Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: miRNA-503 inhibition exerts anticancer effects and reduces tumor growth in mesothelioma

doi: 10.1186/s13046-025-03283-0

Figure Lengend Snippet: Histological analysis of miR-503 target genes and transcriptomic analysis of miR-503 inhibition. A Evaluation of the expression of BTG1, CCNG1, EDG1 and TIMP2, in normal pleural (PL) and pleural mesothelioma tissues (EPM epithelial istotype; SPM, sacomatoid istotype). Yellow Scale bar = 100 μm. Data shown are mean ± S.D. of five independent experiments. B IHC analysis of the putative miR-503 target genes on mice tumors treated with LNPs-anti-miR-503 or empty LNPs (original magnification 20X). A and B confirm the in vitro results. C Transcriptome analysis in MSTO cells after miR-503 inhibition detected about 500 deregulated genes (red upregulated, green downregulated). D Expression analysis of MM biomarkers deregulated by inhibition of miR-503 in MM cells. E q-PCR analysis in human MM samples of the miR-503 MM biomarkers confirms the inverse expression only for CXCL8, SERPINE1 and SPP. F IHC analysis of the miR-503 deregulated MM biomarkers on mice tumors treated with LNPs-anti-miR-503 or empty LNPs (original magnification 20X). For each IHC panel bar plots report the score for the samples analysed. Data presented as the mean ± SD of at least three independent experiments ( n = 3) for the in vitro experiments while for the in vivo analysis numbers of samples are reported in Methods. * p < 0.05, ** p < 0.01 *** p < 0.001 **** p < 0.0001 vs. control

Article Snippet: The primary antibodies used were: Rabbit anti-human Ki67 (DAKO Agilent, Santa Clara, CA USA), Rabbit anti-human BTG1 (orb35408 Biorbyt, Durham, NC, USA), Rabbit anti-human CCNG1 (orb167206 Biorbyt), Rabbit anti-human S1PR1 (EDG1) (orb350684 Biorbyt), Rabbit anti-human TIMP2 (orb543218 Biorbyt), Rabbit anti-human CXCL8 (17038-1-AP Proteintech Rosemont, IL, USA), Rabbit anti-human Osteopontin, (SPP 20416-1-AP Proteintech), Rabbit anti-human SERPINE1 (PAI-1 #49536 Cell Signaling, Danvers, MA, USA).

Techniques: Inhibition, Expressing, In Vitro, In Vivo, Control

Journal: Discover Oncology

Article Title: Mechanism of CXCL8 regulation of methionine metabolism to promote angiogenesis in gliomas

doi: 10.1007/s12672-024-01467-2

Figure Lengend Snippet: Clinical information analysis of CXCL8 in gliomas. A Forest plot of the relationship between CXCL8 and prognosis in the TCGA glioma cohort. B CXCL8 expression plots of normal and tumor samples in the TCGA glioma cohort. C RT-PCR assay for detecting CXCL8 mRNA expression levels in glioma tissues and paracancer tissues. D Violin plots depicting the distribution of CXCL8 expression levels in TCGA glioma cohorts, stratified by different WHO staging categories for CXCL8 expression level distribution. E Kaplan–Meier curves of CXCL8 gene expression versus overall patient survival in glioma cohorts of TCGA and CGGA

Article Snippet: Reagents and antibodies: 1640 medium and fetal bovine serum were purchased from Gibco, trypsin was purchased from Zhejiang GinoSebail Biotechnology Co., Ltd, matrix gel was purchased from Shanghai Yisheng, CXCL8–CXCR2 signal axis inhibitor SB225002 (HY-16711) was purchased from MCE, bevacizumab was purchased from Roche, Germany, and antibodies against CXCL8 (94407), VEGFA (50661), and β-actin (4967) were purchased from Cell Signaling Technology, USA.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Gene Expression

Journal: Discover Oncology

Article Title: Mechanism of CXCL8 regulation of methionine metabolism to promote angiogenesis in gliomas

doi: 10.1007/s12672-024-01467-2

Figure Lengend Snippet: Effect of methionine-restricted environment on CXCL8 expression and angiogenesis. A Heat map of metabolomics analysis of targeted amino acids in control and methionine-restricted tolerant cell groups. B Statistical plots of the levels of l -methionine and its derivatives, N -acetyl- l -methionine, N -formyl- l -methionine, and methionine sulfoxide, for control and methionine-restricted tolerant cell groups (* P < 0.05,** P < 0.01,*** P < 0.001). C Western blotting to detect the expression of CXCL8 and VEGFA proteins in U251 cells. D RT-PCR to detect the expression level of CXCL8 mRNA in methionine-restricted cultured cells after 0, 4, 12, and 24 h (* P < 0.05). E Control and methionine-restricted diet group nude mice transplanted tumor tissues immunohistochemical staining of CD31, VEGFA, scale bar = 50 μm. F. Tubulogenesis assay to detect the effect of methionine-restricted culture on the tubulogenic ability of HUVEC, U87. G – L Graphs of statistical analysis of tube length, number of nodes, and segments length as compared with MSM-0d (* P < 0.05,** P < 0.01)

Article Snippet: Reagents and antibodies: 1640 medium and fetal bovine serum were purchased from Gibco, trypsin was purchased from Zhejiang GinoSebail Biotechnology Co., Ltd, matrix gel was purchased from Shanghai Yisheng, CXCL8–CXCR2 signal axis inhibitor SB225002 (HY-16711) was purchased from MCE, bevacizumab was purchased from Roche, Germany, and antibodies against CXCL8 (94407), VEGFA (50661), and β-actin (4967) were purchased from Cell Signaling Technology, USA.

Techniques: Expressing, Control, Western Blot, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Immunohistochemical staining, Staining

Journal: Discover Oncology

Article Title: Mechanism of CXCL8 regulation of methionine metabolism to promote angiogenesis in gliomas

doi: 10.1007/s12672-024-01467-2

Figure Lengend Snippet: Effect of inhibiting CXCL8 expression on glioma angiogenesis. A , B CAM experiments to determine the effect of CXCL8–CXCR2 signaling axis inhibitor SB225002 treatment on graft tumor angiogenesis. C Immunohistochemical staining of nude mice graft tumor tissues for CD31, VEGFA, scale bar = 50 μm. D YSM experiments to determine the effect of SB225002 and positive control bevacizumab treatment on angiogenesis

Article Snippet: Reagents and antibodies: 1640 medium and fetal bovine serum were purchased from Gibco, trypsin was purchased from Zhejiang GinoSebail Biotechnology Co., Ltd, matrix gel was purchased from Shanghai Yisheng, CXCL8–CXCR2 signal axis inhibitor SB225002 (HY-16711) was purchased from MCE, bevacizumab was purchased from Roche, Germany, and antibodies against CXCL8 (94407), VEGFA (50661), and β-actin (4967) were purchased from Cell Signaling Technology, USA.

Techniques: Expressing, Immunohistochemical staining, Staining, Positive Control

Journal: Discover Oncology

Article Title: Mechanism of CXCL8 regulation of methionine metabolism to promote angiogenesis in gliomas

doi: 10.1007/s12672-024-01467-2

Figure Lengend Snippet: Transcriptomic analysis of the inhibition of the CXCL8–CXCR2 signaling axis in methionine-restricted tolerant cells. A Heat map of differentially expressed genes in U251-M and U251-M cells treated with SB225002. B Volcano map of differentially expressed genes in U251-M cells and U251-M cells treated with SB225002. C GO enrichment analysis of differentially expressed genes in U251-M cells and GO enrichment analysis results of differentially expressed genes in U251-M cells treated with SB225002. D KEGG enrichment analysis results of differentially expressed genes in U251-M and U251-M cells treated with SB225002. E GSEA enrichment analysis showed that inhibiting of the CXCL8–CXCR2 signaling axis in U251-M glioma cells would activate ATP metabolic processes. F GSEA enrichment analysis showed that inhibition of the CXCL8–CXCR2 signaling axis in U251-M glioma cells activates negatively regulated cell migration and angiogenesis

Article Snippet: Reagents and antibodies: 1640 medium and fetal bovine serum were purchased from Gibco, trypsin was purchased from Zhejiang GinoSebail Biotechnology Co., Ltd, matrix gel was purchased from Shanghai Yisheng, CXCL8–CXCR2 signal axis inhibitor SB225002 (HY-16711) was purchased from MCE, bevacizumab was purchased from Roche, Germany, and antibodies against CXCL8 (94407), VEGFA (50661), and β-actin (4967) were purchased from Cell Signaling Technology, USA.

Techniques: Inhibition, Migration

Journal: Discover Oncology

Article Title: Mechanism of CXCL8 regulation of methionine metabolism to promote angiogenesis in gliomas

doi: 10.1007/s12672-024-01467-2

Figure Lengend Snippet: Targeted metabolomic analysis of the CXCL8-CXCR2 signaling axis inhibition in methionine-restricted tolerant cells. A – D Targeted amino acid metabolomic approach to detect the levels of l -methionine ( A ), methionine sulfoxide ( B ), N -acetyl- l -methionine ( C ), N -formyl- l -methionine ( D ) in U251-M cells and U251-M + SB225002 cells

Article Snippet: Reagents and antibodies: 1640 medium and fetal bovine serum were purchased from Gibco, trypsin was purchased from Zhejiang GinoSebail Biotechnology Co., Ltd, matrix gel was purchased from Shanghai Yisheng, CXCL8–CXCR2 signal axis inhibitor SB225002 (HY-16711) was purchased from MCE, bevacizumab was purchased from Roche, Germany, and antibodies against CXCL8 (94407), VEGFA (50661), and β-actin (4967) were purchased from Cell Signaling Technology, USA.

Techniques: Inhibition

Journal: iMeta

Article Title: Cell‐type‐specific expression analysis of liver transcriptomics with clinical parameters to decipher the cause of intrahepatic inflammation in chronic hepatitis B

doi: 10.1002/imt2.221

Figure Lengend Snippet: Chemokines and immune‐negative regulators in T cells and macrophages associated with liver injury. (A) Left: Heatmap of gene expression of immune regulation, chemokines, complements, cytokines, and interferons (rows) for all 82 patients (columns) in the four phases of CHB. Right: 95% credible intervals of ALT and AST slopes versus gene expression in Left. Colored bubbles indicate positive (red) or negative slopes (blue) (B) Left: Radar charts of the PSEA coefficient value ( β P ) for genes of different reference cell types in chronic hepatitis patients. Right: Changes in gene coefficients of different reference cell types comparing chronic hepatitis to HBeAg‐positive chronic infection. (C, D) Partial residual plot with fitted posterior lines and violin plot of slope distributions from the fitted lines in chronic hepatitis (red) and HBeAg‐positive chronic infection (blue) for gene expression contribution by T cells and iMac cells. Using the established PSEA model, we analyzed upregulated immune checkpoint genes and chemokine genes to discern their predominant expression cell types and expression variations across different clinical stages. (E) Our findings indicated that during the immune‐active phase, immune checkpoint genes (e.g., CTLA4, EOMES, TIGIT, LGALS3 ) were predominantly expressed by infiltrating T lymphocytes, implying that T cell exhaustion drives inflammation. Moreover, chemokine genes strongly correlated with ALT levels, such as CCL20 and CXCL8 , were primarily expressed by macrophages, suggesting that macrophages secrete chemokines to recruit immune‐exhausted T lymphocytes into liver tissues. (F) Workflow for clinical histopathology validation: Liver slides from 12 CHB patients in the hepatitis stage and five CHB patients in the infection phase were analyzed. Multiplex immunostaining was performed on eight sequential slides using the parameters outlined in the blue box. These images were created using Biorender ( https://biorender.com/ ). ALT, alanine aminotransferase; CHB, chronic hepatitis B; HBeAg, hepatitis B e‐antigen; PSEA, population‐specific expression analysis.

Article Snippet: Primary antibodies comprising CD3 (abcam; ab16669), CD20 (abcam; ab78237), CD68 (abcam; ab213363), NKp46 (abcam; ab224703), TIGIT (abcam; ab243903), CTLA4 (CST; #53560S), CXCL8 (CST; #94407S), CCL20 (abcam; ab224188), IFI44 (abcam; ab236657), IFI16 (abcam; ab169788), ISG15 (abcam; ab285367), ISG20 (abcam; ab157477), HBcAg (abcam; ab8639), and DAPI staining solution were bought from Abcam Company (Figures , , ).

Techniques: Gene Expression, Infection, Expressing, Histopathology, Biomarker Discovery, Multiplex Assay, Immunostaining

Journal: iMeta

Article Title: Cell‐type‐specific expression analysis of liver transcriptomics with clinical parameters to decipher the cause of intrahepatic inflammation in chronic hepatitis B

doi: 10.1002/imt2.221

Figure Lengend Snippet: Pathological validation of markers indicative of T cell exhaustion and chemokines of macrophage activation . (A) Double immunohistochemical staining of CTLA4 with CD3 in liver specimens of HBeAg‐positive hepatitis patients and HBeAg‐positive infection patients. (B) Column diagram of CTLA4+ and CD3+CTLA4+ cells for CHB hepatitis, CHB infection, and infiltrated site in CHB hepatitis slides. (C) Double immunohistochemical staining of TIGIT with CD3 in liver specimens of HBeAg‐positive hepatitis patients and HBeAg‐positive infection patients. (D) Column diagram of TIGIT+ and CD3+TIGIT+ cells for CHB hepatitis, CHB infection, and infiltrated site in CHB hepatitis slides. (E) Double labeling of CCL20 (red) with CD68 (green) and CD3 (green) in liver specimens from three HBeAg‐positive hepatitis patients. (F) Column diagram of CCL20+, CD3+CCL20+, and CD68+CCL20+ cells for CHB hepatitis, CHB infection, and infiltrated site in CHB hepatitis slides. (G) Double labeling of CXCL8 (red) with CD68 (green) and CD3 (green) in liver specimens from three HBeAg‐positive hepatitis patients. (H) Column diagram of CXCL8+, CD3+CXCL8+, and CD68+CXCL8+ cells for CHB hepatitis, CHB infection, and infiltrated site in CHB hepatitis slides. CTLA4 and TIGIT were colocalized with CD3 at the inflammatory site, suggesting that exhausted CD3+ T cells were widely infiltrated into the liver at these sites during hepatitis. The release of chemokines such as CCL20 and CXCL8 was primarily from CD68+ macrophages and highly expressed in the hepatitis group and inflammatory sites infiltrated by CD3+ T cells, contributing to intrahepatic inflammation. CHB, chronic hepatitis B; HBeAg, hepatitis B e‐antigen.

Article Snippet: Primary antibodies comprising CD3 (abcam; ab16669), CD20 (abcam; ab78237), CD68 (abcam; ab213363), NKp46 (abcam; ab224703), TIGIT (abcam; ab243903), CTLA4 (CST; #53560S), CXCL8 (CST; #94407S), CCL20 (abcam; ab224188), IFI44 (abcam; ab236657), IFI16 (abcam; ab169788), ISG15 (abcam; ab285367), ISG20 (abcam; ab157477), HBcAg (abcam; ab8639), and DAPI staining solution were bought from Abcam Company (Figures , , ).

Techniques: Biomarker Discovery, Activation Assay, Immunohistochemical staining, Staining, Infection, Labeling

Journal: Cell reports

Article Title: Differences in syncytia formation by SARS-CoV-2 variants modify host chromatin accessibility and cellular senescence via TP53

doi: 10.1016/j.celrep.2023.113478

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Primary antibodies used were anti-FLAG M2 (Sigma-Aldrich, F1804, 1:2000), anti-DYKDDDDK (FLAG) Tag (CST, 14793S, 1:1000), anti-ACE2 (ProSci, 3217, 1:1000), anti-ACTB (Sigma, A5316, 1:5000), anti-SARS-CoV-2-Spike (Sino Biological, 40592-MM117, 1:2000), anti-MERS-CoV-Spike (Sino Biological, 100208-RP02, 1:1000), anti-Renilla Luciferase (abcam, ab185925, 1:1000), anti-VSV-G [8G5F11] (Kerafast, EB0010, 1:1000), anti-VSV-M [23H12] (Kerafast, EB0011, 1:1000), anti-HA (CST, 2367S, 1:1000), anti-TP53 (CST, 9282S, 1:1000), anti-CDKN1A/p21 (CST, 2947S, 1:1000), anti-CXCL8/IL8 [E5F5Q] (CST, 94407, 1:1000), anti-CXCL1/2 [E5M6D] (CST, 24376, 1:1000), anti-Lamin B1 (abcam, ab16048, 1:1000), anti-GAPDH [14C10] (CST, 2118, 1:1000), and anti-Strep II Tag (Novus, NBP2-43735, 1:1000).

Techniques: FLAG-tag, Luciferase, Virus, Recombinant, Reverse Transcription, SYBR Green Assay, Activity Assay, Software, CRISPR