ABSTRACT
Objective@#To detect the expressions of epidermal growth factor-like domain 7 (EGFL7) and P185 protein in breast cancer tissues and serum, and to analyze the correlation between the expression levels of EGFL7 and P185 in tissues and clinicopathological parameters of breast cancer patients.@*Methods@#Sixty patients with breast cancer in Hunan Cancer Hospital from March 2016 to March 2018 were collected as observation group, and 60 patients with breast benign lesions in the hospital during the same period were selec-ted as control group. The expressions of EGFL7 and P185 protein in tissues of patients in the two groups were detected by immunohistochemical two-step method, and the levels of EGFL7 and P185 protein in serum of patients in the two groups were detected by enzyme-linked immunosorbent assay (ELISA). The relationships between the expressions of EGFL7 and P185 protein and clinicopathological parameters of breast cancer patients were analyzed.@*Results@#The positive rates of EGFL7 in tissues in the observation group and the control group were 65.00% (39/60) and 28.33% (17/60), and there was a significant difference between the two groups (χ2=16.205, P<0.001). The positive rates of P185 in tissues in the two groups were 43.33% (26/60) and 15.00% (9/60), and there was a significant difference between the two groups (χ2=11.657, P=0.001). The serum levels of EGFL7 protein in the observation group and the control group were (3.39±0.38) μg/ml and (2.75±0.31) μg/ml respectively, with a significant difference (t=10.109, P<0.001). The serum levels of P185 protein in the two groups were (7.12±0.75) μg/ml and (6.08±0.62) μg/ml respectively, with a significant difference (t=8.279, P<0.001). The positive expression of EGFL7 protein was closely related to tumor size (χ2=6.128, P=0.013), TNM stage (χ2=7.781, P=0.005) and metastasis (χ2=5.444, P=0.020). The positive expression of P185 protein was closely related to tumor size (χ2=8.910, P=0.003) and TNM stage (χ2=8.024, P=0.005).@*Conclusion@#The levels of EGFL7 and P185 protein are high in breast cancer tissues and serum, and their positive expressions are related to tumor size and TNM stage. EGFL7 and P185 proteins play important roles in the progression of breast cancer.
ABSTRACT
Objective To detect the expressions of epidermal growth factor-like domain 7 (EGFL7) and P185 protein in breast cancer tissues and serum,and to analyze the correlation between the expression levels of EGFL7 and P185 in tissues and clinicopathological parameters of breast cancer patients. Methods Sixty patients with breast cancer in Hunan Cancer Hospital from March 2016 to March 2018 were collected as observation group,and 60 patients with breast benign lesions in the hospital during the same period were selec-ted as control group. The expressions of EGFL7 and P185 protein in tissues of patients in the two groups were detected by immunohistochemical two-step method,and the levels of EGFL7 and P185 protein in serum of patients in the two groups were detected by enzyme-linked immunosorbent assay (ELISA). The relationships between the expressions of EGFL7 and P185 protein and clinicopathological parameters of breast cancer patients were analyzed. Results The positive rates of EGFL7 in tissues in the observation group and the control group were 65. 00％ (39 / 60)and 28. 33％ (17 / 60),and there was a significant difference between the two groups (χ2 = 16. 205,P < 0. 001). The positive rates of P185 in tissues in the two groups were 43. 33％ (26 / 60)and 15. 00％ (9 / 60),and there was a significant difference between the two groups (χ2 = 11. 657,P = 0. 001). The serum levels of EGFL7 protein in the observation group and the control group were (3. 39 ± 0. 38)μg/ ml and (2. 75 ± 0. 31)μg/ ml respectively,with a significant difference (t = 10. 109,P < 0. 001). The serum levels of P185 protein in the two groups were (7. 12 ± 0. 75)μg/ ml and (6. 08 ± 0. 62)μg/ ml respectively, with a significant difference (t = 8. 279,P < 0. 001). The positive expression of EGFL7 protein was closely related to tumor size (χ2 = 6. 128,P = 0. 013),TNM stage (χ2 = 7. 781,P = 0. 005)and metastasis (χ2 =5. 444,P = 0. 020). The positive expression of P185 protein was closely related to tumor size (χ2 = 8. 910, P = 0. 003)and TNM stage (χ2 = 8. 024,P = 0. 005). Conclusion The levels of EGFL7 and P185 protein are high in breast cancer tissues and serum,and their positive expressions are related to tumor size and TNM stage. EGFL7 and P185 proteins play important roles in the progression of breast cancer.
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Objective: To explore the effects of epidermal growth factor-like domain 6 (EGFL 6) gene silencing by specific small interference RNA (siRNA) on the proliferation and invasion of human melanoma cells. Methods: The double-strand siRNA targeting EGFL 6 gene (named as EGFL6-siRNA) was designed and synthesized. The EGFL6-siRNA was transfected into human melanoma A375 cells by liposome transfection method. The mRNA and protein expression levels of EGFL6 in A375 cells were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The proliferation and invasion abilities of A375 cells were analyzed by cell counting kit-8 (CCK-8) and Transwell assay, respectively. Results: After EGFL6-siRNA was successfully transfected into human melanoma A375 cells for 48 h, the EGFL6 mRNA and protein expression levels in A375 cells were significantly decreased (both P < 0.01). The EGFL6-siRNA could significantly inhibit the proliferation of A375 cells at different time points (24-72 h) after transfection (all P < 0.01), and significantly inhibit the invasion of A375 cells (P < 0.01). Conclusion: The specific siRNA can effectively silence EGFL 6 gene expression in human melanoma A375 cells, and suppress the proliferation and invasion of melanoma cells.
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ObjectiveTo test the expression of epidermal growth factor-like domain 7 (EGFL7),microvessel density (MVD) and foeal adhesion kinase pY397 (FAKpY397) in human glioma tissues,and to evaluate their relationship.MethodsThe expression of EGFL7 and FAKpY397 in 56 cases of human glioma and 8 cases of normal brain tissues were detected by immunohistochemistry test,and MVD was detected by CD34 staining.ResultsThere was a significant difference of the positive rates of EGFL7 between normal brain tissue (0) and gliomas (75％),χ2 =17.45,P ＜0.01.With the increased pathological grade,the expression level of EGFL7 increased (χ2 =26.24,P ＜ 0.01 ).There was a significant difference of the positive rates of FAKpY397 between normal brain tissue ( 12.5％ ) and gliomas (73.2％),χ2 =6.23,P ＜ 0.05.With the increased pathological grade,the expression level of FAKpY397 increased (χ2 =6.71,P ＜ 0.01 ). MVD on normal brain was( 15 ± 4 )/HP,on Ⅰ - Ⅱ grade and Ⅲ -Ⅳ grade gliomas was ( 27 ± 3 )/HPand ( 60 ± 4 )/HP respectively,there was a significant difference on MVD between normal brain tissue and gliomas (P ＜ 0.01 ).Higher level of MVD was found in gliomas with higher grade ( P ＜ 0.01 ).There was a positive correlation between EGFL7 and FAKpY397 expressions in gliomas (r =0.314,P ＜0.01 ).There was a significant difference on MVD between positive and negative expression of EGFL7 ( t =26.55,P ＜ 0.01 ). MVD was (56 ± 4 )/HP and (25 ± 3 )/HP respectively.ConclusionThe expression of EGFL7 of human gliomas has a favorable positive correlation with the degree of malignancy,MVD and FAKpY397.It is indicated that EGFL7not only palys an important regulative role in glioma neovascularization,but also it may participate directly in glioma occurrence and invasion.
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The binding function of EGF1 domain peptide with tissue factor(TF)and its ability of triggering coagulation were explored.The TF expression model in vitro was established by lipopolysaccharide induction.The affinity of EGFP-EGF1 and TF expressing cells was analyzed by fluorescence microscopy and flow cytometry(FCM).The affinity of EGFP-EGF1 and rat soluble TF was quantitated by surface plasmon resonance(SPR).The ability of EGFP-EGF1 in triggering coagulation was tested by prothrombin time assay.The FCM results showed recombinant factor Ⅶ(rFⅦ)could definitely depress the integration of EGFP-EGF1 with recombinant TF(rTF)(68.65%±3.86% vs 57.98%±4.71%,P<0.01).The SPR results indicated the association constant ka of EGFP-EGF1 proteins was higher than rFⅦ(8.29±1.39 vs 3.75±0.32,P<0.01).However,the EGFP-EGF1 protein lost the activity of triggering coagulation as compared with blood plasma of normal SD rats(56.8±3.2 s vs 17.8±3.4 s,P<0.01).It was concluded that the rat EGF1 peptide could specifically bind to TF without the ability of triggering coagulation.EGF1 peptide may be a good target head for delivering drugs to TF in anticoagulation therapy.