Journal: Cancers

Article Title: Reassessing the Role of Tissue Factor Pathway Inhibitor 2 in Neoplastic and Non-Neoplastic Lesions

doi: 10.3390/cancers17091447

Figure Lengend Snippet: Relationship between the regulation of TFPI2 expression in cancer cells and host cells. The signaling pathway constituents numbered 1 to 21 in the figure are described in sections of the text labeled as (1,2), and so forth. Arrows denote activation, while T-shaped symbols represent inhibition. ACTN4, actinin 4; AP-1, activator protein 1; AP-2α, activator protein 2 alpha; CLIP1, CAP-Gly domain-containing linker protein 1 (CLIP1); COX-2, cyclooxygenase-2; DNMT1, DNA methyltransferase 1; EGF, epidermal growth factor; ERK, extracellular signal-regulated kinase; HGF, hepatocyte growth factor; IL-10, interleukin-10; JNK, c-Jun N-terminal kinase; MAPK, mitogen-activated protein kinase; MBD3, methyl-CpG-binding domain protein 3; MMP, matrix metalloproteinase; MYH9, myosin 9; MITF, melanocyte-induced transcription factor; NF-κB, nuclear factor kappa B; PPARγ, peroxisome proliferator-activated receptor gamma; PSAP, prosaposin; SLUG, snail family transcriptional repressor 2; TAM, tumor-associated macrophages; TFPI2, tissue factor pathway inhibitor 2; TGF-β, transforming growth factor beta; TGFBI, transforming growth factor beta-induced; TIRAP, TIR domain containing adaptor protein; TMPRSS4, transmembrane protease serine 4; uPA, urokinase-type plasminogen activator; VEC, vascular endothelial cells; VEGF, vascular endothelial growth factor; VSMC, vascular smooth muscle cells.

Article Snippet: Tosoh Corporation has developed an automated immunoassay for quantifying TFPI2 concentrations in blood, using a one-step immunofluorescence technique with two anti-TFPI2 monoclonal antibodies [ ].

Techniques: Expressing, Labeling, Activation Assay, Inhibition, Binding Assay

Journal: Cancers

Article Title: Reassessing the Role of Tissue Factor Pathway Inhibitor 2 in Neoplastic and Non-Neoplastic Lesions

doi: 10.3390/cancers17091447

Figure Lengend Snippet: Molecules and signaling pathways regulated by TFPI2. The left pie chart illustrates molecules and signaling pathways modulated by TFPI2 that are implicated in the regulation of glucose and lipid metabolism, macrophage polarization, inflammatory responses, immune modulation, and cellular proliferation and survival. In contrast, the right pie chart depicts molecules and signaling cascades associated with extracellular matrix remodeling, cytoskeletal integrity, and cellular motility.

Article Snippet: Tosoh Corporation has developed an automated immunoassay for quantifying TFPI2 concentrations in blood, using a one-step immunofluorescence technique with two anti-TFPI2 monoclonal antibodies [ ].

Techniques: Protein-Protein interactions

Journal: Cancers

Article Title: Reassessing the Role of Tissue Factor Pathway Inhibitor 2 in Neoplastic and Non-Neoplastic Lesions

doi: 10.3390/cancers17091447

Figure Lengend Snippet: Molecular and signaling pathways governing the regulation of TFPI2 expression. The left pie chart depicts factors that promote the upregulation of TFPI2, while the right pie chart illustrates those that facilitate its downregulation. These genes contribute to cancer invasion and metastasis by modulating inflammatory responses, promoting angiogenesis, regulating immune functions, and influencing tumor cell proliferation, survival, and apoptotic pathways.

Article Snippet: Tosoh Corporation has developed an automated immunoassay for quantifying TFPI2 concentrations in blood, using a one-step immunofluorescence technique with two anti-TFPI2 monoclonal antibodies [ ].

Techniques: Protein-Protein interactions, Expressing

Journal: Cancers

Article Title: Reassessing the Role of Tissue Factor Pathway Inhibitor 2 in Neoplastic and Non-Neoplastic Lesions

doi: 10.3390/cancers17091447

Figure Lengend Snippet: The role of TFPI2 in pregnancy-induced hypertension, diabetes, and atherosclerosis. TFPI2 orchestrates extracellular matrix (ECM) remodeling, tissue regeneration, angiogenesis, and immune modulation. The preservation or disturbance of cellular homeostasis through inflammatory processes may result in TFPI2 dysregulation, thereby contributing to the onset and progression of diabetes, atherosclerosis, and pregnancy-induced hypertension. Superscript arrows represent the upregulation of each factor or the activation of pathological or functional processes, whereas subscript arrows denote downregulation or functional inhibition. DNMT1, DNA methyltransferase 1; ERK, extracellular signal-regulated kinase; EZH1, enhancer of zeste 1 polycomb repressive complex 2 subunit; H3K27me3, tri-methylation at lysine 27 of histone H3; IL-1, interleukin-1; MAPK, mitogen-activated protein kinase; MMP, matrix metalloproteinase; MΦ, macrophage; PI3K, phosphatidylinositol-3-kinase; PPARγ, peroxisome proliferator-activated receptor gamma; TFPI2, tissue factor pathway inhibitor 2; TGF-β, transforming growth factor-beta; TNF-α, tumor necrosis factor-alfa; VEC, vascular endothelial cells; VEGFR, vascular endothelial growth factor receptor; VSMC, vascular smooth muscle cells.

Article Snippet: Tosoh Corporation has developed an automated immunoassay for quantifying TFPI2 concentrations in blood, using a one-step immunofluorescence technique with two anti-TFPI2 monoclonal antibodies [ ].

Techniques: Preserving, Activation Assay, Functional Assay, Inhibition, Methylation

Journal: Cancers

Article Title: Reassessing the Role of Tissue Factor Pathway Inhibitor 2 in Neoplastic and Non-Neoplastic Lesions

doi: 10.3390/cancers17091447

Figure Lengend Snippet: Role of TFPI2 in cancer invasion and metastasis. As illustrated in the upper left panel, TFPI2 overexpression suppresses ECM remodeling and inhibits tumor invasion in vitro. Conversely, the upper right panel shows that TFPI2 downregulation facilitates invasion through MMP-mediated ECM degradation. In vivo, TFPI2 influences not only cancer cells but also diverse components of the TME, including inflammatory and immune cells, endothelial cells, cytokines, ECM constituents, and angiogenic factors, thereby playing a pivotal role in modulating tumor malignancy and metastatic dissemination. Superscript arrows represent the upregulation of each factor or the activation of pathological or functional processes, whereas subscript arrows denote downregulation or functional inhibition.

Article Snippet: Tosoh Corporation has developed an automated immunoassay for quantifying TFPI2 concentrations in blood, using a one-step immunofluorescence technique with two anti-TFPI2 monoclonal antibodies [ ].

Techniques: Over Expression, In Vitro, In Vivo, Activation Assay, Functional Assay, Inhibition

Journal: BMC Cancer

Article Title: Tissue factor pathway inhibitor 2 as a serum biomarker for endometrial cancer: a single-center retrospective study

doi: 10.1186/s12885-024-12827-0

Figure Lengend Snippet: Comparison of serum TFPI2 levels between healthy controls and patients with endometrial cancer. The Mann–Whitney U test showed that the serum TFPI2 levels of patients with endometrial cancer were significantly higher than those of healthy controls (median 196.7 pg/mL vs. 83.3 pg/mL). *** indicates p < 0.001. The dotted line indicates 191 pg/mL. TFPI2, tissue factor pathway inhibitor 2

Article Snippet: Immunohistochemistry (IHC) was performed using the same concentration of mouse monoclonal anti-TFPI2 antibody (sc-48380, diluted 1:200, Santa Cruz Biotechnology, Inc., CA, USA) as the primary antibody to ensure consistency of our experimental approach [ ].

Techniques: Comparison, MANN-WHITNEY

Journal: BMC Cancer

Article Title: Tissue factor pathway inhibitor 2 as a serum biomarker for endometrial cancer: a single-center retrospective study

doi: 10.1186/s12885-024-12827-0

Figure Lengend Snippet: Baseline characteristics of 328 patients with endometrial cancer in the TFPI2-positive and TFPI2-negative groups

Article Snippet: Immunohistochemistry (IHC) was performed using the same concentration of mouse monoclonal anti-TFPI2 antibody (sc-48380, diluted 1:200, Santa Cruz Biotechnology, Inc., CA, USA) as the primary antibody to ensure consistency of our experimental approach [ ].

Techniques:

Journal: BMC Cancer

Article Title: Tissue factor pathway inhibitor 2 as a serum biomarker for endometrial cancer: a single-center retrospective study

doi: 10.1186/s12885-024-12827-0

Figure Lengend Snippet: Serum TFPI2 levels of patients with endometrial cancer by histological types and FIGO stages. ( A ) TFPI2 levels were significantly higher in the high-risk group than in the low-risk group (220.8 pg/mL vs. 187.7 pg/mL, p < 0.001). ( B ) TFPI2 levels were significantly higher in stage III and IV than in stage I (stage I, 189.6 pg/mL; stage III, 230.9 pg/mL; stage IV, 312.5 pg/mL). *** indicates p < 0.001 in the Mann–Whitney U test. The dotted line indicates 191 pg/mL. CS, carcinosarcoma; EMG, endometrioid carcinoma grade; FIGO, International Federation of Gynecology and Obstetrics; TFPI2, tissue factor pathway inhibitor 2

Article Snippet: Immunohistochemistry (IHC) was performed using the same concentration of mouse monoclonal anti-TFPI2 antibody (sc-48380, diluted 1:200, Santa Cruz Biotechnology, Inc., CA, USA) as the primary antibody to ensure consistency of our experimental approach [ ].

Techniques: MANN-WHITNEY

Journal: BMC Cancer

Article Title: Tissue factor pathway inhibitor 2 as a serum biomarker for endometrial cancer: a single-center retrospective study

doi: 10.1186/s12885-024-12827-0

Figure Lengend Snippet: Pearson’s correlation analysis between serum TFPI2 and CA125 levels. There was a low correlation between serum TFPI2 and CA125 levels ( r = 0.203, p < 0.001). CA, cancer antigen; TFPI2, tissue factor pathway inhibitor 2

Article Snippet: Immunohistochemistry (IHC) was performed using the same concentration of mouse monoclonal anti-TFPI2 antibody (sc-48380, diluted 1:200, Santa Cruz Biotechnology, Inc., CA, USA) as the primary antibody to ensure consistency of our experimental approach [ ].

Techniques:

Journal: BMC Cancer

Article Title: Tissue factor pathway inhibitor 2 as a serum biomarker for endometrial cancer: a single-center retrospective study

doi: 10.1186/s12885-024-12827-0

Figure Lengend Snippet: Kaplan–Meier curve of preoperative serum TFPI2 levels and 5-year survival. Serum TFPI2 positivity is associated with poor prognosis in patients with endometrial cancer. P-value was calculated using the log-rank test (HR, 8.22; 95% CI, 2.49–27.1; p < 0.001). CI, confidence interval; HR, hazard ratio; TFPI2, tissue factor pathway inhibitor 2

Article Snippet: Immunohistochemistry (IHC) was performed using the same concentration of mouse monoclonal anti-TFPI2 antibody (sc-48380, diluted 1:200, Santa Cruz Biotechnology, Inc., CA, USA) as the primary antibody to ensure consistency of our experimental approach [ ].

Techniques:

Journal: BMC Cancer

Article Title: Tissue factor pathway inhibitor 2 as a serum biomarker for endometrial cancer: a single-center retrospective study

doi: 10.1186/s12885-024-12827-0

Figure Lengend Snippet: Multivariate analysis of prognostic factors for overall survival

Article Snippet: Immunohistochemistry (IHC) was performed using the same concentration of mouse monoclonal anti-TFPI2 antibody (sc-48380, diluted 1:200, Santa Cruz Biotechnology, Inc., CA, USA) as the primary antibody to ensure consistency of our experimental approach [ ].

Techniques:

Journal: BMC Cancer

Article Title: Tissue factor pathway inhibitor 2 as a serum biomarker for endometrial cancer: a single-center retrospective study

doi: 10.1186/s12885-024-12827-0

Figure Lengend Snippet: Microscopic immunohistochemistry images of TFPI2 expression in endometrial cancer tissues. Microscopic images show immunohistochemistry for TFPI2 in ( A ) clear cell carcinoma with intensity 0, ( B ) endometrioid carcinoma grade 3 with intensity 1+, ( C ) clear cell carcinoma with intensity 2+, and ( D ) serous carcinoma with intensity 3+. There were both IHC-positive and IHC-negative cases for all histological types. HE, hematoxylin eosin; IHC, immunohistochemistry; TFPI2, tissue factor pathway inhibitor 2

Article Snippet: Immunohistochemistry (IHC) was performed using the same concentration of mouse monoclonal anti-TFPI2 antibody (sc-48380, diluted 1:200, Santa Cruz Biotechnology, Inc., CA, USA) as the primary antibody to ensure consistency of our experimental approach [ ].

Techniques: Immunohistochemistry, Expressing

Journal: BMC Cancer

Article Title: Tissue factor pathway inhibitor 2 as a serum biomarker for endometrial cancer: a single-center retrospective study

doi: 10.1186/s12885-024-12827-0

Figure Lengend Snippet: Boxplots of IHC scores by each histological type. The Mann–Whitney U test showed no significant differences in IHC scores among histological types ( p = 0.427). CS, carcinosarcoma; EMG, endometrioid carcinoma grade; IHC, immunohistochemistry; TFPI2, tissue factor pathway inhibitor 2

Article Snippet: Immunohistochemistry (IHC) was performed using the same concentration of mouse monoclonal anti-TFPI2 antibody (sc-48380, diluted 1:200, Santa Cruz Biotechnology, Inc., CA, USA) as the primary antibody to ensure consistency of our experimental approach [ ].

Techniques: MANN-WHITNEY, Immunohistochemistry

Journal: Cancer Cell International

Article Title: Unveiling heterogeneity and prognostic markers in ductal breast cancer through single-cell RNA-seq

doi: 10.1186/s12935-024-03325-1

Figure Lengend Snippet: Univariate and multivariate Cox regression analysis for screening prognostic molecular markers and constructing a risk assessment model in breast cancer (BC). Note : ( A ) Lasso regression coefficient distribution; ( B ) Regression equation coefficients of 7 prognostic-related factors selected by Lasso regression; ( C ) Forest Figure visualizing univariate Cox analysis and multivariate Cox analysis of TCGA-BRCA cohort data; ( D – E ) Kaplan-Meier survival curves of CYP24A1 and TFPI2 genes; ( F ) Overall survival curve of TCGA-BRCA cohort patients grouped by Risk Score using Kaplan-Meier; ( G ) ROC curve depicting the predictive performance of the risk assessment model on TCGA-BRCA cohort; ( H ) ROC curve demonstrating the accuracy of the prognostic model constructed by gene expression of CYP24A1 and TFPI2 in predicting BC patients’ prognosis; ( I ) Kaplan-Meier survival curve of CYP24A1 gene; ( J ) Kaplan-Meier survival curve of TFPI2 gene

Article Snippet: The proteins were then transferred onto a PVDF membrane (1,620,177, BIO-RAD, USA) and blocked with 5% BSA at room temperature for 1 h. The primary antibodies, rabbit anti-CYP24A1 (PA5-21704, 1:1000, Thermo Fisher), TFPI2 (ab186747, 1:1000, abcam, UK), and GADPH (ab181602, 1:10000, abcam, UK) were applied and incubated overnight at 4 °C.

Techniques: Construct, Expressing

Journal: Cancer Cell International

Article Title: Unveiling heterogeneity and prognostic markers in ductal breast cancer through single-cell RNA-seq

doi: 10.1186/s12935-024-03325-1

Figure Lengend Snippet: The effects of overexpression of CYP24A1 or TFPI2 on the proliferation, migration, invasion, and apoptosis of BC cells. Note : ( A ) Expression of CYP24A1 and TFPI2 in BC tumor tissue and adjacent normal tissue was detected by RT-qPCR ( N = 35). ( B ) Expression of CYP24A1 and TFPI2 in BC tumor tissue and adjacent normal tissue was detected by IHC (400×, scale bar = 25 μm, N = 35). ( C ) mRNA expression levels of CYP24A1 and TFPI2 in regular human breast epithelial cell lines (MCF-10 A) and luminal BC cell lines (MCF-7 and T47D) were detected by RT-qPCR. ( D ) Protein expression of CYP24A1 and TFPI2 in regular human breast epithelial cell lines (MCF-10 A) and luminal BC cell lines (MCF-7 and T47D) was detected by Western blot. ( E – F ) RT-qPCR and Western blot detected the infection efficiency of overexpressed CYP24A1 and TFPI2 lentivirus in MCF-7 cells. ( G ) Cell proliferation of MCF-7 cells was assessed by CCK8 assay. ( H ) Cell apoptosis of MCF-7 cells was detected by flow cytometry. ( I ) Cell migration and invasion of MCF-7 cells were examined by Transwell assay. The data in Figure are all measurement data, represented by sample mean ± standard deviation; paired t-test is used for Comparison between cancer tissue and adjacent normal tissue, independent sample t-test is used for Comparison between two groups, and one-way analysis of variance is used for Comparison between multiple groups Comparison of inter-group data at different time points, cell viability was analyzed using a two-way analysis of variance. * represents the Comparison between two groups, with P < 0.05

Article Snippet: The proteins were then transferred onto a PVDF membrane (1,620,177, BIO-RAD, USA) and blocked with 5% BSA at room temperature for 1 h. The primary antibodies, rabbit anti-CYP24A1 (PA5-21704, 1:1000, Thermo Fisher), TFPI2 (ab186747, 1:1000, abcam, UK), and GADPH (ab181602, 1:10000, abcam, UK) were applied and incubated overnight at 4 °C.

Techniques: Over Expression, Migration, Expressing, Quantitative RT-PCR, Western Blot, Infection, CCK-8 Assay, Flow Cytometry, Transwell Assay, Standard Deviation, Comparison

Journal: Cancer Cell International

Article Title: Unveiling heterogeneity and prognostic markers in ductal breast cancer through single-cell RNA-seq

doi: 10.1186/s12935-024-03325-1

Figure Lengend Snippet: The effect of CYP24A1 or TFPI2 overexpression on tumorigenic and metastatic abilities in BC cells. Note : ( A ) Growth curves of tumor masses in nude mice in each group; ( B ) Representative Figures of tumor masses in nude mice in each group; ( C ) Comparison of tumor mass weights in nude mice in each group; ( D ) Observation of lung metastasis of tumor masses in nude mice in each group through H&E staining (10×, scale bar = 1 cm.); ( E ) Counting of lung metastatic nodules in nude mice in each group. The data in Figure are all metric data, represented by sample mean ± standard deviation. Single-factor analysis of variance is used for Comparison among multiple groups. Comparison of data between different groups at different time points is conducted using repeated measures analysis of variance. * represents the Comparison between two groups, with P < 0.05. N = 6

Article Snippet: The proteins were then transferred onto a PVDF membrane (1,620,177, BIO-RAD, USA) and blocked with 5% BSA at room temperature for 1 h. The primary antibodies, rabbit anti-CYP24A1 (PA5-21704, 1:1000, Thermo Fisher), TFPI2 (ab186747, 1:1000, abcam, UK), and GADPH (ab181602, 1:10000, abcam, UK) were applied and incubated overnight at 4 °C.

Techniques: Over Expression, Comparison, Staining, Standard Deviation

Journal: Cancer Cell International

Article Title: Unveiling heterogeneity and prognostic markers in ductal breast cancer through single-cell RNA-seq

doi: 10.1186/s12935-024-03325-1

Figure Lengend Snippet: The molecular regulation of CYP24A1 and TFPI2 in the growth and metastasis of breast cancer

Article Snippet: The proteins were then transferred onto a PVDF membrane (1,620,177, BIO-RAD, USA) and blocked with 5% BSA at room temperature for 1 h. The primary antibodies, rabbit anti-CYP24A1 (PA5-21704, 1:1000, Thermo Fisher), TFPI2 (ab186747, 1:1000, abcam, UK), and GADPH (ab181602, 1:10000, abcam, UK) were applied and incubated overnight at 4 °C.

Techniques:

Journal: Nature Communications

Article Title: Qualitative differences in disease-associated MEK mutants reveal molecular signatures and aberrant signaling-crosstalk in cancer

doi: 10.1038/s41467-022-31690-w

Figure Lengend Snippet: a Transcriptome analyses of HEK293 cells stably expressing HA-MEK1 WT, F53S, or K57N. Genes differentially expressed between WT and F53S (left), WT and K57N (middle), and F53S and K57 (right) are displayed as volcano plots ( x -axis: fold-change [log2], y -axis: p -value [-log10]). Red area, log 2 FC > 1 and p < 0.05 ( n = 3); blue area, log 2 FC < −1 and p < 0.05 ( n = 3). b Venn diagrams illustrating the number of genes significantly upregulated (>2-fold; left) and downregulated (<0.5-fold; right) in F53S and K57N cells vs. WT cells. c Principal component analysis of the gene expression profiles of WT, F53S, and K57N cells. d KEGG pathway enrichment analyses of genes upregulated in F53S and K57N cells vs. WT cells. e Gene sets enriched analysis. The indicated gene signatures were significantly enriched in K57N cells compared with F53S cells. f Five distinct patterns of gene expression relative to the degree of aberrant ERK activity in F53S and K57N cells. (Upper) Schematic representations of the gene expression patterns. (Lower) qRT-PCR analysis of the expression level of a representative gene in each group. Data are mean ± SEM from three independent experiments. P -values were determined using one-way ANOVA followed by Tukey’s multiple comparisons test. ns, not significant. g Immunoblot analysis of representative secreted proteins of the sigmoidal-B group (TFPI2 and GDF15) and the bell-shaped group (COL14A1) in cell culture supernatants from the indicated cells. Source data are provided as a Source Data file.

Article Snippet: The following primary antibodies were used: Monoclonal anti-HA F-7 (Santa Cruz Biotechnology, sc-7392), anti-GST B-14 (sc-138), anti–Myc 9E10 (sc-40), anti-TFPI2 B-7 (sc-48380), anti-MEK1 H-8 (sc-6250), anti-HA 16B12 (Covance, MMS-101R), anti-HA 3F10 (Roche, 11867423001), anti-Flag M2 (Sigma–Aldrich, F1804), anti-phospho-Raf-1 56A6 (Cell Signaling Technology, 9427), anti-phospho-MEK1/2 41G9 (9154), anti-phospho-AKT1(T308) 244F9 (4056), anti-phospho-AKT1(S473) D9E (4060), anti-phospho-S6 D57.2.2E (4858), anti-S6 5G10 (2217), anti-MEK2 13E3 (9147), anti-GDF15 D2A3 (8479), anti-PHLDA1 EPR6674 (Abcam, ab133654), anti-PHLDA2 (ab58379), Anti-β-Actin 6D1 (FUJIFILM Wako, 010-27841); polyclonal anti-ERK (Santa Cruz Biotechnology, sc-94), anti-C-Raf-1 (sc-227), anti-B-Raf (sc-166), anti-phospho-ERK1/2 (Cell Signaling Technology, 9101), anti-AKT (9272), anti-cleaved caspase3 (9661), anti-PARP (9542), anti-COL14A1 (Abcam, ab101464), anti-TM4SF1 (ab113504), anti-PHLDA2 (Proteintech, 14661-1-AP), anti-TM4SF19 (Sigma-Aldrich, SAB1102826), anti-EMP1 (SAB1302714).

Techniques: Stable Transfection, Expressing, Activity Assay, Quantitative RT-PCR, Western Blot, Cell Culture