage revealed by significant decrease in ALT and AST serum levels. Celecoxib 30 and 60mg/kg pretreatment enhanced oxidant/antioxidant hemostasis by significantreduction of MDA and NO content and increase hepatic GSH contents and SOD activity. In addition, celecoxib 30 and 60mg/kg caused significant increase in hepatic nuclear factor erythroid 2-related factor 2 (Nrf2) and the stress protein heme oxygenase-1 (HO-1) levels. Moreover, celecoxib 30 and 60mg/kg inhibited the release of proinflammatory markers including IL-1β and TNF-α along with significant decrease in p-JNK, AKT phosphorylation ratio and caspase-3 expression. Besides, Con A was correlated to high expression of cyclooxygenase COX-2 and this increasing was improved by administration of celecoxib. These changes were in good agreement with improvement in histological deterioration. The protective effect of celecoxib was also associated with significant reduction of autophagy biomarkers (Beclin-1 and LC3II). In conclusion, celecoxib showed antioxidant, anti-inflammatory, anti-apoptotic and anti-autophagy activity against Con A-induced immune-mediated hepatitis. These effects could be produced by modulation of Nrf2/HO-1, IL-1B /p-JNK/p-AKT, JNK/caspase-3, and Beclin-1/LC3II signaling pathways.

Rutin Inhibits Hepatic and Pancreatic Cancer Cell Proliferation by Inhibiting CYP3A4 and GST

Abstract: Background: Digestive cancer is among the major causes of mortality and morbidity worldwide. Rutin, a bioactive secondary metabolite belonging to flavonoids and distributed in many fruits and vegetables has shown anti-proliferative, anti-cancer, and neuroprotective activities. In this study, the antiproliferative, antioxidant and apoptotic activities of rutin on hepatic and pancreatic cancer cell lines were investigated. Materials and Methods: The effect on cellular viability was monitored by SRB assay. Increasing activity of caspases (3/7) was used as an indicator of apoptosis. Additionally, the anti-inflammatory and antioxidant activities of rutin were evaluated after measuring amount of prostaglandin E2 (PGE2) produced and through DPPH free radical scavenging assays, respectively. Moreover, the inhibitory effect on both CYP3A4 and GST enzymes has also been evaluated. Results: According to the data presented here, rutin has anti-proliferative effect and raises the number of caspases 3/7 in investigated cell lines. Conclusion: HepG-2 cells showed the highest cellular growth inhibition, followed by BxPC-3, Huh-7, MiaPaCa-2, Suit-2, and the normal cell line HPDE. Rutin also inhibited the cyctochrome P450 enzyme (CYP450 3A4) and glutatihione-S-transferase activity, with dose-dependent inhibition. Furthermore, suppression of PGE2 synthesis in BxPC-3 cells supported rutin's anti-inflammatory action (high COX-2 expression). Keywords: Rutin, Pancreatic and Liver cancer, Apoptosis and Anti-inflammatory, Antioxidant.

A phase 1 study of TPST-1495 as a single agent and in combination with pembrolizumab in patients with advanced solid tumors.

3107 Background: TPST-1495 is a novel oral therapy designed to be a selective dual antagonist of the prostanoid receptors, EP2 and EP4. Through these receptors, PGE2 stimulates tumor growth, suppresses immunity, and supports adaptive resistance to checkpoint inhibitors. In preclinical models, dual EP2/EP4 inhibition with TPST-1495 increased anti-tumor efficacy, reduced immune suppression, and activated immune effector response significantly better than single EP2 or EP4 antagonists and the COX-2 inhibitor celecoxib. Methods: In this Phase 1 study, subjects with advanced solid tumors received TPST-1495 in a modified 3+3 design optimizing dose and schedule as monotherapy and in combination with standard dose pembrolizumab. Study objectives included safety, PK/PD, recommended phase 2 dose (RP2D), anti-tumor activity and exploratory biomarkers. AEs were assessed per CTCAE v5 and efficacy per RECIST v1.1. Results: TPST-1495 monotherapy was dosed BID (n=11), QD (n=22) or QD days 1-5 every 7 days (n=17) and in combination was dosed QD only with pembrolizumab 200 mg IV q3 weeks (n=24). TPST-1495 doses ranged from 15 mg QD to 100 mg BID. Most subjects had CRC (61%) followed by endometrial (5.4%) and pancreatic (5.4%) cancer. Median prior lines of systemic therapy were 4 (2-10) in monotherapy; 3 (1-8) in combination The most common treatment related AEs (TRAEs) were diarrhea (22%), abdominal pain (20%), fatigue, and nausea (18% each) for monotherapy and nausea (29%), fatigue (21%) and diarrhea (17%) for combination therapy. The most common Gr 3 TRAEs were anemia (6.0%) in monotherapy, and ALT/ AST increased and fatigue (4.2% each) in combination. No Gr 4/5 TRAEs were reported. For mono vs combo cohorts, the median time on study was 1.9 mo (0.2 – 10.2) vs 2.7 mo (0.5 – 10.4) and the disease control rate was 43% (19/44) vs 43% (9/21) with 1 PR in a combination subject with MSS CRC. Pharmacodynamic activities included on-treatment increases in urine PGE2 metabolites and drug-dependent TNF-α rescue in an ex-vivo whole blood PGE2 challenge assay. The RP2D is 50 mg QD for mono and in evaluation for combo therapy. In a subject with endometrial cancer with 22% tumor shrinkage and 222+ days on combination therapy paired biopsies showed high COX-2 expression in the TME at baseline and increased CD8+ and CD8+GrB T cell infiltration on treatment. Conclusions: PGE2 is an important target for cancer and immunotherapy but remains ineffectively drugged. TPST-1495 is a novel oral therapy that inhibits signaling only at the tumor promoting EP2/EP4 receptors. In patients with heavily treated solid tumors, primarily MSS CRC, TPST-1495 had pharmacodynamic activity, a manageable safety profile, and a potential signal of activity consistent with the preclinical data and IO combination mechanism. A combination therapy cohort in endometrial cancer is underway. Updated results and biomarker work will be presented. Clinical trial information: NCT04344795 .

TU-100 Antagonizes the M2 Polarization Phenotype of Macrophages in the Tumor Microenvironment by Suppressing the TLR4/NF-B/STAT3 Axis.

Macrophages are the most abundant immune cells in the tumor stroma, and their polarization states within the tumor microenvironment (TME) exert critical roles in tumorigenesis. TU-100 (Daikenchuto) is a commonly prescribed Japanese herbal medicine that has shown anti-cancer effects by regulating cancer-associated fibroblasts (CAFs) in the TME. However, its effects on tumor-associated macrophages (TAMs) remain unclear. TAMs were generated by macrophage exposure to tumor-conditioned medium (CM), and their polarization states were evaluated after TU-100 treatment. The underlying mechanism was further studied. TU-100 exhibited little cytotoxicity over a range of doses in M0 macrophages and TAMs. However, it could antagonize the M2-like polarization of macrophages evoked by tumor-CM exposure. These effects might be caused by the inhibition of TLR4/NF-B/STAT3 signaling in the M2-like phenotype of macrophages. Interestingly, TU-100 antagonized the malignancy promoting effects of M2 macrophages on hepatocellular carcinoma cell lines in vitro. Mechanistically, the administration of TU-100 restrained the high expression of MMP-2, COX-2, and VEGF in TAMs. TU-100 may alleviate the progression of cancer by regulating the M2 polarization of macrophages within the TME, suggesting a viable therapeutic approach.

Immunohistochemical assessment of COX2, P53, and beta-catenin expression to predict metachronous lesion development following polypectomy.

234 Background: Following colorectal polypectomy, 20-50% develop metachronous polyps, and some have increased colorectal cancer (CRC) risk. Surveillance colonoscopy is recommended to high-risk patients, based on index polyp number, histology and size. We aimed to assess if COX2, p53 and beta-catenin expression could predict metachronous polyp/ CRC risk. Methods: Tissue microarrays (TMA’s) were constructed for 339 patients from the INCISE study, a retrospective analysis of all patients undergoing screening polypectomy (Apr’09-Dec’16) followed by surveillance (6months-6years) in Glasgow, UK. Epithelial COX2, p53 and beta-catenin expression were assessed for each patient’s most advanced index polyp using immunohistochemistry, quantified by QuPath software. Kaplan-Meier analysis, log-rank tests and univariate/ multivariate cox regression assessed the association between marker expression and time to metachronous polyp/ CRC detection. Results: Low nuclear p53 expression was associated with shorter time to metachronous polyp/ CRC detection (p=0.024). Low nuclear (p=0.012) and membranous (p=0.046) beta-catenin expression was associated with shorter time to metachronous polyp/ CRC detection. There was no association between cytoplasmic COX2 expression and time to metachronous polyp/ CRC detection overall (p=0.309), but high cytoplasmic COX2 expression was associated with shorter time to detection of advanced metachronous lesions (advanced colorectal polyps/ CRC) (p=0.011). A combined score using the two most powerful biomarkers (nuclear p53 and nuclear beta-catenin – both high/ one low/ both low) was also associated with shorter time to metachronous polyp/ CRC detection (p=0.011). Univariate and multivariate cox regression are shown. Conclusions: Index polyp number and combined nuclear p53/beta-catenin expression independently predicted time to metachronous polyp/ CRC detection. [Table: see text]

Utility of COX-2 as Immunohistochemical Prognostic Marker in Relation to Various Histopathological Parameters and TNM Staging in Colorectal Carcinoma

Introduction: The most prevalent neoplasm of the gastrointestinal system is colorectal carcinoma. After lung and breast carcinoma, malignant colorectal cancer appears to be characterised by inflammation. Among the numerous recognised indicators of inflammation, Cyclooxygenase-2 (COX-2) has been identified as playing a key role in the early phases of carcinogenesis. The link between higher expression of COX-2 and the early stages of carcinogenesis and cancer development implies that COX-2 might be a target for precancerous colorectal lesion imaging. Aim: To assess the expression of COX-2 in colorectal carcinoma and the association with various histopathological parameters and Tumour Node Metastasis (TNM) staging. Materials and Methods: The current study is an observational, cross-sectional, retrospective, and prospective study that will take place in the Histopathology and Immunohistochemistry unit of the Department of Pathology, Jawaharlal Nehru Medical College, Sawangi (Meghe), Wardha, Maharashtra, India. Sample collection will be done during the the period of august 2021 to july 2024. The study included approximately 60-65 resected specimens from confirmed and planned Colectomy, Hemicolectomy, and Proctocolectomy specimens received in the Department of General Pathology, JNMC. Observations and results will be collected from the immunohistochemistry study by using COX-2 as a prognostic marker in colorectal carcinoma and correlating it with the TNM staging. Statistical analysis will be made by chi-square test and regression analysis.

Clinicopathological and prognostic significance of COX-2 in glioma patients: a meta-analysis.

In recent years, cyclooxygenase-2 (COX-2) has been identified as a cancer stem cell (CSC) marker in gliomas. Nevertheless, the clinical and prognostic significance of COX-2 in glioma patients remains controversial. To evaluate the correlation of COX-2 with the prognosis in glioma patients. Eligible studies on this subject were included, and pooled odd ratios (ORs) and hazard ratios (HRs) with 95% confidence intervals (95%CIs) were estimated. Publication bias was assessed through funnel plots, and heterogeneity and sensitivity were analyzed as well. In the present study, 11 articles with a total of 641 patients were included. The high expression of COX-2 in glioma patients was negatively associated with overall survival (OS) (n = 11; HR = 2.26; 95%CI = 1.79-2.86), and the subgroup analysis showed no differences in OS between Asian (n = 5; HR = 2.16; 95%CI = 1.57-2.97) and non-Asian (n = 6; HR = 2.39; 95%CI = 1.69-3.38) glioma patients. The Begg funnel plots test indicated that there was no evident risk of publication bias in the meta-analysis. The present study suggests that COX-2 could be recommended as a useful pathological and prognostic biomarker in the clinical practice.

Correlation and significance of COX-2, Ki67, VEGF and other immune indexes with the growth of malignant pulmonary nodules

ObjectiveThis study intends to explore the factors affecting the growth of pulmonary nodules in the natural process by immunohistochemical method.Methods40 cases of pulmonary nodules followed up for more than 3 years were divided into growth group (n = 20) and stable group (n = 20). The expressions of cyclooxygenase-2 (COX-2), Ki67, vascular endothelial growth factor (VEGF), CD44V6, epidermal growth factor receptor (EGFR), double microsome 2 (MDM2) and transforming growth factor (TGF)-β1 in pulmonary nodules were detected by immunohistochemical method so as to explore the relationship between it and the growth of pulmonary nodules.ResultsCompared with stable pulmonary nodules, the positive rates of COX-2, Ki67 and VEGF in the growth group were 85%, 80% and 55%, respectively. There was significant difference between the stable group and the growth group (P < 0.05). The correlation between other indexes and the growth of pulmonary nodules was not statistically significant (Pcd44v6 = 0.104;PEGFR = 0.337; PMDM2 = 0.49; PTGF-β1 = 0.141). In the subgroup of patients with non-invasive lung cancer, there was a correlation between VEGF and the growth of pulmonary nodules (P < 0.05).ConclusionThe high expression of COX-2, Ki67 and VEGF proteins may be significantly related to the growth of pulmonary nodules, and VEGF may be an important factor affecting the growth of malignant pulmonary nodules. This study intends to provide a research direction for further searching for the essential causes of the growth of pulmonary nodules.

Anti-Ferroptotic Effects of bone Marrow Mesenchymal Stem Cell-Derived Extracellular Vesicles Loaded with Ferrostatin-1 in Cerebral ischemia-reperfusion Injury Associate with the GPX4/COX-2 Axis.

Accumulating evidence of the critical role of Ferrostatin-1 (Fer-1, ferroptosis inhibitor) in cerebral ischemia has intrigued us to explore the molecular mechanistic actions of Fer-1 delivery by bone marrow mesenchymal stem cells-derived extracellular vesicles (MSCs-EVs) in cerebral ischemia-reperfusion (I/R) injury. In vivo middle cerebral artery occlusion (MCAO) in mice and in vitro oxygen-glucose deprivation/reperfusion (OGD/R) in hippocampal neurons were developed to simulate cerebral I/R injury. After Fer-1 was confirmed to be successfully delivered by MSCs-EVs to neurons, we found that MSCs-EVs loaded with Fer-1 (MSCs-EVs/Fer-1) reduced neuron apoptosis and enhanced viability, along with curtailed inflammation and ferroptosis. The regulation of Fer-1 on GPX4/COX2 axis was predicted by bioinformatics study and validated by functional experiments. The in vivo experiments further confirmed that MSCs-EVs/Fer-1 ameliorated cerebral I/R injury in mice. Furthermore, poor expression of GPX4 and high expression of COX-2 were witnessed in cerebral I/R injury models. MSCs-EVs/Fer-1 exerted its protective effects against cerebral I/R injury by upregulating GPX4 expression and inhibiting COX-2 expression. Taken together, our study indicates that MSCs-EVs/Fer-1 may be an attractive therapeutic target for the treatment of cerebral I/R injury due to its anti-ferroptotic properties.

Transcript levels of COX-2, TNF-α, IL-6 and IL-10 in chronic obstructive pulmonary disease: An association with smoking and severity

Chronic obstructive pulmonary disease (COPD) is characterized by inflammation. Tobacco smoking plays an important role in inflammation and subsequent disease progression. Therefore, inflammatory biomarkers (COX-2, TNF-α, IL-6 and IL-10 genes) expression levels were marked in patients with and without smoking habits and the severity of COPD. The severity of COPD, spirometry results and pack-years of smoking were assessed. The mRNA levels of COX-2, TNF-α, IL-6 and IL-10 in peripheral blood mononuclear cells were evaluated using reverse transcriptase-polymerase chain reaction. Relative fold expression and Pearson correlations were established among the group variables. Patients had a higher expression of COX-2 followed by TNF-α and IL-6 (p&lt;0.000). In contrast, IL-10 expression was decreased by half in patients compared to controls (p&lt;0.0001). COX-2 (10.37-fold), IL-6 (11.31-fold) and TNF-α (6.86-fold) were highly expressed in smokers (P &lt;0.0001). In contrast, IL-10 expression was dramatically lower in smokers than non-smokers (p&lt;0.0001). With increasing smoking pack-years, COX-2, TNF-α and IL-6 expression were significantly increased, but IL-10 gene expression was decreased (p&lt;0.05). COX-2, TNF-α and IL-6 gene expression progressively increased with severity grade, but IL-10 expression was dramatically reduced (p&lt;0.0001). COX-2 was positively correlated with TNF-α and IL-6 (r=0.39, p&lt;0.0001) whereas COX-2 was moderately correlated with IL-10 (r=-0.62 and 0.63, p&lt;0.0001). The higher expression of COX-2, TNF-α and IL-6 in patients with COPD suggests that these patients will likely benefit from COX-2, TNF-α and IL-6 blockers. Smoking cessation reduces the burden and severity of COPD.

Immunohistochemical analysis of MMP-9 and COX-2 expression in carotid atherosclerotic plaques among patients undergoing carotid endarterectomy: A prospective study

BackgroundMatrix metalloproteinase-9 protein (MMP-9) and cyclooxygenase-2 (COX-2) proteins may have a role in remodelling of atherosclerotic plaques. We analysed and compared the radiological, histological and immunohistochemical characteristics of carotid atherosclerotic plaques between symptomatic and asymptomatic patients who underwent carotid endarterectomy (CEA). MethodsThis prospective single-blinded study included 31 patients (70 [64-75] years, 58% males, 42% symptomatic) who underwent CEA and a total of 155 carotid plaque sections that were analysed. Preoperative assessment and multimodality diagnostic imaging with magnetic resonance imaging (MRI) or computed tomography angiography (CTA), histological and immunohistochemical analyses of carotid plaques including the expression of MMP-9 and COX-2 proteins were performed. ResultsSymptomatic and asymptomatic patients did not significantly differ in respect to preoperative characteristics. Unstable plaques were detected in 12/13 (92.3%, p = 0.020) symptomatic patients using MRI or CTA. There was no perioperative mortality and perioperative outcomes were comparable in both groups. A significantly higher expression of MMP-9 in macrophages was observed among symptomatic patients (p = 0.020). ROC curve analysis showed statistically significant associations of both the higher intensity of COX-2 staining in CD68 PG-M1 positive macrophages (area under the curve [AUC]=0.701, p = 0.014) and higher MVD (AUC=0.821, p < 0.001) within the plaque with cerebrovascular symptoms. The expression of COX-2 and the intensity of COX-2 staining in macrophages within the unstable carotid plaques detected by preoperative MRI or CTA were significantly higher (76.1% vs. 40.0%, p = 0.038; 76.2% vs. 30.0%, p = 0.01, respectively). ConclusionsAdvanced non-invasive multimodality diagnostic imaging including MRI or CTA is reliable in differentiating unstable from stable carotid plaques. High expression of MMP-9 and COX-2 in macrophages within the symptomatic plaque is associated with increased risk of cerebrovascular complications. Trial RegistrationThis study has been registered at the ISRCTN registry (ID ISRCTN46536832), isrctn.org Identifier: https://www.isrctn.com/ISRCTN46536832

353P Loss of CDX2 and high COX2 (PTGS2) expression in stage IV colorectal cancer

Lack of expression of the tumor suppressor CDX2 is associated with stage II/III colorectal cancer (CRC) poor outcomes. Its potential prognostic and predictive values in metastatic CRC (mCRC), and correlation with other potential biomarkers are of great interest. Overexpression of the PTGS2 gene encoding cyclooxygenase 2 (COX2) has shown a significantly direct association with advanced CRC stages. Reduced CDX2 in CRC has been suggested to enhance NF-κB-mediated inflammatory response, upregulating COX2 expression.

Abstract 725: Biomarkers of circulating vitamin D status and expression of tumor molecular markers Ki67, p53, and COX2 among colorectal cancer cases in the Southern Community Cohort Study

Abstract Introduction: Epidemiologic studies and controlled trials provide evidence that higher 25-hydroxyvitaminD is associated with improved survival in colorectal cancer (CRC), although there are limited data in African Americans (AAs), who are at greater risk for CRC mortality and vitamin D deficiency. Herein, we report associations between vitamin D serum biomarkers collected at cohort entry up to 15 years prior to CRC onset and CRC tumor subtypes defined by high expression of tumor markers (Ki67, p53, and COX2) that represent cellular processes (proliferation, apoptosis, and inflammation) affected by vitamin D. Importantly, higher expression of these markers reflects greater risk for metastasis and CRC-specific mortality. Methods: Data arise from the Southern Community Cohort Study, a cohort from the southeastern United States where the majority of participants are AA. Incident CRC cases with tumor sample were analyzed (n=104). Expression of cellular Ki67, p53, and COX2 (% positivity) were measured and categorized by literature-defined high vs. low cut-points (20%, 10%, and 20%, respectively). Logistic models adjusted for age, race, and sex were used to measure associations between vitamin D biomarkers: 25-hydroxyvitaminD, vitamin D binding protein (VDBP), and calculated free 25-hydroxyvitaminD (vitamin D not bound by VDBP) and the odds of a more aggressive tumor subtype, defined by higher cellular expression. Associations were assessed in the full sample and the subsample of AAs (n=70). Results: There were 52 tumors with high Ki67 expression, 37 with high p53, and 50 with high COX2. Fifteen tumors had high expression of all three markers. Cases were 55.8±0.4 (mean±SE) years at enrollment (range 40-79). The median 25-hydroxyvitaminD was lower in cases with high p53 expressing tumors compared to low expression (13.5 vs. 18.0 ng/mL, respectively, p = .03). Median values for other biomarkers were similar by tumor subtype (p &amp;gt .05). 25-hydroxyvitaminD was not associated with having a tumor with high Ki67 expression (odds ratio (OR) per 1 SD increase [95% CI] = 1.29 [0.85-1.95], p-trend = .23), high p53 expression (0.80 [0.52-1.23], p-trend = .30), or high COX2 expression (1.29 [0.83-2.00], p = .25). We observed null associations for VDBP, and free 25-hydroxyvitaminD with all tumor subtypes. Results were similar in AAs, although higher VDBP was associated with lower odds of high Ki67 tumors (0.45 [0.21-0.97], p-trend = .04 per 1 SD increase). Conclusion: In this sample of predominantly AA CRC cases, we observed no association between vitamin D biomarkers in stored serum and subsequent more aggressive CRC molecular subtypes. More work is needed to characterize the relationship between vitamin D and expression of CRC tumor markers, which may help clarify the protective role of maintaining adequate vitamin D levels in association with CRC. Citation Format: Thomas Lawler, Timothy Su, Qiuyin Cai, Mark Steinwandel, Wei Zheng, William J. Blot, Shaneda Warren Andersen. Biomarkers of circulating vitamin D status and expression of tumor molecular markers Ki67, p53, and COX2 among colorectal cancer cases in the Southern Community Cohort Study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 725.