The Study Of Molecular Mechanism That Interleukin-8 Receptor Antagonist Inhibiting Prostate Cancer Invasion

ObjectivesIn this study,we investigated the molecular mechanism of IL-8 / CXCR2 receptor antagonist SB225002 on the invasion ability of prostate cancer cells LNCAP,DU-145 and PC-3 by in vitro cell experiments and animal experiments.Methods1.Prostate cancer tissue chip,the expression of IL-8 and BSP in different stages,with/without distant and lymphatic metastasis were detected by immunohistochemistry and the correlation between them was analyzed;2.To culture androgen-dependent prostate cancer cells LNCAP and androgen-independent cancer cells DU-145 and PC-3,using MTT assay SB22500 growth inhibition of three cell lines;3.Transwell and migration experiment used to detection the effect of SB225002 on three cell invasion and migration;4.Three cell lines were divided into SB225002 treatment group and control group.The treatment group was treated with SB225002(10μM),and the control group was treated with the same amount of DMSO.After cultured for 72 h,the expression of invasion related proteins MMP-2,MMP-9,BSP and αVβ3 changes were detected by Western blot;5.Three cells were treated with PI3 K signaling pathway inhibitor,Western bolt used to verify whether the PI3 K signaling pathway can regulate the expression of those invasion-related proteins.Finally,the expression and phosphorylation of PI3 K signaling pathway proteins change after SB225002 treated were detected by Western blot;6.Animal experiment: Prostate cancer xenograft models were constructed by subcutaneous implantation of prostate cancer cells in mice.The mice were divided into three group: negative control group,a subcutaneous implantation group,and a planting + administration group.The mice in the implantation + administration group were injected with SB225002 by intraperitoneal injection.The subcutaneous tumor volume changes in the mice during the administration were measured and administered for a period of time,The mice were sacrificed to remove the tumor.Immunohistochemistry was used to detect the expression of four invasion-related proteins in the xenograft tumor tissues.At the same time,the ocular arterial blood was taken from the mice and the serum concentrations of BSP in the three groups were detected by ELISA.Results1.The expression of IL-8 and BSP were high expression in prostate cancer tissue with distant metastasis,and low-moderate expression in the tissue without metastasis,and the expression level of IL-8 and BSP in prostate cancer tissues has moderately related.2.IL-8 receptor antagonist can significantly inhibit the proliferation of androgen-independent prostate cancer cells.SB225002 can significantly reduce the invasion ability of prostate cancer cells,and reduce the expression of MMP-2 and BSP in prostate cancer cells.3.PI3 K signaling pathway inhibitor can reduce the expression of MMP-2,BSP and MMP-9;SB225002 can reduce the phosphorylation level of AKT and mTOR,indicating that SB225002 may inhibit the invasion through suppression PI3 K signaling pathway.4.SBB225002 can inhibit the growth of prostate cancer xenografts in vivo and reduce the expression of BSP and MMP-2 in tumor tissues.While inhibiting the secretion of BSP by tumor cells.ConclusionIL-8 / CXCR2 receptor antagonist can inhibit the proliferation of prostate cancer cells and reduce its invasive ability.It can inhibit the invasion of cancer cells by inhibiting the phosphorylation of AKT / mTOR and decreasing the expression levels of invasion related proteins MMP-2 and BSP.