Effects Of Silencing TRAF4 Gene On Biological Behaviors In Human Breast Cancer Cells And Explore Its Mechanism

Objective: To investigate the effect of proliferation,migration,invasion and apoptosis in the breast cancer cells MDA-MB-231 after silencing TRAF4 gene,and its possible mechanism.And to provide theoretical evidence for new targeted gene therapy of breast cancer.Methods:1.Lentivirus vector was used to infect MDA-MB-231 cells,and the expression of TRAF4 mRNA and protein in silenced MDA-MB-231 cells was detected by real-time fluorescence quantitative PCR and Western blotting.We can verify the effect of silence and screen for the best target for silence.MDA-MB-231 cells were blank control group.MDA-MB-231 cells transfected with LV-shRNA-NC were negative control group,and MDA-MB-231 cells transfected with LV-shRNA-TRAF4 were experimental group.2.The effect of silencing TRAF4 gene on the proliferation of MDA-MB-231 cells was detected by CCK-8 cell proliferation assay and plate colony formation assay.Transwell migration assay and wound-healing assay were used to detect the effect of silencing TRAF4 gene on migration of MDA-MB-231 cells.Transwell invasion assay was used to detect the effect of silenced TRAF4 gene on invasion of MDA-MB-231 cells.Flow cytometry wasused to detect the apoptosis of MDA-MB-231 cells before and after silence of TRAF4 gene.3.The expression levels of PRMT5 and NF-?B were detected by Western blotting before and after TRAF4 gene silencing,to explore the possible mechanism of the effect of silencing TRAF4 gene on the biological function of human breast cancer cell line MDA-MB-231.Results:1.Successfully constructed a human breast cancer cell line MDA-MB-231 that silences the TRAF4 gene.Real-time fluorescence quantitative PCR and Western blotting were used to detect the effect of silencing.The results showed that the expression level of TRAF4 mRNA and protein in the experimental group was significantly lower than that of the blank control group and the negative control group(P<0.05).2.CCK-8 proliferation assay and plate colony formation assay results showed that the silence of TRAF4 gene in human breast cancer cell line MDA-MB-231,the proliferation of the experimental group was significantly lower than that of the blank control group and the negative control group.(P<0.05).3.The results of wound-healing test and cell migration experiment(Transwell)showed that silencing TRAF4 gene in human breast cancer cell line MDA-MB-231,the cell migration of the experimental group was lower than that of the blank control group and the negative control group(P<0.05).4.The results of Transwell assay showed that silencing of TRAF4 gene in human breast cancer cell line MDA-MB-231,the cell invasion ability of the experimental group is lower than that of the blank control group and the negative control group(P<0.05).5.Flow cytometry analysis of cell apoptosis showed that silencing TRAF4 gene in human breast cancer cell line MDA-MB-231,the apoptosis rate of the experimental group was higher than that of the blank control group and the negative control group(P<0.05).6.After silencing the TRAF4 gene in human breast cancer cell line MDA-MB-231,the expression of PRMT5 and NF-?B decreased.Conclusion:1.LV-shRNA-TRAF4 can stably transfect human breast cancer cells MDA-MB-231.2.Silencing TRAF4 gene can inhibit the proliferation,migration and invasion of human breast cancer cell line MDA-MB-231 and induce apoptosis.3.The effect of silent TRAF4 gene on the biological function of human breast cancer cell line MDA-MB-231 may be related to PRMT5 and NF-?B pathway.