GPR35 Gene Regulates PI3K/AKt/mTOR Signal Pathway To Proliferate Breast Cancer Cells

Objective: To investigate molecularly the effect of G protein-coupled receptor 35(G Protein-Coupled Receptor 35,GPR35)regulating phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway on breast cancer cell proliferation,and the upregulation of GPR35 gene causing tumor necrosis The study of the expression changes of factor receptor related protein 1 on the pathogenesis of breast cancer.Methods:Breast cancer(MDA-MB-231)cells were cultured first,genetic modification methods were used to construct a model of overexpressing GPR35 cells used in the research,Western blot and immunofluorescence were used to detect the expression of GPR35 in breast cancer cells;MTT method was used to detect overexpressing GPR35 Changes in the proliferation rate of breast cancer cells afterwards.q PCR method was used to detect the expression levels of PI3 K,AKt1,AKt3,mTOR,and TRAP1 mRNA in breast cancer cells after overexpression of GPR35;Western blot was used to verify gene expression,and to detect the expression of regulatory signaling pathway proteins PI3 K,total AKt,total mTOR and TRAP1 protein The expression of p-PI3 K,p-AKt,and p-mTOR protein in breast cancer cells after overexpression of GPR35 was further detected.Results: Western blot method was used to detect the expression level of GPR35 protein in breast cancer cells overexpression GPR35 a group and overexpression GPR35 b group was higher than that in normal group(p<0.05),and the GPR35 protein expression level in the overexpression GPR35 b group was higher than that in the overexpression GPR35 a group(p<0.05);Cellular immunofluorescence staining experiments determined that GPR35 was mainly located in the cytoplasm.Compared with the normal group,the expression of GPR35 protein(green)in breast cancer cells of the overexpression GPR35 a group and the overexpression GPR35 b group was significantly increased;MTT method was used to detect breast cancer Cell proliferation experiments showed that the proliferation rate of breast cancer cells in the overexpression GPR35 a group and the overexpression GPR35 b group was significantly higher than that of the normal group(p<0.05),and the proliferation rate of breast cancer cells in the overexpression GPR35 b group was higher than that in the overexpression GPR35 a group(p<0.05);q PCR experiment detected the expression levels of PI3 K,AKt1,AKt3 and mTOR genes in breast cancer cells.There was no significant change in the pairwise comparison among the normal group,the overexpression GPR35 a group and the overexpression GPR35 b group(p>0.05),The expression level of TRAP1 mRNA in breast cancer cells was detected by q PCR experiment.The expression level of TRAP1 mRNA in the overexpression GPR35 a group and the overexpression GPR35 b group was higher than that in the normal group(p<0.05),and the expression level of TRAP1 mRNA in the overexpression GPR35 a group was higher In the overexpression GPR35 b group(p<0.05);Western blot method was used to detect the expression levels of PI3 K,total AKt,and total mTOR protein in breast cancer cells.The normal group,the overexpression GPR35 a group and the overexpression GPR35 b group were compared in pairs.There was no significant change(p>0.05),which was consistent with the gene expression level;The expression levels of p-PI3 K,p-AKt,p-mTOR,and TRAP1 protein in the overexpression GPR35 a group and the overexpression GPR35 b group were higher than those in the normal group(p<0.05),and the expression levels of p-PI3 K,p-AKt、p-mTOR and TRAP1 protein in the overexpression GPR35 b group were higher than those in the overexpression GPR35 a group(p<0.05).Conclusions:(1)GPR35overexpression can promote the proliferation of breast cancer cells(MDA-MB-231),which is closely related to the occurrence and development of breast cancer.(2)GPR35 activates the PI3K/AKt/mTOR signaling pathway through phosphorylation,up-regulates TRAP1 mRNA and protein expression,and promotes the proliferation of breast cancer cells.