The Functional Proteomics Analysis Of VEGF-treated Human Epithelial Ovarian Cancer Cells HO-8910

Background and Objective:In gynecological reproductive tract tumors,the incidence of ovarian cancer is lower than cervical cancer and metrocarcinoma,but it is the highest level of mortality.VEGF(vascular endothelial growth factor) as the most important angiogenic factor,plays a key role in variety tumor malignant progression and transform.A number of studies have confirmed that VEGF is closely related to the invasion and metastasis of ovarian cancer and some of them show that tumor cells secrete high levels of VEGF factor in paracrine or autocrine manner to the tumor cells themselves,which impact the tumor cell's proliferation,invasion capacity or its specific mechanisms are not clear. In order to investigate the relationship between VEGF associated proteins and ovarian cancer,our study is to compare the differentially proteins between the ovarian cancer HO-8910 cells and the VEGF stimulated ovarian cancer HO-8910 cells,then screening the VEGF related proteins.Methods:(1) HO-8910 as the research object,routinely cultured ovarian cancer cell line HO-8910,VEGF stimulated HO-8910 cells for 24 h as the experimental group,the control group HO-8910 cells without VEGF stimulation.(2) Two-dimensional electrophoresis(2-DE) separate the two groups proteins,PD Quest image analysis software identify differentially expressed proteins,matrix-assisted laster desorption/ ionization time-of-flight mass spectrometry(MALDI-TOF-MS) identification of differentially expressed proteins.(3) The real time PCR verified mRNA different expression level of some differentially expressed proteins.(4) Western blot and immunocytochemistry were used to verify some of the differentially expressed proteins.(5) Transmission electron microscopy observed ultrastructure changes of two groups.Results:(1) Established the 2-DE map of the experimental group and control group cells,image analysis identified 30 differentially expressed protein spots,mass spectrometry identified 17 non-redundant proteins.(2) Real time PCR verified the differentially expression proteins Tim,PPIase A,Lamin-A/C and Ezrin.(3) Western blot and Immunocytochemistry verified the differentially expression of Ezrin.(4) Transmission electron microscope observed that the malignant degree of HO-8910/VEGF is increased.Conclusions:(1) Established the 2-DE maps of the experimental group and control group cells,Image analysis identified 30 differentially expressed protein spots,mass spectrometry identified 17 non-redundant proteins,their functions involved in tumor cell invasion,transfer, apoptosis and proliferation.(2) VEGF down-regualted the expression of Ezrin,PPIase A and Lamin-A/C in VEGF stimulated HO-8910 cells and up-reguated the Tim expression.(3) After VEGF inducation,the malignant degree of HO-8910 is increased.(4) These results suggest that, VEGF may be influence the invasion of ovarian cancer through Ezrin, PPIase A,Lamin-A/C and Tim.It may have a potential clinical value.