Journal: Molecular Cancer

Article Title: Exosomal miR-4745-5p/3911 from N2-polarized tumor-associated neutrophils promotes gastric cancer metastasis by regulating SLIT2

doi: 10.1186/s12943-024-02116-6

Figure Lengend Snippet: SLIT2 is a common target for miR-4745-5p and miR-3911 in gastric cancer cells. ( A ) Bioinformatic analysis of the common target genes of miR-4745-5p and miR-3911. ( B-C ) qRT-PCR analyses of SLIT2 gene expression in miR-4745-5p and miR-3911 overexpressing ( B ) and knockdown ( C ) gastric cancer cells. ( D-E ) Western blot assays for SLIT2 protein expression in miR-4745-5p and miR-3911 overexpressing and knockdown gastric cancer cells. ( F ) The potential binding sites of miR-4745-5p and miR-3911 with SLIT2. (G-J) Luciferase reporter assays for SLIT2 reporter gene in miR-4745-5p and miR-3911 overexpressing ( G-H ) and knockdown ( I-J ) gastric cancer cells. * P < 0.05 and ** P < 0.01; ns, no significant change

Article Snippet: A recent study that uses in vivo genome-wide CRISPR knockout identifies SLIT2 knockout circulating tumor cells (CTCs) are highly metastatic [ ].

Techniques: Quantitative RT-PCR, Gene Expression, Knockdown, Western Blot, Expressing, Binding Assay, Luciferase

Journal: Molecular Cancer

Article Title: Exosomal miR-4745-5p/3911 from N2-polarized tumor-associated neutrophils promotes gastric cancer metastasis by regulating SLIT2

doi: 10.1186/s12943-024-02116-6

Figure Lengend Snippet: N2-polarized neutrophils derived exosomes promote gastric cancer metastasis via regulation of SLIT2. ( A ) qRT-PCR analyses of SLIT2 gene expression in control and N2-Exo treated gastric cancer cells. ( B ) Western blot assays for SLIT2 protein expression in control and N2-Exo treated gastric cancer cells. ( C-D ) Western blot assays for SLIT2 protein expression in control and N2-Exo treated gastric cancer cells with or without miR-4745-5p and miR-3911 knockdown. ( E ) Luciferase reporter assays for N2-Exo-treated gastric cancer cells transfected with wild-type and mutant SLIT2 reporter gene vector. ( F-H ) Transwell migration and matrigel invasion assays for gastric cancer cells (HGC27) treated with N2-Exo and transfected with SLIT2 (Ad-SLIT2). Scale bars, 100 μm. ( I ) The signal intensities of bioluminescent imaging in the lung tissues of indicated groups. ( J ) Lung tissues with metastatic nodules in the indicated groups. ( K ) H&E staining of lung tissues in the indicated groups. Top panel, scale bars, 1 mm. Bottom panel, scale bars, 100 μm. ( L ) The number of metastatic nodules in the lung tissues of indicated groups. ( M ) SLIT2 staining in metastatic lung tissues. Scale bars, 100 μm. * P < 0.05 and ** P < 0.01

Article Snippet: A recent study that uses in vivo genome-wide CRISPR knockout identifies SLIT2 knockout circulating tumor cells (CTCs) are highly metastatic [ ].

Techniques: Derivative Assay, Quantitative RT-PCR, Gene Expression, Control, Western Blot, Expressing, Knockdown, Luciferase, Transfection, Mutagenesis, Plasmid Preparation, Migration, Imaging, Staining

Journal: Molecular Cancer

Article Title: Exosomal miR-4745-5p/3911 from N2-polarized tumor-associated neutrophils promotes gastric cancer metastasis by regulating SLIT2

doi: 10.1186/s12943-024-02116-6

Figure Lengend Snippet: Circulating levels of miR-4745-5p and miR-3911 are elevated in N2-polarized neutrophils derived exosomes from gastric cancer patients. ( A-B ) Immunohistochemical staining of CD66 and SLIT2 in gastric cancer tissues with or without metastasis ( n = 25). Scale bars, 100 μm. ( C-D ) qRT-PCR analyses of miR-4745-5p ( C ) and miR-3911 ( D ) expression in serum samples from healthy controls (HC), benign gastric diseases (BGD), and gastric cancer (GC) patients ( n = 30). ( E-F ) The area under curve (AUC) to distinguish GC from GC ( E ) and BGD ( F ) groups. ( G-J ) The expression of miR-4745-5p ( G ) and miR-3911 ( I ) in serum samples ( n = 30) from HC, BGD, GC patients detected by droplet digital PCR (ddPCR) and the statistical analysis result ( H and J ). ( K-L ) The area under curve (AUC) to distinguish GC from GC ( K ) and BGD ( L ) groups for ddPCR analyses of miR-4745-5p and miR-3911 expression in serum neutrophil exosomes. * P < 0.05, ** P < 0.01 and *** P < 0.001. All samples are providing from our own source of patient samples

Article Snippet: A recent study that uses in vivo genome-wide CRISPR knockout identifies SLIT2 knockout circulating tumor cells (CTCs) are highly metastatic [ ].

Techniques: Derivative Assay, Immunohistochemical staining, Staining, Quantitative RT-PCR, Expressing, Digital PCR

Journal: Molecular Cancer

Article Title: Exosomal miR-4745-5p/3911 from N2-polarized tumor-associated neutrophils promotes gastric cancer metastasis by regulating SLIT2

doi: 10.1186/s12943-024-02116-6

Figure Lengend Snippet: Proposed model of the role of exosomes-mediated crosstalk between N2 TANs and gastric cancer cells in gastric cancer metastasis. Gastric cancer cells induce the N2 polarization of neutrophils via a glycolysis-dependent manner. In turn, N2-polarized neutrophils promote gastric cancer metastasis through exosomal miR-4745-5p/3911-mediated inhibition of SLIT2 expression in gastric cancer cells. Serum neutrophil exosomal miR-4745-5p/3911 represents a novel biomarker for GC diagnosis

Article Snippet: A recent study that uses in vivo genome-wide CRISPR knockout identifies SLIT2 knockout circulating tumor cells (CTCs) are highly metastatic [ ].

Techniques: Inhibition, Expressing, Biomarker Discovery

Journal: Current Issues in Molecular Biology

Article Title: Molecular Profiling of Circulating Tumour Cells and Circulating Tumour DNA: Complementary Insights from a Single Blood Sample Utilising the Parsortix ® System

doi: 10.3390/cimb46010050

Figure Lengend Snippet: Publications using the Parsortix ® system: CTC and ctDNA analysis.

Article Snippet: These include the CellSearch ® Circulating Tumor Cell (CTC) Test (Menarini-Silicon Biosystems, Huntingdon Valley, PA, USA): for the enumeration of CTCs of epithelial origin for the monitoring of prognostic information of patients with metastatic breast, colorectal, or prostate cancer, and the Parsortix ® PC1 System (ANGLE plc, Guildford, UK): for the capture and harvest of CTCs from the blood of metastatic breast cancer (MBC) patients for subsequent, user-validated analysis .

Techniques: Methylation, DNA Methylation Assay, Biomarker Discovery, Amplification, Over Expression, Imaging, Immunocytochemistry