High Expression Of KIF20A Promotes The Development Of Clear Cell Renal Cell Carcinoma And Affects The Clinical Prognosis

Objective: To explore the potential biological role and mechanism of KIF20 A in the occurrence and development of renal clear cell carcinoma.Method: TCGA database and bioinformatics technology were used to screen the key genes related to the occurrence and development of renal clear cell carcinoma,and GEPIA online tool was used for survival analysis.Reverse transcriptase polymerase chain reaction(RT-PCR)and Westernblot were used to detect the expression of KIF20 A gene at m RNA level and protein level in10 cases of clear cell renal cell carcinoma(cc RCC)and their matched paracancerous tissues.The cell culture included four cc RCC cell lines ACHN,786 Mel O,Caki-1 and Caki-2 and a human renal tubular epithelial cell line HK2.The cell lines with relatively high expression of KIF20 A were identified by Western blot.Lentivirus was used to infect cell lines with three different sh RNA sequences packaged,the infection gradient was set according to the MOI value of the cell line.The efficiency of KIF20 A silencing was judged by the intensity and ratio of GFP fluorescence,and the stable transformants were screened with puromycin.The proliferation of tumor cells was detected by CCK8,the apoptosis and cell cycle of tumor cells were detected by flow cytometry,the migration and invasion ability of tumor cells were detected by wound healing assay and Transwell,the effect of KIF20 A expression level on tumor growth was detected by nude mouse xenograft model.Result: The results of GO and KEGG analysis showed that the differentially expressed genes of cc RCC in TCGA database were mainly involved in 10 biological processes,such as inflammatory response,cell surface receptor signal pathway,secretion and so on.The main enrichment pathways of KEGG are cell adhesion molecules,cytotoxicity mediated by natural killer cells,rejection,cytokine-cytokine receptor interaction and so on.Finally,seven core candidate genes were screened: ASPM,NCAPG,NUSAP1,KIF20 A,Bub1,AURKB and TPX2.The results of gene differential expression analysis showed that the expression of Bub1 and KIF20 A in tumor tissues was higher than that in paracancerous tissues(P < 0.001).Survival analysis showed that patients with high expression of ASPM,AURKB,Bub1,TPX2 and KIF20 A genes had lower overall survival(OS)and disease-free survival(DFS).The results of PCR and Westernblot experiments showed that the expression of KIF20 A in renal clear cell carcinoma was higher than that in paracancerous tissues at both protein level and m RNA level(P < 0.001).The results of immunohistochemistry in 72 cases of renal clear cell carcinoma showed that the patients with high expression of KIF20 A had higher tumor grade(P =0.004),later T stage(P =0.017)and larger tumor volume(P =0.040).ACHN and 786-o with relatively high expression of KIF20 A were selected for cell experiment in vitro.The results of CCK-8 assay showed that KIF20 A gene silencing could inhibit the proliferation of ACHN and 786 Mel O cells,while Transwell assay showed that KIF20 A gene silencing inhibited the invasion and migration of renal clear cell carcinoma.Flow cytometry showed that KIF20 A gene silencing could block the cell cycle progression of tumor cells in S/G2 phase and induce tumor cell apoptosis.The results of subcutaneous tumorigenesis in nude mice showed that the tumor derived from KIF20 A gene silencing cc RCC cells was smaller.Conclusion: the expression of KIF20 A is up-regulated in cc RCC,and its high expression indicates poor clinical prognosis.Silencing KIF20 A can inhibit the proliferation,migration and invasion of tumor cells.It is suggested that KIF20 A may be a potential biomarker and therapeutic target for patients with cc RCC.