Journal: Journal of Dental Sciences

Article Title: Serpin peptidase inhibitor, clade E, member 2 is associated with malignant progression and clinical prognosis in oral squamous cell carcinoma

doi: 10.1016/j.jds.2023.05.024

Figure Lengend Snippet: mRNA expression with SERPINE2 diagnostic and prognostic values. (A) SERPINE2 mRNA expression levels in 546 normal, tumor, and metastatic head and neck squamous cell carcinoma (HNSCC) tissues. SERPINE2 expressions were higher in tumor tissues than in normal tissues in TNMplot ( P = 2.009e-07). (B) High SERPINE2 mRNA expression was significantly correlated with poor 5-year overall survival in KMPlot ( P = 0.014). (C) Heat map and boxplot of SERPINE2 mRNA expression in 40 OSCC tissue pairs (GSE37991). SERPINE2 is upregulated in human OSCC. Red, upregulated; green, downregulated. Array intensity of SERPINE2 in 40 OSCC tumors compared with their adjacent normal tissues; SERPINE2 intensity is expressed as the log2 ratios. Data are represented as mean ± standard deviation; ∗∗∗∗, P < 0.0001.

Article Snippet: The sections were sequentially incubated at room temperature with monoclonal mouse anti-human SERPINE2 antibody (1:100; #MA5-25936; Invitrogen, Carlsbad, CA, USA) for 60 min, biotinylated secondary antibody for 10 min, and peroxidase-conjugated streptavidin for 10 min.

Techniques: Expressing, Diagnostic Assay, Standard Deviation

Journal: Journal of Dental Sciences

Article Title: Serpin peptidase inhibitor, clade E, member 2 is associated with malignant progression and clinical prognosis in oral squamous cell carcinoma

doi: 10.1016/j.jds.2023.05.024

Figure Lengend Snippet: Representative sections of non-cancer epithelia and OSCC were stained with hematoxylin and eosin and immunostained for SERPINE2. (A) Non-cancer epithelia, (B) weak (1+), (C) moderate (2+) and (D) strong (3+). Scale bar in (A), 100 μm. The scale bar applies to all panels.

Article Snippet: The sections were sequentially incubated at room temperature with monoclonal mouse anti-human SERPINE2 antibody (1:100; #MA5-25936; Invitrogen, Carlsbad, CA, USA) for 60 min, biotinylated secondary antibody for 10 min, and peroxidase-conjugated streptavidin for 10 min.

Techniques: Staining

Journal: Journal of Dental Sciences

Article Title: Serpin peptidase inhibitor, clade E, member 2 is associated with malignant progression and clinical prognosis in oral squamous cell carcinoma

doi: 10.1016/j.jds.2023.05.024

Figure Lengend Snippet: Relationships between the immunoscore of SERPINE2 and clinicopathological parameters in 122 patients.

Article Snippet: The sections were sequentially incubated at room temperature with monoclonal mouse anti-human SERPINE2 antibody (1:100; #MA5-25936; Invitrogen, Carlsbad, CA, USA) for 60 min, biotinylated secondary antibody for 10 min, and peroxidase-conjugated streptavidin for 10 min.

Techniques:

Journal: Journal of Dental Sciences

Article Title: Serpin peptidase inhibitor, clade E, member 2 is associated with malignant progression and clinical prognosis in oral squamous cell carcinoma

doi: 10.1016/j.jds.2023.05.024

Figure Lengend Snippet: High SERPINE2 expression was correlated with worse overall survival rates. Kaplan–Meier survival curve in 122 OSCC patients.

Article Snippet: The sections were sequentially incubated at room temperature with monoclonal mouse anti-human SERPINE2 antibody (1:100; #MA5-25936; Invitrogen, Carlsbad, CA, USA) for 60 min, biotinylated secondary antibody for 10 min, and peroxidase-conjugated streptavidin for 10 min.

Techniques: Expressing

Journal: Journal of Dental Sciences

Article Title: Serpin peptidase inhibitor, clade E, member 2 is associated with malignant progression and clinical prognosis in oral squamous cell carcinoma

doi: 10.1016/j.jds.2023.05.024

Figure Lengend Snippet: Univariate and multivariate analysis of SERPINE2 in 122 oral squamous cell carcinoma patients.

Article Snippet: The sections were sequentially incubated at room temperature with monoclonal mouse anti-human SERPINE2 antibody (1:100; #MA5-25936; Invitrogen, Carlsbad, CA, USA) for 60 min, biotinylated secondary antibody for 10 min, and peroxidase-conjugated streptavidin for 10 min.

Techniques: Expressing

Journal: Journal of Dental Sciences

Article Title: Serpin peptidase inhibitor, clade E, member 2 is associated with malignant progression and clinical prognosis in oral squamous cell carcinoma

doi: 10.1016/j.jds.2023.05.024

Figure Lengend Snippet: SERPINE2 knockdown reduced cell proliferation and invasion in HSC3 and Cal27 cells. (A) SERPINE2 expression levels in OSCC cell lines were evaluated using western blotting. (B) Immunoblotting analysis of SERPINE2 in HSC3 and Cal27 cells transduced with SERPINE2 shRNA and control shRNA (shLuc). (C) Growth curves of HSC3 cells with SERPINE2 knockdown. The migration and invasion of HSC3 cells transduced with SERPINE2 shRNA or control vector were assessed using Transwell assays. (D) Growth curves of Cal27 cells with SERPINE2 knockdown. Transwell assays were performed to analyze the migration and invasion of Cal27 cells transduced with SERPINE2 shRNA or a control vector. bar: SEM; ∗ P < 0.05; ∗∗ P < 0.001.

Article Snippet: The sections were sequentially incubated at room temperature with monoclonal mouse anti-human SERPINE2 antibody (1:100; #MA5-25936; Invitrogen, Carlsbad, CA, USA) for 60 min, biotinylated secondary antibody for 10 min, and peroxidase-conjugated streptavidin for 10 min.

Techniques: Expressing, Western Blot, Transduction, shRNA, Migration, Plasmid Preparation

Journal: Oncogene

Article Title: A slow-cycling subpopulation of melanoma cells with highly invasive properties.

doi: 10.1038/onc.2017.341

Figure Lengend Snippet: Figure 3. Label-retaining, disseminated melanoma cells express high levels of SerpinE2. (a) Table lists proteins identified after whole-cell proteomic analyses of LRC and non-LRC. The last column lists the cell lines with differential expression of the indicated protein. (b) Violin plots (left panels) illustrate quantification of SerpinE2 (white spot, top) and BMP1 (white spots, bottom) as determined by mRNA FISH. mRNA counts for single cells are shown on the Y-axes. Note differences in Y-axes. Representative photomicrographs are shown on the right (total magnification: 100 × ). (c) Protein expression was analyzed by immunofluorescence and quantified at single-cell levels (scatter plots on left and representative images on right). LRC (red), non-LRC (orange), total cells (gray). (d) Flow cytometry of disseminated WM989 melanoma cells detected in three mouse organs (top panel). The percentage of cells double positive for CD146 and SerpinE2 is shown in the upper right quadrants; the lower right quadrants indicate the percentage of cells singly positive for CD146. Micrographs show examples of disseminated SerpinE2-positive cells (arrows, middle and right panels) in mouse lungs. Left panel: negative control.

Article Snippet: Formalin-fixed, paraffin-embedded tissue sections (1– 2 μm) were stained with mouse anti-human SerpinE2 antibody (Origene) followed by polymer staining and Fast-Red for 7 min at RT.

Techniques: Quantitative Proteomics, Expressing, Flow Cytometry, Negative Control

Journal: Oncogene

Article Title: A slow-cycling subpopulation of melanoma cells with highly invasive properties.

doi: 10.1038/onc.2017.341

Figure Lengend Snippet: Figure 5. SerpinE2 expression is increased in malignant cells and correlates with tumor progression. (a) Western blot analysis of SerpinE2 secretion by melanoma cells (n = 21) and human melanocytes (n = 5) into culture supernatants. Recombinant SerpinE2 protein (500 ng) was used as a positive control and Ponceau’s staining is shown for loading control. Graph shows quantification (right panel). (b) SerpinE2 staining of 3D skin reconstructs with melanocytes (left) and melanoma cells (right). (c) GEM_1375 data set analysis of SerpinE2 mRNA expression in melanomas, non-malignant nevi and normal skin. (d) Examples of immune histochemistry at two different magnifications of: normal skin, benign nevi, in situ, vertical growth phase (VGP) and lymph node metastatic melanomas (n = 15 for each group). SerpinE2 protein expression is indicated by purple staining. Arrows indicate the dotted SerpinE2 expression pattern found in benign nevi only. (e) Intensity score of SerpinE2 expression in normal skin, benign nevi and malignant melanomas (radial growth phase (RGP), VGP and metastatic). Bars represent mean ± SEM of tissue sections of 15 specimens from each group; P-value after t-student test are given.

Article Snippet: Formalin-fixed, paraffin-embedded tissue sections (1– 2 μm) were stained with mouse anti-human SerpinE2 antibody (Origene) followed by polymer staining and Fast-Red for 7 min at RT.

Techniques: Expressing, Western Blot, Recombinant, Positive Control, Staining, Control, In Situ

Journal: Oncogene

Article Title: A slow-cycling subpopulation of melanoma cells with highly invasive properties

doi: 10.1038/onc.2017.341

Figure Lengend Snippet: Label-retaining, disseminated melanoma cells express high levels of SerpinE2. ( a ) Table lists proteins identified after whole-cell proteomic analyses of LRC and non-LRC. The last column lists the cell lines with differential expression of the indicated protein. ( b ) Violin plots (left panels) illustrate quantification of SerpinE2 (white spot, top) and BMP1 (white spots, bottom) as determined by mRNA FISH. mRNA counts for single cells are shown on the Y -axes. Note differences in Y -axes. Representative photomicrographs are shown on the right (total magnification: 100 ×). ( c ) Protein expression was analyzed by immunofluorescence and quantified at single-cell levels (scatter plots on left and representative images on right). LRC (red), non-LRC (orange), total cells (gray). ( d ) Flow cytometry of disseminated WM989 melanoma cells detected in three mouse organs (top panel). The percentage of cells double positive for CD146 and SerpinE2 is shown in the upper right quadrants; the lower right quadrants indicate the percentage of cells singly positive for CD146. Micrographs show examples of disseminated SerpinE2-positive cells (arrows, middle and right panels) in mouse lungs. Left panel: negative control.

Article Snippet: Samples were boiled 10 min in NuPAGE LDS sample buffer (Thermo Fisher Scientific), loaded on 10% gels, transferred onto nitrocellulose filters and immunoblotted with mouse anti-human SerpinE2 antibody (R&D), diluted 1:500.

Techniques: Quantitative Proteomics, Expressing, Immunofluorescence, Flow Cytometry, Negative Control

Journal: Oncogene

Article Title: A slow-cycling subpopulation of melanoma cells with highly invasive properties

doi: 10.1038/onc.2017.341

Figure Lengend Snippet: SerpinE2 drives melanoma invasiveness. ( a ) Boyden chamber invasion assay of sorted label-retaining cells (LRC) or non-LRC in presence (blue columns) or absence (gray columns) of human recombinant SerpinE2. Bars show fold change in invasion (mean±s.e.m.) of samples with SerpinE2 over untreated cells. ( b ) Invasion assay in presence of an anti-human SerpinE2 neutralizing antibody or Isotype control (ctrl). The percentage of invading cells after neutralization is shown. Data represent mean±s.e.m. of three independent experiments. ( c ) Invasion assay after SerpinE2 knockdown. Data shown are percentage of invading cells found after silencing with 5 different shRNA (Sh_13-17) relative to Sh_ctrl (ctrl). Bars represent mean±s.e.m.

Article Snippet: Samples were boiled 10 min in NuPAGE LDS sample buffer (Thermo Fisher Scientific), loaded on 10% gels, transferred onto nitrocellulose filters and immunoblotted with mouse anti-human SerpinE2 antibody (R&D), diluted 1:500.

Techniques: Invasion Assay, Recombinant, Control, Neutralization, Knockdown, shRNA

Journal: Oncogene

Article Title: A slow-cycling subpopulation of melanoma cells with highly invasive properties

doi: 10.1038/onc.2017.341

Figure Lengend Snippet: SerpinE2 expression is increased in malignant cells and correlates with tumor progression. ( a ) Western blot analysis of SerpinE2 secretion by melanoma cells ( n =21) and human melanocytes ( n =5) into culture supernatants. Recombinant SerpinE2 protein (500 ng) was used as a positive control and Ponceau’s staining is shown for loading control. Graph shows quantification (right panel). ( b ) SerpinE2 staining of 3D skin reconstructs with melanocytes (left) and melanoma cells (right). ( c ) GEM_1375 data set analysis of SerpinE2 mRNA expression in melanomas, non-malignant nevi and normal skin. ( d ) Examples of immune histochemistry at two different magnifications of: normal skin, benign nevi, in situ , vertical growth phase (VGP) and lymph node metastatic melanomas ( n =15 for each group). SerpinE2 protein expression is indicated by purple staining. Arrows indicate the dotted SerpinE2 expression pattern found in benign nevi only. ( e ) Intensity score of SerpinE2 expression in normal skin, benign nevi and malignant melanomas (radial growth phase (RGP), VGP and metastatic). Bars represent mean±SEM of tissue sections of 15 specimens from each group; P- value after t -student test are given.

Article Snippet: Samples were boiled 10 min in NuPAGE LDS sample buffer (Thermo Fisher Scientific), loaded on 10% gels, transferred onto nitrocellulose filters and immunoblotted with mouse anti-human SerpinE2 antibody (R&D), diluted 1:500.

Techniques: Expressing, Western Blot, Recombinant, Positive Control, Staining, Control, In Situ