Sema4C Regulates The Effect Of T Cell Function On Ovarian Cancer And Its Mechanism

Objective:The purpose of this study is to investigate whether the activation of p38 MAPK signaling pathway can be regulated by changing the expression of Sema4 C in T cells in vitro,and explore whether it can affect cellular biological characteristic of cocultured ovarian cancer cell.Eventually,find the new mechanism for ovarian cancer how to modify immune microenvironment,and preliminarily clarify the mechanism of Sema4 C in T cell immunity of ovarian cancer,so as to lay the foundation for finding new therapeutic targets of ovarian cancer immunotherapy.Methods:1.Structure a lentivirus overexpression carrier of Sema4 C and a lentivirus interference carrier of RNAi lentiviral to transfecte T cell line Jurkat.Then,use Fluorescence quantitative PCR to verify Sema4 C gene’s overexpression and silencing effect,which is transfected by lentivirus in Jurkat cells.2.After the p38 MAPK pathway was blocked by SB203580,a p38 MAPK pathway inhibitor,the expression of p38 MAPK signaling pathway related molecules in Jurkat cells was detected by fluorescence quantitative PCR and Western blot.3.Use CCK-8,colony formation assay and flow cytometry technology to vetify the influence of Sema4 C to Jurkat cells,which is about the proliferation and apoptosis of Jurkat cells.Use ELISA to test the expression of IL-4,IL-10,IL-13,IFN-γ and TNF-α.4.SKOV3 cells were co cultured with lentivirus transfected Jurkat cells to establish SKOV3 / Jurkat co culture system.And then use CCK-8,clone formation assay,Transwell assay and scratch healing assay technology to terrify the chenge of SKOV3 cells’ s proliferation,apoptosis,invasion and migration.Western blot was used to detect the expression of EMT related proteins in SKOV3 cells.Results:1.Jurkat-op-sema4 c cells and jurkat-si-sema4 c cells were successfully constructed.2.The overexpression of Sema4 C in vitro enhanced the proliferation of Jurkat cells,reduced the level of apoptosis,promoted the expression of cytokines ifn-γ,tnf-α,IL-13,and inhibited the expression of cytokines IL-4 and IL-10.On the contrary,Jurkat cells with Sema4 C knockdown had opposite results.3.Overexpression of Sema4 C enhanced the proliferation of Jurkat cells by activating p38 MAPK signaling pathway,which was inhibited by the addition of pathway inhibitors.4.The Jurkat cells overexpressing Sema4 C promoted the proliferation,migration,invasion and epithelial mesenchymal transition of ovarian cancer cells.By adding pathway inhibitors or replacing them with Sema4 C knockdown Jurkat cells,these biological characteristics were effectively reversed.Conclusions:Sema4C can promote the proliferation and cytokine secretion of Jurkat cells by activating p38 MAPK signaling pathway.In the co culture system of T cells and ovarian cancer cells,the expression of Sema4 C in Jurkat cells was regulated,which significantly affected the proliferation,migration,invasion and epithelial mesenchymal transformation of SKOV3.