The inhibition effects and mechanism of recombinant snake venom metalloproteinase inhibitor on tumor angiogenesis
10.3969/j.issn.1001-1978.2017.03.020
- VernacularTitle:蛇毒金属蛋白酶抑制剂重组蛋白抗肿瘤血管生成作用及其机制
- Author:
Mingkai JI
;
Lihong CHEN
;
Bo CHENG
;
Yi SHI
;
Xu LIN
;
Jianyin LIN
- Keywords:
snake venom;
matrix metalloproteinase;
recombinant protein;
expression and purification;
neo-vascularization;
signal transduction
- From:
Chinese Pharmacological Bulletin
2017;33(3):394-400
- CountryChina
- Language:Chinese
-
Abstract:
Aim Toinvestigatetheeffectofrecombi-nant snake venom metalloproteinase inhibitor (rSVM-PI ) on neovascularization and its molecular mecha-nism.Methods Chickenchorioallantoicmembrane (CAM)assay was used to examine the antiangiogenic effect of rSVMPI.Alamar blue analysis was used to de-tect cell proliferation.Annexin V-FITC double labeling flow cytometry was used to assay cell apoptosis. Scratch marker was used to assay cell migration.Boy-den chamber analysis method was used to detect cells chemotaxis in vitro.Tube like structure(TLS)of HU-VECs was used to detect the ability of neovasculariza-tion in vitro.Real-time PCR and Western blot were used to assay the expressions of KDR and FGFR-1 inHUVECs. Results Thevasculardensityindex (VDI)of CAM was drastically decreased after rSVMPI treatment, chemotaxis of HUVECs in response of VEGF was inhibited in the presence of rSVMPI,TLS of HUVECs was less than control group.The expres-sions of KDR and FGFR-1 were down-regulated by re-al-timePCRandWesternblotassay.Conclusion rS-VMPI may inhibit neovascularization by blocking the VEGF-KDR or bFGF-FGFR signal transduction path-way.
