Effects OTUD7B On The Migration,Proliferation,Apoptosis Of Breast Cancer Cells And The Expression Of Vascular Endothelial Growth Factor

Objective:In the world,the threat of breast cancer to the damage of women has been not optimistic.The occurrence and development of breast cancer are the result of the interaction of multiple genes and pathways and reactions.Which the incidence of the disease and case fatality rate are increasing year by year.The rate of tumor proliferation,metastasis and metastasis sites have a direct impact on the survival of breast cancer patients.In recent years,the age of onset of breast cancer patients tend to be younger,the incidence of invasion and metastasis are greatly increased,as the result many patients have lost the opportunity of surgery,which directly affect the overall survival of the patients.Therefore,inhibiting the growth and blocking the metastasis of tumor play an important role in improving the survival rate and the quality of life of the patients.However,tumor angiogenesis is the fundamental source to lead to malignant tumor development and metastasis,so inhibit breast cancer cell metastasis is important significance for the treatment of breast cancer.Nuclear factor-Kappa B(NF-kappa B)can regulate the expression of vascular endothelial growth factor(VEGF),and the regulation of NF-kappa B is very important for the regulation of deubiquitinating enzymes.It has been found that NF-kappa B can not only promote the expression of VEGF in gastric cancer,thyroid cancer,breast cancer,but also promote lymph node metastasis.OTUD7 B can inhibit the activation of NF-kappa B,while NF-kappa B can regulate the migration and invasion of cancer cells and the expression of vascular endothelial growth factor.Therefore,it is possible to provide a new method for the treatment of breast cancer by proving the relationship between OTUD7 B and proliferation,migration,apoptosis and VEGF expression of breast cancer cells.Methods: To construct lentiviral RNA interference silencing vector(LV3-si-hOTUD7B)and control vector(LV3-NC).Construction of human OTUD7 expression plasmid(pEGFP-h OTUD7B)and control plasmid(pEGFP-CI)with green fluorescent protein tags.Being infection to MCF-7 cells with LV3-si-hOTUD7B,LV3-NC,pEGFP-hOTUD7B and pEGFP-CI lentivirus.MTS shows that effect of OTUD7 B on proliferation of MCF-7cells.Cell scratch test is used to analyze the effect of OTUD7 B on the migration of MCF-7 cells.Flow cytometry PI staining is used to detect the effect of OTUD7 B on cell cycle.Western blot was used to analyze the level of expression of cytoplasmic NF-kappa B P65 and nuclear NF-kappa B P65 after detection of OTUD7 B on MCF-7 cell.Effect of OTUD7 B on the relative expression level of VEGF m RNA by RT-PCR.ELISA shows that OTUD7 B regulates VEGF secretion in MCF-7 breast cancer cells.SPSS19.0 statistical software for processing,all the data to mean ±standard deviation(?).Mean comparisons using t test or one-way ANOVA,with P < 0.05 as a statistically significant difference.Results: The results of MTS assay show that the proliferation of MCF-7 breast cancer cells is inhibited after transfection with pEGFP-hOTUD7B,and the proliferation of MCF-7 breast cancer cells is enhanced after transfection with LV3-si-hOTUD7B.Cell scratch test shows that after transfection with pEGFP-hOTUD7B,the migration of MCF-7 breast cancer cells is inhibited.The migration ability of MCF-7 breast cancer cells is increased after transfection with LV3-si-hOTUD7B.Flow cytometry FCM PI staining results indicate that pEGFP-hOTUD7B can regulate the cell cycle of MCF-7 cell cycle arrest in G1 phase.With the negative control group and normal control group comparison,western blot analyses that pEGFP-hOTUD7B can down-regulated the expression of MCF-7 in breast cancer cell nuclear protein NF-kappa B P65,and cytosolic NF-kappa B P65 has no significant difference.RT-PCRresults suggest that PEGFP-hOTUD7B inhibits the expression of VEGF.ELISA shows that compared with the negative control group and the normal control group,the VEGF content in the conditioned medium of the pEGFP-hOTUD7B group was lower.Conclusion: OTUD7 B can inhibit the proliferation and migration of MCF-7 breast cancer cells,which regulate the MCF-7 cell cycle arrest in G1 phase,as the same time,inhibition of MCF-7 breast cancer cells express and secrete VEGF.