egfl7 epidermal growth factor-like protein  (Angiocrine)

Journal: Scientific Reports

Article Title: Circulating EGFL7 distinguishes between IUGR and PE: an observational case–control study

doi: 10.1038/s41598-021-97482-2

Figure Lengend Snippet: EGFL7 levels in non-pregnant women, healthy pregnant women, and pregnant women affected by IUGR, Early PE and Late PE. ( A ) Mean (± SE) levels of Epidermal Growth Factor-like Domain 7 (EGFL7) in plasma of non-pregnant women (NP), healthy pregnant women (Controls), IUGR-, Early PE- (e-PE) and Late PE- (l-PE) affected pregnant women throughout pregnancy as measured by ELISA. Data were compared using the two-sample t -test between two samples ( p < 0.001 for e-PE vs. CTRL and IUGR at GA 22–25; p = 0.05 for e-PE vs. CTRL and IUGR at GA 26–30; p = 0.045 for e-PE vs. IUGR at GA 31–34; p = 0.01 for l-PE vs. CTRL) or ANOVA test among the three studied groups ( p < 0.001 at GA 22–25 and p = 0.036 at GA 35–40). Statistically significant difference within gestational interval is reported. SigmaPlot 12.0 and GraphPad Prism 7 were used for statistical analysis. ( B ) Real-time PCR analysis for EGFL7 gene expression levels in placental samples obtained from Controls, IUGR- and l-PE- affected pregnant women at GA 35–40. Data were represented as mean (± SE). ANOVA test with post-hoc Bonferroni correction was used for statistical analysis (SigmaPlot 12.0). Differences were considered significant at p < 0.05.

Article Snippet: Human EGFL7 kits were purchased from Cusabio Biotech (College Park, Maryland, USA).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Gene Expression

Journal: Cancers

Article Title: The Multifaceted Roles of EGFL7 in Cancer and Drug Resistance

doi: 10.3390/cancers13051014

Figure Lengend Snippet: ( a ) Model structure of the EGFL7 protein. EGFL7 contains two EGF-like repeats: an arg-gly-asp integrin-binding motif (RGD) and an Emilin-like region. ( b ) The regulatory network of EGLF7. EGFL7 binds via its RGD domain to ITGAV:ITGB3 integrin and causes among others FAK autophosphorylation. EGFL7 competes for binding to the integrin with the ECM molecules fibronectin, MMP2, and collagen IV. The Emilin-like region of EGFL7 interacts with Notch receptor 1–4 and Notch ligands DLL4 and Jag1 and suppresses Notch signaling, resulting in impaired NIC translocation into the nucleus and reduced Hey1/2 and Hes1 transcription. EGFL7 binding to EGFR results in the activation of the signaling pathways extracellular signal-regulated kinase (ERK) and AKT, among others. EGFL7 competes for binding to EGFR with EGF. Abbreviations: EGF, epidermal growth factor; EGFL7, epidermal growth factor-like protein 7; EGFR, epidermal growth factor receptor; ERK, extracellular signal-regulated kinase; FAK, focal adhesion kinase); MMP2, matrix metalloproteinase 2; DLL4, delta like-protein 4; Jag1, Jagged1; NIC, Notch intracellular domain; ECM, extracellular matrix.

Article Snippet: ECs release so-called angiocrine factors [ ], which include the angiogenic factor vascular endothelial growth factor-A (VEGF-A), Jagged1 (Jag1), endothelin, enzymes such as tissue-type plasminogen activator [ ], and epidermal growth factor-like protein-7 (EGFL7) [ ].

Techniques: Binding Assay, Translocation Assay, Activation Assay, Protein-Protein interactions

Journal: Cancers

Article Title: The Multifaceted Roles of EGFL7 in Cancer and Drug Resistance

doi: 10.3390/cancers13051014

Figure Lengend Snippet: EGFL7 alters tumor growth and metastasis by suppressing the production of immune cells and their recruitment into the growing tumors, vessel permeability, ECM stiffness—all of which contribute to drug resistance. T cell adhesion and rolling and transmigration through the EC are required for T cells to cross the vascular barrier. EGFL7 allows the tumor to escape from the anti-tumor immune response by preventing terminal T cell differentiation in the thymus and inhibiting T cell recruitment via suppression of the adhesion molecules ICAM and VCAM on ECs. EGFL7 prevents adhesion molecule transcription after tumor necrosis factor alpha (TNFa) stimulation by blocking Nuclear factor kappa B (NFkB) signaling. EGFL7 controls ECM stiffness by interacting with LOXL2 so as to mitigate covalent crosslinking of collagen or elastin. ITGB1 integrin on ECs and ITGB3 integrin on cancer cells induce the expression of transcription factor KLF2 which enhances EGFL7 expression, resulting in enhanced cell proliferation. Myelosuppressive drugs such as bortezomib were shown to enhance KLF2-mediated upregulation of ITGB3 and EGFL7. Abbreviations: KLF2, Krüppel-like factor 2; ETP, early thymic progenitor; LOXL2, lysyl oxidase-like 2.

Article Snippet: ECs release so-called angiocrine factors [ ], which include the angiogenic factor vascular endothelial growth factor-A (VEGF-A), Jagged1 (Jag1), endothelin, enzymes such as tissue-type plasminogen activator [ ], and epidermal growth factor-like protein-7 (EGFL7) [ ].

Techniques: Permeability, Transmigration Assay, Cell Differentiation, Blocking Assay, Expressing

Journal: Frontiers in Oncology

Article Title: Novel Expression of EGFL7 in Osteosarcoma and Sensitivity to Cisplatin

doi: 10.3389/fonc.2020.00074

Figure Lengend Snippet: Compared with CS, EGFL7 is highly expressed in OS. (A) Typical pathological features of OS and CS revealed by HE staining, tumor-like osteogenesis can be seen in OS and cartilage-like matrix in CS. (B,C) Immunohistochemical of EGFL7 protein expression in OS tissue ( n = 2) and CS controls ( n = 2), comparison of IHC results from different magnification (100 and 200 ×) and its significance. High expression of EGFL7 (++) was found in OS tissues but negative in CS, the arrow refers to the positive area. The semi-quantitative statistical analysis based on IHC results shows that there is a significant difference in the expression of EGFL7 protein between OS and CS. (D,E) Western blot results showed that EGFL7 protein was overexpressed in OS tissue compared with CS, Student's t -test shows that the difference between OS and CS has obvious statistical significance. The abundance of EGFL7 protein in OS tissue was similar to that of GAPDH protein. (F) RT-PCR results showed that the transcription level of EGFL7 mRNA in OS tissues was significantly higher than that in CS tissues, and the difference was statistically significant. (G) Immunohistochemical results of chondrosarcoma and osteosarcoma showed that EGFL7 was highly expressed in osteosarcoma. (H) The immunohistochemical results of chondrosarcoma and osteosarcoma showed that CD34 was highly expressed in osteosarcoma, indicating that there were more abundant blood vessels in osteosarcoma. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The level of EGFL7 protein in cell culture medium was determined using an ELISA kit (CUSABIO, Wuhan, China).

Techniques: Staining, Immunohistochemical staining, Expressing, Comparison, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Frontiers in Oncology

Article Title: Novel Expression of EGFL7 in Osteosarcoma and Sensitivity to Cisplatin

doi: 10.3389/fonc.2020.00074

Figure Lengend Snippet: The primers used for Q-PCR

Article Snippet: The level of EGFL7 protein in cell culture medium was determined using an ELISA kit (CUSABIO, Wuhan, China).

Techniques:

Journal: Frontiers in Oncology

Article Title: Novel Expression of EGFL7 in Osteosarcoma and Sensitivity to Cisplatin

doi: 10.3389/fonc.2020.00074

Figure Lengend Snippet: Upregulation of EGFL7 expression in OS cell line compared with ECs and CS cell line. (A,B) Western blot results showed that EGFL7 protein was overexpressed in all OS cell lines and HUVECs compared to SW1353. Statistical analysis showed that the expression of EGFL7 in OS cell lines was significantly different from that in SW1353. Similarly, the expression of EGFL7 protein in HUVEC was significantly different from that in SW1353. (C) RT-PCR results showed that the transcription level of EGFL7 mRNA in OS cell lines was significantly higher than HUVEC, there are also differences between HUVEC and SW1353, and the difference was all statistically significant. PCR and western blot experiments were all performed in triplicate. (D) The ELISA results of EGFL7 in cell culture medium also confirmed that there was indeed a high expression of EGFL7 protein in osteosarcoma cells. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The level of EGFL7 protein in cell culture medium was determined using an ELISA kit (CUSABIO, Wuhan, China).

Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Frontiers in Oncology

Article Title: Novel Expression of EGFL7 in Osteosarcoma and Sensitivity to Cisplatin

doi: 10.3389/fonc.2020.00074

Figure Lengend Snippet: Immunofluorescence technique was used to verify the expression of EGFL7 from histological and cytological aspects, respectively. (A) As seen in these representative images, the EGFL7 was labeled by special anti-EGFL7 antibody, and the cell nuclei were stained by DAPI. The merged image shows that green fluorescent signals for EGFL7 protein were expressed in both tumor tissues and vascular endothelial cells, and we have observed this expression from multiple samples and different optical multiples, respectively. White arrows indicate positive areas. (B) From the immunofluorescence results of OS cells, the EGFL7 protein was labeled by red fluorescent, the cell nuclei were stained by DAPI. The expression of EGFL7 protein in OS cells is located in the cytoplasm.

Article Snippet: The level of EGFL7 protein in cell culture medium was determined using an ELISA kit (CUSABIO, Wuhan, China).

Techniques: Immunofluorescence, Expressing, Labeling, Staining

Journal: Frontiers in Oncology

Article Title: Novel Expression of EGFL7 in Osteosarcoma and Sensitivity to Cisplatin

doi: 10.3389/fonc.2020.00074

Figure Lengend Snippet: The expression of EGFL7 deregulated in OS cells after cisplatin intervention. (A) Intervention of four osteosarcoma cell lines with different gradient concentration of cisplatin. Under light microscope, the number of cells changed significantly with different concentrations of cisplatin. The toxicity of cisplatin was detected by CCK-8 kit and the corresponding IC50 of various cells was calculated. (B) The results showed that cisplatin could significantly affect the proliferation rate of tumor cell lines. (C,D) Western blot was assessed to compare EGFL7 protein in OS cells before and after Cisplatin intervention, the results showed that the expression of EGFL7 protein deregulated after Cisplatin intervention. Paired T test showed that EGFL7 protein expression in OS cells was significantly deregulated after cisplatin intervention and the difference was statistically significant. (E) RT-PCR results showed that the transcription level of EGFL7 mRNA in OS cell lines was significantly deregulated after cisplatin intervention and the difference was statistically significant. PCR and western blot experiments were all performed in triplicate. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The level of EGFL7 protein in cell culture medium was determined using an ELISA kit (CUSABIO, Wuhan, China).

Techniques: Expressing, Concentration Assay, Light Microscopy, CCK-8 Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Frontiers in Oncology

Article Title: Novel Expression of EGFL7 in Osteosarcoma and Sensitivity to Cisplatin

doi: 10.3389/fonc.2020.00074

Figure Lengend Snippet: Chemotherapy deregulated expression of EGFL7 in osteosarcoma. (A) Samples obtained by biopsy before chemotherapy and by tumor resection after chemotherapy in the same patient. Pre-chemotherapy tissue specimens showed fish-like appearance, while post-chemotherapy tumor tissue showed obvious bone repair changes. (B,C) Immunohistochemical staining of the same patient's tumors before and after chemotherapy showed that there was almost no expression of EGFL7 in the tumors after chemotherapy (black arrows represent positive areas). The semi-quantitative statistical analysis based on IHC results shows that there is a significant difference in the expression of EGFL7 between pre-chemotherapy and post-chemotherapy. (D,E) Western blot results showed that EGFL7 protein was almost no expression in post-chemotherapy tissue compared with pre-chemotherapy tissue, Student's t -test shows that the difference has obvious statistical significance. (F) RT-PCR results showed that the transcription level of EGFL7 mRNA in post-chemotherapy tissue was significantly deregulated than that in pre-chemotherapy tissue, and the difference was statistically significant. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The level of EGFL7 protein in cell culture medium was determined using an ELISA kit (CUSABIO, Wuhan, China).

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