The Role Of C1QBP In Renal Carcinoma Progression

Objectives:Renal carcinoma is one of the most common urological cancer,with high incidence in China that seriously threaten human health.Tumor development is closely related to cell proliferation,chemotaxis,adhesion and invasion ability.There are many unknown fields to be explorated on targeted therapy of renal carcinoma.Therefore,in-depth study on molecular mechanism of renal carcinoma development and discovering new biomarker have become urgent problems to be solved. C1QBP is highly expressed in a variety of malignant tumors in the previous studies,At the same time,C1QBP is also reported low expressed in cervical carcinoma,which shows that the expression and regulation of tumor biological behavior of C1QBP have tissue specificity,but relevant research of C1QBP in renal carcinoma has not been reported so far.Therefore,in this study we will explore C1QBP expression in renal carcinoma and the influence of C1QBP on cell proliferation,chemotaxis,adhesion and invasion ability during the development of renal carcinoma,initially identify the role of C1QBP in renal carcinoma development. Methods:Western Blotting,Realtime PCR and immunohistochemical analysis were used to detect the expression of C1QBP in renal carcinoma and tissue adjacent to carcinoma;statistically analyzed the correlation between C1QBP expression and clinicopathological factors.Realtime PCR was performed to detect C1QBP expression in renal epithelial cell and renal carcinoma cell lines.C1QBP knockdown stable renal carcinoma 786-0 cell line was constructed by lentivirus mediated RNAi technology,Western Blotting and Realtime PCR were used to verify its expression.CCK8 assay,chemotaxis assay,matrigel invasion assay and adhesion assay were respectively utilized to examine the effect of C1QBP knockdown on proliferation ability,EGF induced migration ability,cell invasion ability and EGF induced adhesion ability of renal carcinoma cell 786-0.Through genchip detection,the influence of C1QBP knockdown on gene expression was analysed in renal carcinoma cell,and differentially expressed genes were verified by Realtime PCR.Western Blotting was used to examine the molecular mechanism in renal carcinoma process regulated by C1QBP.SCID mice were adopted to investigate the effect of C1QBP knockdown on proliferation and metastasis of renal carcinoma transplanted by 786-0 cell line,and tested the expression of proliferation related antigen PCNA by immunohistochemical. Results:1. Compared with para-carcinoma tissue,RNA and protein expression of C1QBP were lower in renal carcinoma tissue,and C1QBP expression was significantly associated with clinical histological staging and survival rate after operation.2. C1QBP was lower expressed in renal cacinoma cells than in the renal epithelial cell,and C1QBP expression gradually reduced with the increasing malignance of renal cacinoma cells.3. Lentivirus mediated RNA interference technology had been successfully applied to establish C1QBP knowdown stable cell line 786-0,Western Blotting and Realtime PCR comfirmed that protein and gene expression of C1QBP were significantly lower.4. The proliferation ability,EGF induced migration,cell invasion and EGF induced adhesion ability of renal carcinoma cell 786-0 were obviously increased than the control group cell after C1QBP knockdown(P< 0.05).5. Through gene chip analysis,C1QBP knockdown could affect the gene expression related to cell adhesion,eight adhesion related genes were further tested by Realtime PCR,L1 CAM was significantly higher in 786-0-shC1QBP among these genes.6. After C1QBP knockdown,the expression of ?-catenin in renal carcinoma cell was increased,likewise the expression of p-GSK3 was also increased.7.C1QBP knockdown promoted renal carcinoma transplanted tumor growth and metastasis in vivo,the PCNA expression was obviously higher than the control group. Conclusions:1. Compared with the para-carcinoma tissue,C1QBP expression was lower in renal carcinoma tissue,and the expression of C1QBP was associated with histological staging and survival rate of patients,furthermore C1QBP expression was gradually reduced with the increasing malignance of renal cacinoma cells.2. Knocking down C1QBP could promote cell proliferation,migration and invasion ability of renal carcinoma.3. C1QBP knockdown could promote cell adhesion ability of renal cacinoma cell,possibly by activating Wnt pathway related proteins,and play a biological role by up regulating LICAM.4. C1QBP knockdown might promote proliferation and metastasis ability of renal carcinoma cell in vivo,indicating that C1QBP is a key factor in regulating renal carcinoma development.C1QBP could be not only a novel marker for renal carcinoma diagnosis and prognosis,but also promising to be a new molecular target for the treatment of renal carcinoma.