Journal: Frontiers in Oncology

Article Title: PET/CT Imaging of Activated Cancer-Associated Fibroblasts Predict Response to PD-1 Blockade in Gastric Cancer Patients

doi: 10.3389/fonc.2021.802257

Figure Lengend Snippet: Association between the expression of FAP and immunosuppressive TME. (A) Representative images of IHC staining of FAP, macrophage M2 (CD163+), MDSC (CD11b+/CD33+) infiltration, and PD-1 expression in GC tissue microarrays. Scale bar = 100 µm. (B) Quantitative results of IHC for CD163, CD11b, CD33, and PD-1 in groups with low intensity and high intensity of FAP in GC tissue microarrays. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The slides and GC tissue microarrays ( n = 31) were stained with rabbit anti-FAP (Abcam, ab53066; Cambridge, MA, USA), anti-CD11b (Abcam, ab133357), anti-CD33 (Abcam, ab269456), anti-CD163(Abcam, ab182422), and anti-PD-1 (Abcam, ab243644) primary antibodies overnight at 4°C and incubated with anti-rabbit secondary antibody for 1 h at room temperature.

Techniques: Expressing, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: PET/CT Imaging of Activated Cancer-Associated Fibroblasts Predict Response to PD-1 Blockade in Gastric Cancer Patients

doi: 10.3389/fonc.2021.802257

Figure Lengend Snippet: FAPI imaging characterizes the immunosuppressive TME in GC. (A) Representative 68 Ga-FAPI-04 PET/CT images of patients with gastric cancer and IHC staining images for tumor specimens from gastroscopical biopsy of FAP, MDSC (CD11b+/CD33+), and macrophage M2 (CD163+) infiltration. Scale bar = 100 µm. The primary lesions are marked by red arrows, and the metastatic lesions are marked by white arrows. (B) Correlation analysis of TMEscore and the corresponding mean 68 Ga-FAPI-04 uptake ratios in primary gastric cancer or liver metastases.

Article Snippet: The slides and GC tissue microarrays ( n = 31) were stained with rabbit anti-FAP (Abcam, ab53066; Cambridge, MA, USA), anti-CD11b (Abcam, ab133357), anti-CD33 (Abcam, ab269456), anti-CD163(Abcam, ab182422), and anti-PD-1 (Abcam, ab243644) primary antibodies overnight at 4°C and incubated with anti-rabbit secondary antibody for 1 h at room temperature.

Techniques: Imaging, Positron Emission Tomography-Computed Tomography, Immunohistochemistry

Journal: Cancer Cell International

Article Title: Overexpression of microRNA-367 inhibits angiogenesis in ovarian cancer by downregulating the expression of LPA1

doi: 10.1186/s12935-020-01551-x

Figure Lengend Snippet: Primer sequences for RT-qPCR

Article Snippet: The membrane was then incubated overnight at 4 °C with diluted primary antibody, i.e. one of the rabbit antibodies LPA1 (R&D system, Minneapolis, MN, USA, AF9963, 1:2000), MCM2 (R&D system, AF5778, 1:2000), MMP2 (R&D system, AF902, 1:2000), MMP9 (R&D system, AF909, 1:2000), VEGF (R&D system, AF-493-NA, 1:2000), and GAPDH (R&D system, AF5718, 1:2000).

Techniques:

Journal: Cancer Cell International

Article Title: Overexpression of microRNA-367 inhibits angiogenesis in ovarian cancer by downregulating the expression of LPA1

doi: 10.1186/s12935-020-01551-x

Figure Lengend Snippet: miR-367 is poorly expressed but LPA1 is highly expressed in ovarian cancer cells. a Differential expression of miR-367 in cancer tissue samples and adjacent normal tissue samples in ovarian cancer microarray dataset GSE48485. b Binding site map of miR-367 to the target gene LPA1 in the StarBase database. c Box diagram of differential expression of LPA1 gene in cancer tissue samples and adjacent normal tissue samples in ovarian cancer microarray dataset GSE66957

Article Snippet: The membrane was then incubated overnight at 4 °C with diluted primary antibody, i.e. one of the rabbit antibodies LPA1 (R&D system, Minneapolis, MN, USA, AF9963, 1:2000), MCM2 (R&D system, AF5778, 1:2000), MMP2 (R&D system, AF902, 1:2000), MMP9 (R&D system, AF909, 1:2000), VEGF (R&D system, AF-493-NA, 1:2000), and GAPDH (R&D system, AF5718, 1:2000).

Techniques: Quantitative Proteomics, Microarray, Binding Assay

Journal: Cancer Cell International

Article Title: Overexpression of microRNA-367 inhibits angiogenesis in ovarian cancer by downregulating the expression of LPA1

doi: 10.1186/s12935-020-01551-x

Figure Lengend Snippet: miR-367 downregulates LPA1 expression. a Bioinformatics prediction of binding sites for miR-367 and LPA1. b Luminescence activity of LPA1 by dual luciferase reporter gene assay. c RIP assay detection of the binding of miR-367 and LPA1 to Ago2. d Detection of relative expression of LPA1 in cancer tissues and adjacent normal tissues by RT-qPCR. e Analysis of protein level of LPA1 in cancer and adjacent normal tissues by western blot analysis. f Statistical expression of protein level of LPA1. g Correlation analysis between miR-367 and LPA1. h RT-qPCR results of the relative expression level of LPA1 in each group of cells. i Western blot analysis of LPA1 protein levels in each group of cells. j Statistical protein expression level of LPA1. Measurement data were expressed as mean ± standard deviation. Data in compliance with normal distribution and homogeneity between two groups were compared using t test. Comparisons among multiple groups were conducted by one-way ANOVA followed by Tukey’s post hoc test. * p < 0.05 vs. normal tissues or cells; # p < 0.05 vs. Inhibitor NC The experiment was repeated three times independently

Article Snippet: The membrane was then incubated overnight at 4 °C with diluted primary antibody, i.e. one of the rabbit antibodies LPA1 (R&D system, Minneapolis, MN, USA, AF9963, 1:2000), MCM2 (R&D system, AF5778, 1:2000), MMP2 (R&D system, AF902, 1:2000), MMP9 (R&D system, AF909, 1:2000), VEGF (R&D system, AF-493-NA, 1:2000), and GAPDH (R&D system, AF5718, 1:2000).

Techniques: Expressing, Binding Assay, Activity Assay, Luciferase, Reporter Gene Assay, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: Cancer Cell International

Article Title: Overexpression of microRNA-367 inhibits angiogenesis in ovarian cancer by downregulating the expression of LPA1

doi: 10.1186/s12935-020-01551-x

Figure Lengend Snippet: Low expression of miR-367 upregulates the expression of LPA1 to promote proliferation, invasion, and angiogenesis of ovarian cancer A2780 cells. a RT-qPCR analysis results of the efficiency of LPA1 silencing by appropriate siRNAs. b miR-367 and LPA1 expression in cells determined by using RT-qPCR. c A2780 cell proliferation ability detected by Transwell assay (scale bar = 25 μm). d Invasion ability of A2780 cells measured by Transwell assay (scale bar = 50 μm). e Relative length of blood vessels. f Relative number of blood vessel branches. g – j Statistics of CAM blood vessel number, area, vessel area ratio and tissues area. k RT-qPCR detection of mRNA levels of related factors. l Western blot analysis of protein levels of related factors. m Statistics of protein expression levels of related factors. Measurement data were expressed as mean ± standard deviation. Data in compliance with normal distribution and homogeneity among multiple groups were conducted by one-way ANOVA followed by Tukey’s post hoc test. * p < 0.05 vs. NC. # p < 0.05 vs. cells treated with si-LPA1. The experiment was repeated three times independently

Article Snippet: The membrane was then incubated overnight at 4 °C with diluted primary antibody, i.e. one of the rabbit antibodies LPA1 (R&D system, Minneapolis, MN, USA, AF9963, 1:2000), MCM2 (R&D system, AF5778, 1:2000), MMP2 (R&D system, AF902, 1:2000), MMP9 (R&D system, AF909, 1:2000), VEGF (R&D system, AF-493-NA, 1:2000), and GAPDH (R&D system, AF5718, 1:2000).

Techniques: Expressing, Quantitative RT-PCR, Transwell Assay, Western Blot, Standard Deviation

Journal: Cancer Cell International

Article Title: Overexpression of microRNA-367 inhibits angiogenesis in ovarian cancer by downregulating the expression of LPA1

doi: 10.1186/s12935-020-01551-x

Figure Lengend Snippet: Low expression of LPA1 inhibits tumorigenic ability of ovarian cancer in nude mice. a Volume of mice tumor. b Weight of mice tumor. c RT-qPCR detection results of mRNA levels of related factors in tumor tissues. d Western blot analysis of the protein levels of related factors in tumor tissues. e Statistical graph of protein levels of related factors. f Immunohistochemistry for detecting CD34 expression (200×). Measurement data were expressed as mean ± standard deviation. Data in compliance with normal distribution and homogeneity between two groups were compared using t test. Comparisons among multiple groups were conducted by one-way ANOVA followed by Tukey’s post hoc test. Statistics of tumor volume at different time points was analyzed using repeated measures ANOVA. * p < 0.05 vs. normal tissues or cells. The experiment was repeated three times independently

Article Snippet: The membrane was then incubated overnight at 4 °C with diluted primary antibody, i.e. one of the rabbit antibodies LPA1 (R&D system, Minneapolis, MN, USA, AF9963, 1:2000), MCM2 (R&D system, AF5778, 1:2000), MMP2 (R&D system, AF902, 1:2000), MMP9 (R&D system, AF909, 1:2000), VEGF (R&D system, AF-493-NA, 1:2000), and GAPDH (R&D system, AF5718, 1:2000).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Standard Deviation

Journal: Cancer Cell International

Article Title: Overexpression of microRNA-367 inhibits angiogenesis in ovarian cancer by downregulating the expression of LPA1

doi: 10.1186/s12935-020-01551-x

Figure Lengend Snippet: The regulatory mechanism map. Overexpressed miR-367 could reduce the expression of LPA1 and inhibit the proliferation and invasion of cancer cells, and the process of tumor-induced angiogenesis, hereby repressing ovarian cancer development

Article Snippet: The membrane was then incubated overnight at 4 °C with diluted primary antibody, i.e. one of the rabbit antibodies LPA1 (R&D system, Minneapolis, MN, USA, AF9963, 1:2000), MCM2 (R&D system, AF5778, 1:2000), MMP2 (R&D system, AF902, 1:2000), MMP9 (R&D system, AF909, 1:2000), VEGF (R&D system, AF-493-NA, 1:2000), and GAPDH (R&D system, AF5718, 1:2000).

Techniques: Expressing

Journal: Cancer research

Article Title: Myeloid-derived suppressor cells endow stem-like qualities to breast cancer cells through IL-6/STAT3 and NO/NOTCH cross-talk signaling

doi: 10.1158/0008-5472.CAN-15-2528

Figure Lengend Snippet: (a–d) Phenotype of MDSCs. Fresh breast cancer tissues were separated into single cell suspension. The cells were stained with isotype control antibodies and relevant antibodies. Polychromatic flow cytometry analysis was performed on these cells by gating on single cells in different gates (G1–G3). (a) The relationship among CD45+, CD5+, CD19+ and CD33+ cells. Upper panels: FSC, SSC, and CD45 gates; Middle panels: isotype controls in gates 1*2*3. Lower panels: CD19, CD5 and CD33 in gates 1*2*3; (b) The phenotype of MDSCs. The expression of CD11b, CD14, CD15, CD16, HLA-DR and 7-AAD was analyzed in CD5−CD19−CD33+CD45+ cells. (c) The percentage of different antigen expressing cells in CD5−CD19−CD33+CD45+ cells. (d) The percentage of CD33 expressing cells in CD45+ cells. One of 10 representative patients is shown (a, b). Columns represent the mean ± SEM (c, d) (n = 10).

Article Snippet: Tissues were stained with polyclonal rabbit anti-human-CD33 (1/10 dilution, DAKO) ( 13 ), or mouse anti-human ALDH1 (Becton Dickinson) ( 24 ).

Techniques: Staining, Flow Cytometry, Expressing

Journal: Cancer research

Article Title: Myeloid-derived suppressor cells endow stem-like qualities to breast cancer cells through IL-6/STAT3 and NO/NOTCH cross-talk signaling

doi: 10.1158/0008-5472.CAN-15-2528

Figure Lengend Snippet: Immunohistochemistry was performed in human breast tumor tissues. CD33+ cells were quantified in 0.6mm2.

Article Snippet: Tissues were stained with polyclonal rabbit anti-human-CD33 (1/10 dilution, DAKO) ( 13 ), or mouse anti-human ALDH1 (Becton Dickinson) ( 24 ).

Techniques: Immunohistochemistry