Effects Of MIF On The Cell Proliferation And Invasion Abilities As Well As On VEGF Expression In MCF-7 Cells

Objective: One of the aims is to investigate the influence of down-regulation of macrophage migration inhibitory factor(MIF)expression on the proliferation and invasion ability of MCF-7 cells and another is to investigate the influence of down-regulation of MIF expression on the proliferation ability and expression of VEGF in xenograft tumors.Methods: 1.Proliferation abilities were detected by CCK-8 assay and flow cytometer detections were used to assess the cell cycle with knocking down MIF in MCF-7 cells.Besides,the expression of Cyclin D1 was measured by the means of Western blot.2.The migratory ability of siMIF MCF-7 cells was tested with scar assay and Transwell method.The invasion ability of siMIF MCF-7 cells was tested with Matrigel invasion assay.In addition,the expression of MMP14 protein in breast cancer cells was evaluated by Western blot.3.After MIF knocked down in MCF-7 cells,the protein levels of p-ERK and p-P38 were measured with Western blot.4.After the xenograft tumors model was established,the volume was measured and growth curves was drawn.5.Western blot was used to detect the expression levels of p-ERK and p-P38 proteins of BALB/c mice xenograft tumor tissues.6.Expression levels of CD31 and D2-40 in mice xenograft tumor tissues were detected by immunohistochemistry with MIF knocking down,and evaluate the micro vessel density together with the density of micro lymphatic vessel.Results: 1.After siRNA effectively blocked the MIF gene transcription and translation in MCF-7 breast cancer cells,the cell vitality was restrained and the cell cycle stagnated in G1-S period,besides the expression level of Cyclin D1 protein is lower than in the MCF-7 cell.2.The cell number passed through the basement membrane(with or without matrigel)of siMIF MCF-7 cells is lower than controls,as well as the level of MMP14.3.After MIF gene knocked down,the expression level of p-ERK and p-P38 protein was decreased,consequently.4.The tumors of xenograft tumor models established with siMIF MCF-7 cells were smaller and lighter than controls.5.The expression level of p-ERK and p-P38 protein was decreased xenograft tumor models established with siMIF MCF-7 cells.6.Low expression of MIF cells of BALB/c mice transplanted tumor model of VEGF-A and VEGF-C transcription and translation levels were lower than the control groups.The count of micro vessel and micro lymphatic vessel density were less than the control groups obviously.Conclusions: 1.Expression level of MIF in MCF-7 cells in breast cancer were positively correlated with cell proliferation,migration as well as invasion ability,p-P38 and p-ERK proteins of MAPK signaling pathway play a role in this process.2.Expression level of MIF of BALB/c mice xenograft tumors and transplanted tumor proliferation together with number of micro vessel and micro lymphatic vessel were positively correlated.